{"paper_id":"2df81f1d-fa9d-439a-9d82-e2e0d1fbe44e","body_text":"Quercetin loaded and alginate sealed β-Glucan particles-based drug delivery system against DU145 a prostate cancer cell line: Integrating network pharmacology, molecular docking and in vitro studies | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Quercetin loaded and alginate sealed β-Glucan particles-based drug delivery system against DU145 a prostate cancer cell line: Integrating network pharmacology, molecular docking and in vitro studies RASHMI TRIVEDI, Tarun Kumar Upadhyay, Pranav Kumar Prabhakar This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4486275/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Background Prostate cancer remains a challenge in healthcare, being the second most common male cancer demanding innovative therapeutic approaches and treatment techniques. This study integrates in silico and in vitro methods for the investigation of the potential anticancer effects of quercetin-loaded and alginate-sealed β-Glucan particles derived from mushroom Agaricus bisporus and yeast against the DU145 cell line. Methods Prostate cancer-related genes were identified from DisGeNET and GeneCards databases, followed by target prioritization using Swiss Target Prediction software. Venny 2.1 was used for the determination of common targets between β-Glucan, Quercetin, and prostate cancer and PPI network was constructed using STRING database. CB dock online server was used for molecular docking and DU145, RAW264.7 cell lines were used for the determination of cytotoxicity against prostate cancer and healthy cells. Results Molecular docking revealed that quercetin has superior binding affinity compared to β-Glucan with selected prostate cancer-related targets. In vitro evaluation using MTT assays demonstrated the cytotoxic effects of quercetin-loaded and alginate-sealed particles against DU145 prostate cancer cells. Apoptosis induction, ROS generation, and lysosomal pH alterations underscore the potential of quercetin-loaded and alginate-sealed β-Glucan particles as promising therapeutic agents for prostate cancer. Conclusions Our study showed systematic analyses of the effect of hollow β-Glucan particles, Quercetin, and Quercetin alginate sealed particles against DU145 cells and found that formulation exhibits superior anticancer activity against prostate cancer cell line. Quercetin-loaded alginate-sealed particles showed very little cytotoxicity against healthy cell line RAW264.7. Future studies focusing on preclinical validation, pharmacokinetic profiling, and clinical trials to assess translational potential and optimize therapeutic strategies can help get impactful findings. β-Glucan DU145 prostate cancer Quercetin network pharmacology Apoptosis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Background Prostate cancer is a tumor that occurs in the prostate gland due to uncontrolled growth of the cells and represents a huge challenge in the landscape of oncology. When cells of prostate cancer start to metastasize to different organs, the survival rate of the patient starts to decline significantly (Atasoy and Erbaş 2020 ). The process of metastasis and the tendency of prostate cancer cells to target specific organs are influenced by various cellular subtypes and the unique microenvironments of those organs, along with the interactions between them (Klusa et al., 2021 ). The androgen hormone, testosterone, and its metabolite dihydrotestosterone (DHT) play pivotal roles in prostate cancer development and progression. Binding of androgen to the androgen receptor leads to the activation of cell proliferation signaling leading to the development of prostate cancer. Androgen receptor signaling pathways lead to the proliferation and survival of prostate cells in the malignant stage, making androgen deprivation therapy (ADT) through the removal of gonads a cornerstone in the management of advanced disease (Hou et al., 2021 ). Gonadotropin-releasing hormone (GnRH) is found to be involved in the inhibition of steroidogenesis leading to the suppression of androgen receptors resulting in good management of prostate cancer (Oduwole et al., 2021 ). Symptoms of prostate cancer are nonspecific or may remain asymptomatic until later stages. Screening strategies, including serum prostate-specific antigen (PSA) and digital rectal examination (DRE) measurement, are the techniques that are used for the early-stage detection of prostate cancer (Morote et al., 2022 ). Currently available treatment protocols for prostate cancer, including surgical interventions, radiation therapy, hormone therapy, chemotherapy, and immunotherapy, offer valuable options for patients still, each treatment comes with its set of limitations and potential side effects that can significantly impact well-being and quality of life of the patient that leads to the continued research in prostate cancer treatment (Board 2023 ). Surgical interventions, such as radical prostatectomy, are effective in removing localized tumors, however, they can lead to complications such as urinary incontinence, erectile dysfunction, and bowel dysfunction. Polysaccharides are a digestible and indispensable component present in a variety of foods that provide essential calories and glucose necessary for regular bodily functions along with various health benefits. One extensively studied and well-documented bioactive polysaccharide is β-Glucan having monomer units linked through glycosidic bonds at β 1→3, 1→4, and/or 1→6 positions, forming branched or unbranched structures (Kaur et al., 2019 ). β-Glucans are carbohydrate polymers and they are the main constituents of the cell walls of algae, lichens, yeasts, fungi, and bacteria, as well as in many plants and have a molecular weight in the range between 50 kDa to 4100 kDa contributing to their diverse biological activities (Nakajima et al., 2024 ). β-Glucans exhibit a rigid structure characterized by β-1→3 and 1→6) glycosidic linkage in the case of yeast and mushrooms, whereas β-1→3 and 1→4) glycosidic linkages in cereals. The molecular weight of β-Glucans is influenced by various factors, including the type of detectors utilized, reaction conditions, solvents employed, and the specific compounds present in the sample. These parameters collectively determine the size and distribution of β-Glucan molecules, impacting their functional properties and physiological effects (Kupetz et al., 2015 ). β-Glucans exhibit diverse biological activities influenced by several key parameters. These include molecular weight, structural characteristics, conformational arrangement such as single helix, triple helix, or random coil, charge on the polymer, and density of branching. Changes or modifications in the conformation of β-Glucans can profoundly impact their immune-modulatory properties and overall functionality (Caseiro et al., 2022 ). Despite being insoluble in water, β-Glucans find extensive applications in the food and pharmaceutical industries. They serve as valuable raw materials for various beverages and medicinal products, contributing to the therapeutics for numerous diseases including cholesterol management, diabetes, and cancer (Sletmoen and Stokke 2008 ). Quercetin is a flavonoid in fruits and veggies, having a ketocarbonyl group and four active hydroxyl groups with scientific name 3, 3', 4', 5, 7-pentahydroxyflavone. The phenolic hydroxyl group and double bond presence make it a strong antioxidant (Yang et al., 2020 ). It boosts anti-inflammatory, antioxidant, and anticancer properties along with offering vasodilatory, anti-hypertensive, anti-hypercholesterolemic, anti-obesity, and anti-atherosclerotic effects. Its standout quality is triggering tumor cell death (apoptosis), hindering the progression of various human cancers like prostate cancer while sparing normal cells. Specifically with prostate cancer, quercetin indirectly hampers key genes like the prostate-specific antigen (PSA), and androgen receptor (AR) potentially impeding cancer development (Fard et al., 2021). Although quercetin is reported to have various biological activities, its lower solubility is one of its drawbacks in medicine so many researchers tried to load it in the nano/microparticles for its targeted delivery and found positive more effective, and improved biological activity (Nazemi et al., 2023 ). Quercetin has the potential for the treatment of cancer but it has some limitations such as low aqueous solubility, getting clear very rapidly from the body, high metabolic rate, and very poor absorption that make it restricted for use in cancer therapy. To overcome these limitations, the use of a delivery agent/carrier may enhance its absorption, solubility, and target specificity (Nam et al., 2016 ). In a variety of cancers including prostate cancer, Quercetin is found to inhibit a variety of enzymes that are responsible for the activation of carcinogens and cell proliferation signaling pathways. In this study, we have loaded Quercetin in the hollow β-Glucan particles and sealed it with alginate for the slow and sustained release of the Quercetin for the possible treatment of prostate cancer. The novelty of the study lies in the innovative approach of encapsulating quercetin within a β-Glucan carrier and then sealing it with sodium alginate for targeted therapy against prostate cancer. This method addresses several limitations associated with the use of quercetin in cancer therapy, including its low aqueous solubility, rapid clearance, high metabolic rate, and poor absorption. Methods Reagents and chemicals Quercetin (HiMedia), MTT, DAPI, PI, H2DCFDA, LysoTracker Red DND 99, MitoTracker Red CMX-ROS, Acridine orange, Ethidium bromide, Annexin V/FITC were purchased from Invitrogen (Thermo Fisher Scientific). DMEM (HiMedia), fetal bovine serum (FBS), antibiotic, and antimycotic solution, were purchased from Gibco™ . Maintenance of the Cell culture DU145 cell line for the prostate cancer was obtained from the National Centre for Cell Science (NCCS) Pune, India. Cells were maintained at 37°C and 5% CO2 in a CO 2 incubator. Complete DMEM media was supplemented with 10% FBS and 1% antibiotic and antimycotic solution. In silico studies Prostate cancer related gene identification Target genes related to prostate cancer were identified by exploring the term prostate cancer in the GeneCards ( https://www.genecards.org/ ) and DisGeNET ( https://www.disgenet.org/ ) databases and different targets were downloaded into Excel format (Cao et al., 2022 ). The targets from two databases pooled and duplicates were removed to obtain the final targets for the prostate cancer. Collection of drug targets We have selected common targets between β-Glucan and Quercetin using Swiss target prediction software. Common targets related to β-Glucan, Quercetin, and prostate cancer were recognized using Venny 2.1 ( https://bioinfogp.cnb.csic.es/tools/venny/ ). PPI Network construction Protein-Protein Interaction (PPI) network was analyzed using STRING 11.0 database ( https://string-db.org/ ) (Suresh et al., 2021 ). These common potential target genes were merged into the STRING 11.0 database to create a PPI network, species keeping to Homo sapiens. Molecular docking Ligand β-Glucan and Quercetin were imported from PubChem databases. Macromolecules were downloaded in PDB format from RCSB PDB ( https://www.rcsb.org/ ). Molecular docking was performed using CB Dock2 online server ( https://cadd.labshare.cn/cb-dock2/index.php ) (Yang et al., 2020 ). In vitro anticancer activity determination Particle preparation and characterization β-Glucan particles were prepared using the acid-base extraction method from the Agaricus bisporus and yeast and characterization was performed using FTIR analysis. We have already published the preparation and characterization methodologies (Upadhyay et al., 2022 ; Bhosale et al., 2022 ). Loading of quercetin into β-Glucan and alginate sealing was performed according to the previously published protocol (Upadhyay et al., 2017 ). Assessment of the cell viability of DU145 and RAW264.7 cells with MTT assay We have assessed the cell viability using the previously described protocol (Vodnik et al., 2021 ). 10,000 cells DU145 and RAW264.7 (healthy cell line) were placed in each well of a 96-well plate and left to grow for 24 hours in a humid environment with 5% CO2 at 37°C. Following this, the cells were exposed to particles prepared in concentrations of IC50 and below and above IC50 for another 24 hours. After treatment, the old media was removed, and MTT dye (10 µl) was added to each well, followed by an incubation of 4 h at a temperature of 37°C. Next, 100 µl DMSO was added to solubilize the formazan crystals formed by viable cells, and after 10 minutes, the absorbance of the dissolved crystals was computed at 490 nm. Cell viability was then calculated using a provided formula $$\\text{%}\\text{V}\\text{i}\\text{a}\\text{b}\\text{i}\\text{l}\\text{i}\\text{t}\\text{y}=\\left(\\frac{\\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e} \\text{o}\\text{f} \\text{t}\\text{r}\\text{e}\\text{a}\\text{t}\\text{m}\\text{e}\\text{n}\\text{t}}{\\text{A}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e} \\text{o}\\text{f} \\text{c}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l}}\\right)\\text{X}100$$ Concentrations of IC 50 and below and above IC 50 were selected for the further analysis. Morphological observations Morphological changes in the treated cells were observed as per the previously defined protocol (Khan et al., 2020 ). In brief, 50,000 cells/well were added in a 24-well plate and left overnight for the attachment. After 24 h, cells were given treatment with the selected concentrations of the particles and Quercetin and further left for incubation for 24 h at 37oC. After incubation, cells were washed with the PBS, and morphological changes were observed and captured with the help of an EVOS FLoid imaging station. Assessment of Reactive oxygen species (ROS) generation The assessment of reactive oxygen species (ROS) production after particle treatment was conducted according to the protocol described by (Zhu et al., 2022 ). Initially, a cell density of 50,000 cells per well was seeded into a 24-well plate and permitted to incubate for 24 hours to ensure attachment. Following this, the cells were treated with selected concentrations of β-Glucan and Quercetin and then maintained in culture for an additional 24 hours at 37°C with 5% CO2. After the incubation period, the cells were rinsed through phosphate-buffered saline (PBS) and stained with 20µM DCFDA dye, followed by incubation at 37°C for 20 minutes. Finally, images were captured using an EVOS FLoid imaging station. Determination of nuclear morphology and DNA fragmentation with DAPI staining DAPI staining was carried out based on a previously described protocol with slight adjustments, as outlined in the work by (Sana et al., 2020 ). In brief, 50,000 cells/ well were seeded in a 24-well plate and allowed to incubate for 24 h. Following this, the cells were exposed to selected concentrations of the particles and further incubated for 24 h, at temperature 37°C. Once the incubation period was completed, the cells were rinsed with PBS and the cells were stained with 1µg/mL of DAPI for 15 minutes. Morphological changes were then observed and documented. Apoptosis detection with Propidium iodide (PI) staining Cells undergoing apoptosis were determined using PI staining with the help of an already established protocol with minor modification (Yang et al., 2022 ). In Brief, 50,000 cells per well were seeded in a 24-well plate and incubated overnight. After that, cells were treated with a selected concentration of Quercetin and prepared particles and further incubated for 24 h. After incubation, cells were stained with 1µg/ml of the PI (prepared from 1mg/ml of PI) and further incubated for 10 min at 37oC following which images were obtained with the EVOS FLoid imaging station. Acidic organelles activity analysis through LysoTracker Red DND-99 Acidic organelles were detected by using LysoTracker Red (100nM) as per the formerly described method (Ning et al., 2020 ). In brief, 50,000 cells/well seeded in 24 well plate and incubated overnight. After the attachment of the cells, treatment with the various concentrations of particles was performed and further incubated for 24 h. Post incubation, cells were washed with PBS and stained with LysoTracker for 30 min and imaged under a FLoid imaging microscope. Mitochondrial membrane potential (MMP) assessment MMP was assessed as per the previously reported protocol (Fernandes et al., 2013 ). Briefly, 50,000 cells/well were seeded in 24 well plate and incubated for 24 h, afterwards, cells were treated with the concentrations of particles and further incubated for 24 hrs. After incubation, cells were stained for 30 min using the dye MitoTracker red CMX ROS (300nM) and imaged under EVOS FLoid imaging microscope. Acridine orange and Ethidium bromide (AO/EtBr) dual staining AO/EtBr dual staining was performed conferring to the earlier described protocol with slight modification (Bobadilla et al., 2019 ). 50,000 cells/ well seeded in 24 well plate and after gaining confluency, treated with selected concentrations of particles and left for 24 h incubation in a CO2 incubator. After that, cells were stained with 5 µl of AO and 5 µl EtBr (5mg/ml each), and cell death was observed with a FLoid Imaging microscope. To check anticancer activity via Wound healing/Cells Scratch assay To determine the scratch healing ability, DU145 cells were seeded at a density of 50000 cells/ well in a 24-well plate and left overnight for adhesion. Before treatment, a scratch was made with the help of a sterile 10µl tip and cells were imaged followed by treatment with different doses of the particles. After 24 h, cells were imaged again to quantify the change in scratch area under an inverted microscope. The area of the scratch or wound was quantified by the given formulae (Yusof et al., 2019). $$\\text{W}\\text{o}\\text{u}\\text{n}\\text{d} \\text{a}\\text{r}\\text{e}\\text{a} \\text{p}\\text{e}\\text{r}\\text{c}\\text{e}\\text{n}\\text{t}=\\frac{\\text{A}\\text{t}}{\\text{A}0}\\text{x}100$$ Where A t = wound area after treatment and A 0 = Wound area before treatment. AnnexinV/FITC and Propidium iodide dual staining Apoptotic cells were analyzed using AnnexinV/FITC and PI dual staining as per the previously reported protocol. Briefly, DU145 cells were seeded at a density of 50000 cells/well in a 24-well plate and left overnight for the incubation. After the attachment of the cells, they were treated with IC50 concentrations of the prepared particles and left for another 24 h. After incubation, cells were washed through PBS and stained with 1µl of AnnexinV/FITC and PI each and images under a fluorescence microscope (Lin et al., 2020 ). Statistical analysis In vitro experiments were accomplished in triplicates and One-way ANOVA was used for the statistical analysis using GraphPad Prism 8.0 and ImageJ. A probability value of p < 0.05 was deliberated statistically significant where *** means highly significant, ** means less significant and * means least significant. Results In silico studies Prostate cancer related gene identification 683 gene were retrieved from DisGeNET while 13832 genes were identified from GeneCards. Common genes were retrieved using Venny 2.1 after removing duplicate genes. Collection of drug targets Drug targets were retrieved using Swiss target prediction software. 100 targets were downloaded as comma separated value for both β-Glucan and quercetin. Common targets were retrieved using Venny 2.1 as shown in Fig. 1. Construction of PPI Network PPI network of common genes of β-Glucan, Quercetin and prostate cancer is shown in Fig. 2a. After intersection of common targets of β-Glucan and quercetin with common targets of prostate cancer, we obtained 3 gene as shown in Fig. 2b that we have taken for the molecular docking. Molecular docking Ligand β-Glucan and Quercetin were docked using CB dock2 with CYP19ABG (PDB ID: 5JKW), ABCB1 (PDB ID: 6QEX), and CDK1 (PDB ID: 6TWN). Interaction of ligands and targets is shown in Fig. 3. In each docking, quercetin shows higher binding affinity with the targets than β-Glucan. The binding energies of ligands and targets and their interacting amino acids are presented in Table 1. Table 1 Binding energies and interacting amino acids of the ligand and target docking. S. No. Target Ligand Binding energy Interacting Amino acids 1 ABCB1 Quercetin -8.4 Chain A : TRP232 ILE235 LEU236 SER238 PHE239 THR240 ASP241 LYS242 GLU243 LEU244 ALABG + Q46 TYR247 ALABG + Q84 LYS285 GLYBG + Q88 ILE289 LYS291 ALABG + Q92 ALABG + Q95 ASN296 ILE299 PHE303 PHE770 GLN773 GLY774 PHE777 GLY778 ALA780 GLY781 GLU782 THR785 LYS786 ARG789 ALA823 LYS826 GLY830 SER831 ALA834 VAL835 GLN838 MET876 GLN990 VAL991 PHE994 ALA995 PRO996 β-Glucan -7.9 Chain A : PHE239 THR240 ASP241 LYS242 GLU243 LEU244 ALABG + Q46 TYR247 ALABG + Q84 LYS285 ILE287 GLYBG + Q88 ILE289 LYS291 ALABG + Q92 ALABG + Q95 ASN296 PHE770 GLN773 GLY774 PHE775 PHE777 GLY778 LYS779 ALA780 GLY781 GLU782 ILE783 THR785 LYS786 ARG789 ALA823 LYS826 GLY827 GLY830 SER831 ALA834 VAL835 GLN838 VAL991 PHE994 ALA995 PRO996 2 CDK1 Quercetin -8.0 Chain A : THR1465 ALABG468 ARG1469 GLN1472 Chain B : LEU1367 LEU1370 GLU1371 ARG1374 GLU1375 GLU1378 LEU1607 GLU1608 ALABG610 GLYBG611 GLYBG612 β-Glucan -7.5 Chain A : THR1465 ALABG468 ARG1469 GLN1472 Chain B : LEU1367 LEU1370 GLU1371 THR1372 ARG1374 GLU1375 GLU1378 LYS1604 LEU1607 GLU1608 SER1609 GLYBG611 GLYBG612 GLN1615 3 CYP19ABG Quercetin -7.8 Chain A : ARG115 ILE132 ILE133 PHE134 ARG192 SER199 PHE203 GLN218 PHE221 ASP222 TRP224 GLN225 GLU302 MET303 ILE305 ALABG + Q + ALG06 ALABG + Q + ALG07 PRO308 ASP309 THR310 MET311 SER312 VAL313 SER314 MET364 VAL369 VAL370 LEU372 VAL373 MET374 ARG375 PRO429 PHE430 GLY436 CYS437 ALA438 GLY439 ILE442 ALA443 MET446 MET447 ILE474 LEU477 SER478 LEU479 HIS480 PRO481 ASP482 GLU483 THR484 β-Glucan -7.4 Chain A : ARG115 ILE132 ILE133 PHE134 TRP141 ARG145 PHE148 LEU152 LEU188 ARG192 PHE203 ILE217 GLN218 GLYBG + Q19 PHE221 ASP222 TRP224 GLN225 ALABG + Q26 GLU302 MET303 ALABG + Q + ALG06 ALABG + Q + ALG07 PRO308 ASP309 THR310 MET311 SER312 VAL313 SER314 MET364 VAL369 VAL370 LEU372 VAL373 MET374 ARG375 ILE398 PRO429 PHE430 GLY431 ARG435 GLY436 CYS437 ALA438 GLY439 LYS440 ILE442 ALA443 MET446 MET447 ILE474 LEU477 SER478 HIS480 PRO481 ASP482 GLU483 THR484 Determination of anticancer activity Particle preparation Prepared β-Glucan particles from the Agaricus bisporus and yeast, loaded with quercetin and alginate sealed were used for the determination of in vitro anti-cancer activity as shown in Fig. 4. We hypothesized the slow and sustained release of the quercetin from β-Glucan after alginate sealing as is reported in many studies (Ameer et al., 2024). Hollow β-Glucan from Agaricus bisporus and yeast were named ABG and YBG, Quercetin loaded were named ABG + Q, YBG + Q, and Alginate sealed particles were named ABG + Q + ALG, YBG + Q + ALG respectively. Q is used for quercetin here. Assessment of the cell viability with MTT assay on DU145 and RAW264.7 cells Cell viability of the DU145 cells upon exposure to the particles derived from Agaricus bisporus and yeast was found to decrease as shown in Fig. 5 and Fig. 6. Yeast-derived particles were found to have lower IC50 than the Agaricus bisporus derived particles. In the case of yeast-derived particles, quercetin-loaded-alginate-sealed particles have the lowest IC50. Concentrations of IC50 and below and above IC50 were taken for the further microscopic examination of anticancer activity. To evaluate the consequence of the particles on the healthy cells we treated RAW264.7 cells with the serial dilution from 500µg/ml to 1.56µg/ml. We found that particles did not have cytotoxic effects at initial concentrations as shown in Fig. 7 and at higher concentrations of 500 and 250µg/ml, particles had very little cytotoxic effects on the RAW264.7 cells. Morphological observations With the help of FLoid imaging microscope, we observed clear morphological changes in the shape of the DU145 cells. At the highest concentration, cells were highly disrupted and they lost their actual shape as shown in Fig. 8. Assessment of Reactive oxygen species (ROS) generation ROS are a natural derivative of oxygen metabolism in the body and have a variety of important roles in cell signaling. However, excessive ROS generation can lead to oxidative damage leading to the apoptosis of the cancer cells (Sahoo et al., 2022 ). In this study, we found that β-Glucan and the quercetin loaded β-Glucan particles have the potential to generate an efficient amount of ROS in DU145 cells as shown in Fig. 9. Determination of nuclear morphology and DNA fragmentation with DAPI staining The influence of particles on the DU145 cells was assessed with DAPI staining that is specific for the binding with the nucleus. Nuclear condensation and abnormalities are quite observable and can be seen in Fig. 10. We have found that nuclear deformities were increasing with the increase in the treatment concentration of the particles. Apoptosis detection with Propidium iodide (PI) staining PI is a very specific dye that only binds with the dying or apoptotic cells resulting in red color fluorescence. Cells in the late apoptotic or early necrotic stage stain red. In our study we have found a significant increase in the apoptotic cells with the increase in concentration as shown in Fig. 11. In non-treated (control) cells, there was no significant percentage of apoptosis was observed. LysoTracker Red DND-99 staining to monitor pH change in lysosome Lysosome functioning depends on the lysosomal pH and a decrease in the fluorescence intensity of the red color indicates the elevation in the pH of lysosomes. We have found a dose-dependent decrease in the fluorescence intensity indicating the pH alteration that can result in the apoptosis of the cells as shown in Fig. 12. Mitochondrial membrane potential determination MitoTracker Red CMX-ROS is a probe-based cationic dye that enables to assess the change in MMP. In our study, DU145 cells showed significant decline in MMP after treatment with Agaricus bisporus and yeast derived particles as shown in Fig. 13. Acridine orange and Ethidium bromide (AO/EtBr) staining for the detection of cells in early and late stage of apoptosis AO/EtBr dual staining is considered a very efficient method for the detection of early and late apoptotic cells. AO dye is permeable to all the cells and gives florescence in green color while EtBr is only permeable to the cells undergoing apoptosis and gives red fluorescence. In the dual staining, cells in the early apoptotic stage show a yellowish-orange color while cells in the late apoptotic stage show an orange-red color. We have found cells appeared as yellow-orange-red color cells in early and late apoptosis as shown in Fig. 14. To check anticancer activity via Wound healing/Cells Scratch assay In the wound healing assay cells in the control were able to grow back and cover most of the scratch are up to 55% as shown in Fig. 15. In the cells treated with alginate-sealed particles, the scratch area increased by 10–15% showing that cells can grow and migrate well after the treatment with particles. Annexin V/ FITC and PI dual staining In the Annexin V/ FITC and PI dual staining, it was found that no significant death in the Untreated cells. Cells in the early stages of apoptosis show green fluorescence due to the Annexin V/FITC stain. Cells that are in the later stages of apoptosis or necrotic cells allow PI to enter into the cells due to the membrane disintegration and give red color fluorescence (Kumar et al., 2021 ). In this dual staining method, exposure with dummy β-Glucan particles leads to some of the cells in the apoptotic stage while quercetin-loaded particles are reported to have a significant number of cells in the later stage of apoptosis. Cells that were treated with quercetin loaded alginate sealed particles were reported to have the highest number of cells in the later stage of apoptosis or necrosis as visible in microscopic photographs Fig. 16. Discussion In this study, we have presented a comprehensive in silico and experimental investigation into potential therapeutic agents for prostate cancer. The study utilized in silico methods to identify potential drug targets associated with prostate cancer. By mining databases like DisGeNET and GeneCards, we identified a substantial number of genes linked to prostate cancer. This approach aligns with contemporary trends in bioinformatics-driven target identification, which has become increasingly crucial in precision medicine initiatives targeting cancer. A review summarizes the anticancer efficacy of quercetin against prostate cancer using PC3 and LNCaP cell lines. This study also highlighted that combination therapy of quercetin for prostate cancer treatment has various advantages such as enhanced anticancer effect, reduction in the treatment dose along with lower side effects (Yang et al., 2015 ). The binding affinity of β-Glucan and quercetin with selected targets relevant to prostate cancer, namely CYP19ABG, ABCB1, and CDK1 was determined with the help of molecular docking. The observation of a higher binding affinity of quercetin compared to β-Glucan underscores its potential as a therapeutic agent. This finding corroborates recent studies highlighting the anti-cancer properties of quercetin through various mechanisms, including apoptosis induction and inhibition of cancer cell proliferation. A study demonstrated the apoptotic effect of quercetin against a panel of 9 cancer cell lines including prostate cancer and quercetin significantly induced apoptosis in these cancer cells (Hashemzaei et al., 2017 ). We made formulation by loading quercetin into β-Glucan derived from Agaricus bisporus and yeast and alginate sealing was done for the slow and sustained release of quercetin from the hollow β-Glucan particles. The alginate-sealed formulation was reported to slow down the release of the compound in many studies (Soto et al., 2010 ). We conducted MTT assays on DU145 cells to evaluate the cytotoxic effects of particles derived from Agaricus bisporus and yeast, demonstrating a dose-dependent decrease in cell viability. Importantly, yeast-derived particles, particularly quercetin-loaded and alginate-sealed particles, exhibited promising cytotoxicity against prostate cancer cells. We found that the alginate-sealed particles i.e. ABG + Q + ALG and YBG + Q + ALG have having lowest IC50 values so they can be considered good anticancer agents against prostate cancer. We also conducted MTT on the healthy cell line RAW264.7 to assess the effect of these particles and found that particles are not affecting the viability of the cells at initial concentration. At higher concentrations such as 500µg/ml, cell viability was decreasing up to 20%. A study conducted the MTT of β-Glucan on RAW264.7 cell line and found that the control group and β-Glucan treated group did not have any significant difference in cell survival and this study aligns with our study in which we found no significant cell death after the treatment with β-Glucan and its formulations (Choi et al., 2016 ). Through fluorescence microscopy and staining techniques, we tried to find out mechanistic insights into the mode of action of the particles. The observed increase in ROS generation, alterations in lysosomal pH, decline in MMP, and induction of apoptosis highlight the significant anti-cancer potential of these compounds. Quercetin is investigated in many studies and is found to attenuate cell survival, inhibit cell proliferation pathways, and induce cell death due to the generation of ROS (Ward et al., 2018 ) and slow and sustained release of the Quercetin from the hollow β-Glucan particles due to alginate sealing will enhance its potential towards efficient anticancer activity. In this study, ROS generation increased in a dose-dependent manner that confirmed the apoptosis of the cancer cells due to oxidative stress. Nuclear fragmentation and condensation in DU145 after treatment with β-Glucan particles aligned with a study in which Farnesol was found to induce apoptosis in DU145 cells as assessed through a fluorescent microscope using DAPI staining (Park et al., 2014 ). AO/EtBr dual staining was found to show cells in the early and late apoptotic stages. AO stains live cells while EtBr stains the cells undergoing apoptosis or having compromised membranes. In a study, PAX2 siRNA knocked down DU145 cells and showed condensed yellow nuclei due to the co-localization of AO and EtBr dye (Bose et al., 2009 ). Cell scratch assay confirmed that cells are losing their capability to migrate as was studied in a study in which cyclosporine A was able to inhibit the 25–33% motility of tumor cells in DU145 cells (Cevik et al., 2019 ). This can be very beneficial because the migration of cells from one place to another can result in the metastasis of prostate cancer. AnnexinV/FITC and PI dual staining showed the cells in the early apoptotic and apoptotic/necrotic stages. AnnexinV/FITC and PI dual staining determines apoptotic cells due to the alteration in the plasma membrane. The translocation of the phosphatidyl-serine from the inner to the outer membrane leads to its binding with Annexin V showing cells in early stages of apoptosis while later stages of apoptosis allow PI to permeate into the cells due to pore formation in the plasma membrane resulting in binding with Annexin V and PI both. Quercetin-loaded and alginate-sealed particles were reported to have a significant number of cells in the later stage of apoptosis showing the efficacy of the formulation (Yusof et al., 2019). Overall, our findings suggested that particles derived from Agaricus bisporus and yeast have efficient anticancer activity and further in vivo studies and clinical investigations can give impactful findings to use these formulations for the treatment of prostate cancer. Conclusions The integrated in silico and in vitro study underscores the potential of quercetin-loaded and alginate-sealed β-Glucan particles as promising therapeutic candidates for prostate cancer treatment. Through a combination of computational target identification and in vitro experimental validation, the study elucidated the higher binding affinity of quercetin and cytotoxic effects of quercetin-loaded particles, particularly from yeast sources, against DU145 cells. ROS generation and other fluorescent assays support the anti-cancer properties of these particles. Future directions could involve preclinical studies to assess efficacy in animal models, pharmacokinetic profiling to determine bioavailability, and clinical trials to evaluate safety and efficacy in human patients. These efforts hold promise for advancing precision oncology and improving patient outcomes in prostate cancer treatment. Abbreviations ABG Agaricus bisporus derivedβ-Glucan ABG + Q Agaricus bisporus derived quercetin loadedβ-Glucan ABG + Q + ALG Agaricus bisporus derived quercetin-loaded alginate-sealedβ-Glucan YBG Yeast derivedβ-Glucan YBG + Q Yeast derived quercetin loadedβ-Glucan YBG + Q + ALG Yeast derived quercetin-loaded alginate-sealedβ-Glucan ROS Reactive oxygen species AO/EtBr Acridine orange/ethidium bromide PI Propidium Iodide Declarations Ethics approval and consent to participate Not Applicable. Consent for publication Not Applicable in this section. Availability of data and materials Will be available on request. Competing interests The authors declare no known competing interests. Funding Not Applicable Author’s contributions RT has written original draft and performed experiments. TKU conceptualized the ideas, provided resources and supervised the research. PKP provided support during research and language editing. All authors read and approved the final version of manuscript. Acknowledgements Authors are thankful to the Dr. Geetika Madan Patel, Vice President and Chairperson, Research and Development Cell, Parul University for providing laboratory facilities to conduct the proposed research work. References Atasoy Ö, Erbaş O (2020) Up to date of prostate cancer. D J Med Sci 6(2):092-102. Klusa D, Lohaus F, Furesi G, Rauner M, Benešová M, Krause M, Kurth I, Peitzsch C (2021) Metastatic spread in prostate cancer patients influencing radiotherapy response. Front. Oncol 10:627379. Hou Z, Huang S, Li Z (2021) Androgens in prostate cancer: A tale that never ends. Cancer Lett. 516:1-2. Oduwole OO, Huhtaniemi IT, Misrahi M (2021) The roles of luteinizing hormone, follicle-stimulating hormone and testosterone in spermatogenesis and folliculogenesis revisited. Int. J. Mol. Sci. 22(23):12735. Morote J, Triquell M, Campistol M, Abascal-Junquera JM, Servian P, Trilla E (2022) Stratifying the initial prostate cancer suspicion to avoid magnetic resonance exams by sequencing men according to serum prostate-specific antigen, digital rectal examination and the prostate-specific antigen density based on digital rectal prostate volume category. BJUI Compass. 4 (3): 266-8. 2022. Board PA (2023) Prostate Cancer Treatment (PDQ®). InPDQ Cancer Information Summaries [Internet]. National Cancer Institute (US). Kaur R, Sharma M, Ji D, Xu M, Agyei D (2019) Structural features, modification, and functionalities of beta-glucan. Fibers. 8(1):1. Nakajima M, Tanaka N, Motouchi S, Kobayashi K, Shimizu H, Abe K, Hosoyamada N, Abara N, Morimoto N, Hiramoto N, Nakata R (2024) Beta-Glucanase superfamily identified by sequential, functional, and structural analyses. bioRxiv. 2024-02. Kupetz M, Procopio S, Sacher B, Becker T (2015) Critical review of the methods of β-glucan analysis and its significance in the beer filtration process. Eur Food Res Technol. 1:725-36. Caseiro C, Dias JN, de Andrade Fontes CM, Bule P ((2022)) From cancer therapy to winemaking: The molecular structure and applications of β-glucans and β-1, 3-glucanases. Int. J. Mol. Sci. 23(6):3156. Sletmoen M, Stokke BT (2008) Higher order structure of (1, 3)‐β‐D‐glucans and its influence on their biological activities and complexation abilities Biopolymers: Original Research on Biomolecules. 9(4):310-21. Yang D, Wang T, Long M, Li P (2020) Quercetin: its main pharmacological activity and potential application in clinical medicine. Oxid Med Cell Longev. 2020. Ghafouri-Fard S, Shabestari FA, Vaezi S, Abak A, Shoorei H, Karimi A, Taheri M, Basiri A (2021) Emerging impact of quercetin in the treatment of prostate cancer. Biomed. Pharmacother. 138:111548. Nazemi Z, Sahraro M, Janmohammadi M, Nourbakhsh MS, Savoji H (2023) A review on tragacanth gum: A promising natural polysaccharide in drug delivery and cell therapy. Int. J. Biol. Macromol. 124343. Nam JS, Sharma AR, Nguyen LT, Chakraborty C, Sharma G, Lee SS (2016) Application of bioactive quercetin in oncotherapy: from nutrition to nanomedicine. Molecules. 21(1):108. Cao H, Wang D, Gao R, Li C, Feng Y, Chen L (2022) Therapeutic targets and signaling pathways of active components of QiLing decoction against castration-resistant prostate cancer based on network pharmacology. PeerJ. 10:e13481. Suresh NT, Ravindran VE, Krishnakumar U (2021) A computational framework to identify cross association between complex disorders by protein-protein interaction network analysis. Curr. Bioinform. 16(3):433-45. Yang L, Hu Z, Zhu J, Liang Q, Zhou H, Li J, Fan X, Zhao Z, Pan H, Fei B (2020) Systematic elucidation of the mechanism of quercetin against gastric cancer via network pharmacology approach. Biomed Res. Int. 2020. Upadhyay TK, Trivedi R, Khan F, Al-Keridis LA, Pandey P, Sharangi AB, Alshammari N, Abdullah NM, Yadav DK, Saeed M (2022) In vitro elucidation of antioxidant, antiproliferative, and apoptotic potential of yeast-derived β-1, 3-glucan particles against cervical cancer cells. Front. oncol. 12:942075. Bhosale A, Trivedi R, Upadhyay TK, Gurjar D, Aghaz F, Khan F, Pandey P, Zeyaullah M, Alam MS, Al-Najjar MA, Siddiqui S (2022) Investigation on Antimicrobial, Antioxidant, and Anti-cancerous activity of Agaricus bisporus derived β-Glucan particles against cervical cancer cell line. Cell. Mol. Biol. 68(9):150-9. Upadhyay TK, Fatima N, Sharma D, Saravanakumar V, Sharma R (2017) Preparation and characterization of beta-glucan particles containing a payload of nanoembedded rifabutin for enhanced targeted delivery to macrophages. EXCLI journal.16:210. Vodnik VV, Mojić M, Stamenović U, Otoničar M, Ajdžanović V, Maksimović-Ivanić D, Mijatović S, Marković MM, Barudžija T, Filipović B, Milošević V (2021) Development of genistein-loaded gold nanoparticles and their antitumor potential against prostate cancer cell lines. Mater. sci. eng. 124:112078. Khan F, Pandey P, Jha NK, Jafri A, Khan I (2020) Antiproliferative effect of Moringa oleifera methanolic leaf extract by down-regulation of Notch signaling in DU145 prostate cancer cells. Gene Rep. 19:100619. Zhu X, Chen X, Wang G, Lei D, Chen X, Lin K, Li M, Lin H, Li D, Zheng Q (2022) Picropodophyllin inhibits the proliferation of human prostate cancer du145 and lncap cells via ros production and pi3k/akt pathway inhibition. Biol. Pharm. Bull. 45(8):1027-35. Sana SS, Kumbhakar DV, Pasha A, Pawar SC, Grace AN, Singh RP, Nguyen VH, Le QV, Peng W (2020) Crotalaria verrucosa leaf extract mediated synthesis of zinc oxide nanoparticles: assessment of antimicrobial and anticancer activity. Molecules. 25(21):4896. Yang Q, Fang Y, Zhang C, Liu X, Wu Y, Zhang Y, Yang J, Yong K (2022) Exposure to zinc induces lysosomal-mitochondrial axis-mediated apoptosis in PK-15 cells. Ecotoxicol. Environ. Saf. 241:113716. Ning L, Zhao W, Gao H, Wu Y (2020) Hesperidin induces anticancer effects on human prostate cancer cells via ROS-mediated necrosis like cell death. J. buon. 25(6):2629-34. Fernandes NV, Yeganehjoo H, Katuru R, DeBose-Boyd RA, Morris LL, Michon R, Yu ZL, Mo H (2013) Geranylgeraniol suppresses the viability of human DU145 prostate carcinoma cells and the level of HMG CoA reductase. Exp Biol Med. 238(11):1265-74. Bobadilla AV, Arévalo J, Sarró E, Byrne HM, Maini PK, Carraro T, Balocco S, Meseguer A, Alarcón T (2019) In vitro cell migration quantification method for scratch assays. J. R. Soc. Interface. 16(151):20180709. Md Yusof EN, M. Latif MA, M. Tahir MI, Sakoff JA, Simone MI, Page AJ, Veerakumarasivam A, Tiekink ER, BSA Ravoof T (2019) o-Vanillin derived schiff bases and their organotin (IV) compounds: synthesis, structural characterisation, in-silico studies and cytotoxicity. Int. J. Mol. Sci. 20(4):854. Lin T, Liang C, Peng W, Qiu Y, Peng L (2020) Mechanisms of core Chinese herbs against colorectal cancer: a study based on data mining and network pharmacology. J. evid.-based complement. altern. 2020. Ameer K, Murtaza MA, Zainab S, Kim YM, Arshad MU, Pasha I, Abid M, Park MK. Polysaccharide-Based Films: Carriers of Active Substances and Controlled Release Characteristics. InPolysaccharide Based Films for Food Packaging: Fundamentals, Properties and Applications 2024 May 23 (pp. 379-400). Singapore: Springer Nature Singapore. Sahoo BM, Banik BK, Borah P, Jain A (2022) Reactive oxygen species (ROS): key components in cancer therapies. Anti-Cancer Agents Med. Chem (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 22(2):215-22. Kumar R, Saneja A, Panda AK (2021) An annexin V-FITC—propidium iodide-based method for detecting apoptosis in a non-small cell lung cancer cell line. Lung Cancer: Methods and Protocols. 213-23. Yang F, Song L, Wang H, Wang J, Xu Z, Xing N (2015) Quercetin in prostate cancer: Chemotherapeutic and chemopreventive effects, mechanisms and clinical application potential. Oncol. Rep. 33(6):2659-68. Hashemzaei M, Delarami Far A, Yari A, Heravi RE, Tabrizian K, Taghdisi SM, Sadegh SE, Tsarouhas K, Kouretas D, Tzanakakis G, Nikitovic D (2017) Anticancer and apoptosis‑inducing effects of quercetin in vitro and in vivo. Oncology reports. 38(2):819-28. Soto E, Kim YS, Lee J, Kornfeld H, Ostroff G (2010) Glucan particle encapsulated rifampicin for targeted delivery to macrophages. Polymers. 2(4):681-9. Choi EY, Lee SS, Hyeon JY, Choe SH, Keum BR, Lim JM, Park DC, Choi IS, Cho KK (2016) Effects of β-glucan on the release of nitric oxide by macrophages stimulated with lipopolysaccharide. Asian-Australas J Anim Sci. 29(11):1664. Ward AB, Mir H, Kapur N, Gales DN, Carriere PP, Singh S (2018) Quercetin inhibits prostate cancer by attenuating cell survival and inhibiting anti-apoptotic pathways. World J. Surg. Oncol. 16:1-2. Park JS, Kwon JK, Kim HR, Kim HJ, Kim BS, Jung JY (2014) Farnesol induces apoptosis of DU145 prostate cancer cells through the PI3K/Akt and MAPK pathways. Int. J. Mol. Med. 33(5):1169-76. Bose SK, Gibson W, Bullard RS, Donald CD (2009) PAX2 oncogene negatively regulates the expression of the host defense peptide human beta defensin-1 in prostate cancer. Mol. Immunol. 46(6):1140-8. Cevik O, Turut FA, Acidereli H, Yildirim S (2019) Cyclosporine-A induces apoptosis in human prostate cancer cells PC3 and DU145 via downregulation of COX-2 and upregulation of TGFβ. Turk Biyokim Derg. 44(1):47-54. Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 10 Jun, 2024 Reviewers invited by journal 08 Jun, 2024 Editor assigned by journal 06 Jun, 2024 First submitted to journal 05 Jun, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4486275\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":312164882,\"identity\":\"4ca90df1-399f-401b-8e1b-e8c08cc0f919\",\"order_by\":0,\"name\":\"RASHMI TRIVEDI\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABA0lEQVRIiWNgGAWjYHACNgYGAzDJIJFQYSPHDxJLKCCoxQCq5UyasWQDSIsBIS0MEBUSjG2HEzccYIALYAXm7cefPbpR8CefTyL54I0HQFuMz69O/PDAgEGeX+wAVi0yZ3LMjXMMDCzbJNKSLUB+MbvxdrME0GGGM2cnYNUiwZDDJg3UYsDGc8YM7BezG2c3gLQkGNzGoYX/+TOolvPfJBKBftk84+zmH3i1SCSYQbSw97CBtWzg792G3xaJNyAtxkAtbcYWIIdJ3ODdZpFgIIHbL/zpQIf9kTOQb2Z+ePMHKCr7z24GMeT5pbFrwWZKAiRcSAD8B0hRPQpGwSgYBSMAAAAO0lsA+3hbrQAAAABJRU5ErkJggg==\",\"orcid\":\"https://orcid.org/0000-0001-8913-5187\",\"institution\":\"Parul University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"RASHMI\",\"middleName\":\"\",\"lastName\":\"TRIVEDI\",\"suffix\":\"\"},{\"id\":312164883,\"identity\":\"a4c35167-6a41-4948-ac47-f1e56244cb6c\",\"order_by\":1,\"name\":\"Tarun Kumar Upadhyay\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0002-2551-9779\",\"institution\":\"Parul University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Tarun\",\"middleName\":\"Kumar\",\"lastName\":\"Upadhyay\",\"suffix\":\"\"},{\"id\":312164884,\"identity\":\"9cb404cd-2585-4dae-99fe-c888a06a4fb9\",\"order_by\":2,\"name\":\"Pranav Kumar Prabhakar\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Lovely Professional University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Pranav\",\"middleName\":\"Kumar\",\"lastName\":\"Prabhakar\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-05-27 16:26:22\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4486275/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4486275/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":58906477,\"identity\":\"33cf3c7e-5a5b-4aa8-8f92-0ca8711bfa0f\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:13\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":70464,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eCommon gene prediction using Venny 2.1. We have found 3 genes that were common for target and disease.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/6c1abb8230427b54b2f3d6b9.png\"},{\"id\":58906487,\"identity\":\"c2af307d-b2a4-4819-9306-1669d6497cad\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:14\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":308640,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(a) PPI network of common genes of β-Glucan, Quercetin and prostate cancer (b). Intersection of common targets of β-Glucan and quercetin with common targets of prostate cancer.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/5d31649bbafcecd182ccedab.png\"},{\"id\":58906481,\"identity\":\"ccbdc4ef-3c43-4aa5-8d96-5cc06bc75e49\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:13\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":348308,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(a and b) are the interaction of ABCB1 with Quercetin and β-Glucan respectively. (b and c) are interaction of CDK1 with Quercetin and β-Glucan respectively while (e and f) are interaction of CYP19ABG with Quercetin and β-Glucan respectively.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/7005553f402cd455656ce055.png\"},{\"id\":58907237,\"identity\":\"5fc39403-b597-41a2-aeb2-9a7a3eb2c175\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:16:13\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":184894,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eHollow β-Glucan particle having pores were loaded with quercetin and further sealed with sodium alginate.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/0542b905c1387ba70007b120.png\"},{\"id\":58906483,\"identity\":\"12c84d68-1bdc-426e-aba3-0c624e55209f\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:14\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":53282,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMTT of the particles derived from \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e β-Glucan. We found a decrease in cell viability in dose dependent manner after treatment (a) % Viability of DU145 cells after treatment with ABG (b) % Viability of DU145 cells after treatment with Quercetin (c) % Viability of DU145 cells after treatment with ABG+Q (d) % Viability of DU145 cells after treatment with ABG+Q+Alg.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/c3e7dcf395b0b27ce21e4643.png\"},{\"id\":58906479,\"identity\":\"ef7ea2d0-6c28-4a39-af14-69fb367a0ba5\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:13\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":61388,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMTT of the particles derived from yeast β-Glucan. There was dose dependent reduction in cell viability after treatment (a) % Viability of DU145 cells after treatment with YBG (b) Cell viability of DU145 after treatment with Quercetin (c) % Viability of DU145 cells after treatment with YBG+Q (d) % Viability of DU145 cells after treatment with YBG+Q+Alg.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/70709b44a52fecf4af4da205.png\"},{\"id\":58906482,\"identity\":\"89f42f6d-a516-4334-aaf8-8d02e275f20b\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:13\",\"extension\":\"png\",\"order_by\":7,\"title\":\"Figure 7\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":79595,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEvaluation of the cell viability with MTT on RAW264.7 cells. (a, b, c) are the cell viability of the cells treated with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles, (d, e, f) are the cell viability of the cells treated with yeast derived particle while (g) is the cell viability of the cells treated with quercetin.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"7.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/5422b4e557992a03a93a829f.png\"},{\"id\":58906478,\"identity\":\"6b7d349f-ab15-4883-8607-5e49fccf8c2f\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:13\",\"extension\":\"png\",\"order_by\":8,\"title\":\"Figure 8\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":422124,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDU145 cells were showing visible morphological changes after treatment for 24 h (i) DU145 cells after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. (ii) DU145 cells after treatment with yeast derived particles.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"8.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/ecd368557e6a688e463c0ee8.png\"},{\"id\":58906488,\"identity\":\"b8d4e74f-a2bf-4531-8b7a-92537a11ff37\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:15\",\"extension\":\"png\",\"order_by\":9,\"title\":\"Figure 9\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":365210,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(i) DU145 cells showing efficient amount of ROS generation after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. (ii) DU145 cells after treatment with yeast derived particles. (a, b, c, d) are the statistical analysis of the ABG, Quercetin, ABG+Q, and ABG+Q+Alg. (e, f, g, h) are the statistical analysis of the YBG, Quercetin, YBG+Q+Alg, and YBG+Q+Alg. All the values are showing high significance with p value less than 0.0005 (\\u003cem\\u003ep\\u0026lt;0.0005\\u003c/em\\u003e)\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"9.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/68b0e9462f3aa291c70e9d8d.png\"},{\"id\":58906490,\"identity\":\"3fca2988-b60b-400b-9c1f-cb84c610c1c7\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:15\",\"extension\":\"png\",\"order_by\":10,\"title\":\"Figure 10\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":372624,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMorphological changes in nucleus observed with the DAPI staining for the (i) \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e and (ii) Yeast derived β-Glucan particles.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"10.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/4ed0d9de2cb5fe6cade550f1.png\"},{\"id\":58906491,\"identity\":\"34848f98-6b6e-4a57-a83e-35e9e3ec0a58\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:15\",\"extension\":\"png\",\"order_by\":11,\"title\":\"Figure 11\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":311010,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eApoptosis determination in the DU145 cells as observed with the FLoid imaging microscope. (i) DU145 cells after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. (ii) DU145 cells after treatment with yeast derived particles. (a, b, c, d) are the statistical analysis of the ABG, Quercetin, ABG+Q, and ABG+Q+ALG. (e, f, g, h) are the statistical analysis of the YBG, Quercetin, YBG+Q, and YBG+Q+ALG. All the values were highly significant except (g) which was less significant with p value less than 0.05 (\\u003cem\\u003ep\\u0026lt;0.05\\u003c/em\\u003e)\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"11.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/943808988e41b225a6607fa3.png\"},{\"id\":58906492,\"identity\":\"58b9b59f-35ac-463a-bc27-27d2c51de6af\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:16\",\"extension\":\"png\",\"order_by\":12,\"title\":\"Figure 12\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":472102,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLysoTracker red DND-99 staining in the DU145 cells observed with the fluorescence microscope (i) DU145 cells after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. (ii) DU145 cells after treatment with yeast derived particles. (a, b, c, d) are the statistical analysis of the ABG, Quercetin, ABG+Q, and ABG+Q+ALG. (e, f, g, h) are the statistical analysis of the YBG, Quercetin, YBG+Q, and YBG+Q+ALG. (a), (c) and (d) were highly significant with p value less than 0.05 (\\u003cem\\u003ep\\u0026lt;0.05\\u003c/em\\u003e) while (b) was found to be less significant with (\\u003cem\\u003ep\\u0026gt;0.05\\u003c/em\\u003e). (e) and (f) were highly significant while (g) and (h) were less significant.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"12.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/4d1bcb6f3fddce82a61f0508.png\"},{\"id\":58906485,\"identity\":\"f7fadcde-1fec-43d5-b794-b0788e3dbbe3\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:14\",\"extension\":\"png\",\"order_by\":13,\"title\":\"Figure 13\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":421641,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMitoTracker Red staining in the DU145 cells as observed with the fluorescence microscope for the determination of mitochondrial activity. (i) DU145 cells after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. (ii) DU145 cells after treatment with yeast derived particles. (a, b, c, d) are the statistical analysis of the ABG, Quercetin, ABG+Q, and ABG+Q+ALG. (e, f, g, h) are the statistical analysis of the YBG, Quercetin, YBG+Q, and YBG+Q+ALG. (a) (b) and (c) are highly significant with p value less than 0.05 (\\u003cem\\u003ep\\u0026lt;0.05\\u003c/em\\u003e) while (d) was less significant with (\\u003cem\\u003ep\\u0026lt;0.05\\u003c/em\\u003e). (e), (g) and (h) were highly significant while (f) was less significant.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"13.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/f4e331adcfd334d75c5c5b1f.png\"},{\"id\":58906486,\"identity\":\"3c543cbe-2337-45a5-aa56-6fdc8ea0c1e8\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:14\",\"extension\":\"png\",\"order_by\":14,\"title\":\"Figure 14\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":487078,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eAO/EtBr dual staining of the DU145 cells showing cells in light yellowish color cells are in early and orange color of cells in the late stage of the apoptosis. Cells showing yellow color of fluorescence are in their initial stage of apoptosis while cells in yellow-orange-red fluorescence are in later stages of apoptosis.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"14.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/21a53defb40416f3f8084a41.png\"},{\"id\":58906484,\"identity\":\"8736095a-012e-40ff-ba22-b4e19d5448f9\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:14\",\"extension\":\"png\",\"order_by\":15,\"title\":\"Figure 15\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1295589,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eThe progress of wound closure in control wells indicating the natural rate of cell migration and proliferation under normal conditions. In the treated wells, there was a significant 10 to 15%.increase in the area of the wound compared to the control wells.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"15.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/43ad04719bdf8297f62e9e3e.png\"},{\"id\":58906489,\"identity\":\"490de48d-7a0e-461d-a849-a477f36a91f9\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:08:15\",\"extension\":\"png\",\"order_by\":16,\"title\":\"Figure 16\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":199951,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eAnnexin V/ FITC and PI dual staining showing the maximum apoptosis in the cells treated with the quercetin-loaded alginate-sealed particles derived from \\u003cem\\u003eAgaricus bisporus \\u003c/em\\u003eand yeast.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"16.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/0afe3aa4c6753f4986918614.png\"},{\"id\":58907584,\"identity\":\"a34b4ba6-2b43-4024-9ab9-11bc6cc94a46\",\"added_by\":\"auto\",\"created_at\":\"2024-06-24 03:24:17\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":6292830,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4486275/v1/dd3d81c0-652d-4eaa-a2dd-b3bba64c16df.pdf\"}],\"financialInterests\":\"\",\"formattedTitle\":\"Quercetin loaded and alginate sealed β-Glucan particles-based drug delivery system against DU145 a prostate cancer cell line: Integrating network pharmacology, molecular docking and in vitro studies\",\"fulltext\":[{\"header\":\"Background\",\"content\":\"\\u003cp\\u003eProstate cancer is a tumor that occurs in the prostate gland due to uncontrolled growth of the cells and represents a huge challenge in the landscape of oncology. When cells of prostate cancer start to metastasize to different organs, the survival rate of the patient starts to decline significantly (Atasoy and Erbaş \\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). The process of metastasis and the tendency of prostate cancer cells to target specific organs are influenced by various cellular subtypes and the unique microenvironments of those organs, along with the interactions between them (Klusa et al., \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). The androgen hormone, testosterone, and its metabolite dihydrotestosterone (DHT) play pivotal roles in prostate cancer development and progression. Binding of androgen to the androgen receptor leads to the activation of cell proliferation signaling leading to the development of prostate cancer. Androgen receptor signaling pathways lead to the proliferation and survival of prostate cells in the malignant stage, making androgen deprivation therapy (ADT) through the removal of gonads a cornerstone in the management of advanced disease (Hou et al., \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). Gonadotropin-releasing hormone (GnRH) is found to be involved in the inhibition of steroidogenesis leading to the suppression of androgen receptors resulting in good management of prostate cancer (Oduwole et al., \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). Symptoms of prostate cancer are nonspecific or may remain asymptomatic until later stages. Screening strategies, including serum prostate-specific antigen (PSA) and digital rectal examination (DRE) measurement, are the techniques that are used for the early-stage detection of prostate cancer (Morote et al., \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Currently available treatment protocols for prostate cancer, including surgical interventions, radiation therapy, hormone therapy, chemotherapy, and immunotherapy, offer valuable options for patients still, each treatment comes with its set of limitations and potential side effects that can significantly impact well-being and quality of life of the patient that leads to the continued research in prostate cancer treatment (Board \\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Surgical interventions, such as radical prostatectomy, are effective in removing localized tumors, however, they can lead to complications such as urinary incontinence, erectile dysfunction, and bowel dysfunction.\\u003c/p\\u003e \\u003cp\\u003ePolysaccharides are a digestible and indispensable component present in a variety of foods that provide essential calories and glucose necessary for regular bodily functions along with various health benefits. One extensively studied and well-documented bioactive polysaccharide is β-Glucan having monomer units linked through glycosidic bonds at β 1\\u0026rarr;3, 1\\u0026rarr;4, and/or 1\\u0026rarr;6 positions, forming branched or unbranched structures (Kaur et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). β-Glucans are carbohydrate polymers and they are the main constituents of the cell walls of algae, lichens, yeasts, fungi, and bacteria, as well as in many plants and have a molecular weight in the range between 50 kDa to 4100 kDa contributing to their diverse biological activities (Nakajima et al., \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e2024\\u003c/span\\u003e). β-Glucans exhibit a rigid structure characterized by β-1\\u0026rarr;3 and 1\\u0026rarr;6) glycosidic linkage in the case of yeast and mushrooms, whereas β-1\\u0026rarr;3 and 1\\u0026rarr;4) glycosidic linkages in cereals. The molecular weight of β-Glucans is influenced by various factors, including the type of detectors utilized, reaction conditions, solvents employed, and the specific compounds present in the sample. These parameters collectively determine the size and distribution of β-Glucan molecules, impacting their functional properties and physiological effects (Kupetz et al., \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). β-Glucans exhibit diverse biological activities influenced by several key parameters. These include molecular weight, structural characteristics, conformational arrangement such as single helix, triple helix, or random coil, charge on the polymer, and density of branching. Changes or modifications in the conformation of β-Glucans can profoundly impact their immune-modulatory properties and overall functionality (Caseiro et al., \\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Despite being insoluble in water, β-Glucans find extensive applications in the food and pharmaceutical industries. They serve as valuable raw materials for various beverages and medicinal products, contributing to the therapeutics for numerous diseases including cholesterol management, diabetes, and cancer (Sletmoen and Stokke \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eQuercetin is a flavonoid in fruits and veggies, having a ketocarbonyl group and four active hydroxyl groups with scientific name 3, 3', 4', 5, 7-pentahydroxyflavone. The phenolic hydroxyl group and double bond presence make it a strong antioxidant (Yang et al., \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). It boosts anti-inflammatory, antioxidant, and anticancer properties along with offering vasodilatory, anti-hypertensive, anti-hypercholesterolemic, anti-obesity, and anti-atherosclerotic effects. Its standout quality is triggering tumor cell death (apoptosis), hindering the progression of various human cancers like prostate cancer while sparing normal cells. Specifically with prostate cancer, quercetin indirectly hampers key genes like the prostate-specific antigen (PSA), and androgen receptor (AR) potentially impeding cancer development (Fard et al., 2021). Although quercetin is reported to have various biological activities, its lower solubility is one of its drawbacks in medicine so many researchers tried to load it in the nano/microparticles for its targeted delivery and found positive more effective, and improved biological activity (Nazemi et al., \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Quercetin has the potential for the treatment of cancer but it has some limitations such as low aqueous solubility, getting clear very rapidly from the body, high metabolic rate, and very poor absorption that make it restricted for use in cancer therapy. To overcome these limitations, the use of a delivery agent/carrier may enhance its absorption, solubility, and target specificity (Nam et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e). In a variety of cancers including prostate cancer, Quercetin is found to inhibit a variety of enzymes that are responsible for the activation of carcinogens and cell proliferation signaling pathways. In this study, we have loaded Quercetin in the hollow β-Glucan particles and sealed it with alginate for the slow and sustained release of the Quercetin for the possible treatment of prostate cancer. The novelty of the study lies in the innovative approach of encapsulating quercetin within a β-Glucan carrier and then sealing it with sodium alginate for targeted therapy against prostate cancer. This method addresses several limitations associated with the use of quercetin in cancer therapy, including its low aqueous solubility, rapid clearance, high metabolic rate, and poor absorption.\\u003c/p\\u003e\"},{\"header\":\"Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eReagents and chemicals\\u003c/h2\\u003e \\u003cp\\u003eQuercetin (HiMedia), MTT, DAPI, PI, H2DCFDA, LysoTracker Red DND 99, MitoTracker Red CMX-ROS, Acridine orange, Ethidium bromide, Annexin V/FITC were purchased from Invitrogen (Thermo Fisher Scientific). DMEM (HiMedia), fetal bovine serum (FBS), antibiotic, and antimycotic solution, were purchased from Gibco\\u0026trade; .\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMaintenance of the Cell culture\\u003c/h2\\u003e \\u003cp\\u003eDU145 cell line for the prostate cancer was obtained from the National Centre for Cell Science (NCCS) Pune, India. Cells were maintained at 37\\u0026deg;C and 5% CO2 in a CO\\u003csub\\u003e2\\u003c/sub\\u003e incubator. Complete DMEM media was supplemented with 10% FBS and 1% antibiotic and antimycotic solution.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eIn silico\\u003c/b\\u003e \\u003cb\\u003estudies\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProstate cancer related gene identification\\u003c/h2\\u003e \\u003cp\\u003eTarget genes related to prostate cancer were identified by exploring the term prostate cancer in the GeneCards (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://www.genecards.org/\\u003c/span\\u003e\\u003cspan address=\\\"https://www.genecards.org/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e) and DisGeNET (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://www.disgenet.org/\\u003c/span\\u003e\\u003cspan address=\\\"https://www.disgenet.org/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e) databases and different targets were downloaded into Excel format (Cao et al., \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). The targets from two databases pooled and duplicates were removed to obtain the final targets for the prostate cancer.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eCollection of drug targets\\u003c/h2\\u003e \\u003cp\\u003eWe have selected common targets between β-Glucan and Quercetin using Swiss target prediction software. Common targets related to β-Glucan, Quercetin, and prostate cancer were recognized using Venny 2.1 (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://bioinfogp.cnb.csic.es/tools/venny/\\u003c/span\\u003e\\u003cspan address=\\\"https://bioinfogp.cnb.csic.es/tools/venny/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePPI Network construction\\u003c/h2\\u003e \\u003cp\\u003eProtein-Protein Interaction (PPI) network was analyzed using STRING 11.0 database (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://string-db.org/\\u003c/span\\u003e\\u003cspan address=\\\"https://string-db.org/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e) (Suresh et al., \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). These common potential target genes were merged into the STRING 11.0 database to create a PPI network, species keeping to Homo sapiens.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMolecular docking\\u003c/h2\\u003e \\u003cp\\u003eLigand β-Glucan and Quercetin were imported from PubChem databases. Macromolecules were downloaded in PDB format from RCSB PDB (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://www.rcsb.org/\\u003c/span\\u003e\\u003cspan address=\\\"https://www.rcsb.org/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e). Molecular docking was performed using CB Dock2 online server (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://cadd.labshare.cn/cb-dock2/index.php\\u003c/span\\u003e\\u003cspan address=\\\"https://cadd.labshare.cn/cb-dock2/index.php\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e) (Yang et al., \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eIn vitro\\u003c/b\\u003e \\u003cb\\u003eanticancer activity determination\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eParticle preparation and characterization\\u003c/h2\\u003e \\u003cp\\u003eβ-Glucan particles were prepared using the acid-base extraction method from the Agaricus bisporus and yeast and characterization was performed using FTIR analysis. We have already published the preparation and characterization methodologies (Upadhyay et al., \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Bhosale et al., \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Loading of quercetin into β-Glucan and alginate sealing was performed according to the previously published protocol (Upadhyay et al., \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAssessment of the cell viability of DU145 and RAW264.7 cells with MTT assay\\u003c/h2\\u003e \\u003cp\\u003eWe have assessed the cell viability using the previously described protocol (Vodnik et al., \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). 10,000 cells DU145 and RAW264.7 (healthy cell line) were placed in each well of a 96-well plate and left to grow for 24 hours in a humid environment with 5% CO2 at 37\\u0026deg;C. Following this, the cells were exposed to particles prepared in concentrations of IC50 and below and above IC50 for another 24 hours. After treatment, the old media was removed, and MTT dye (10 \\u0026micro;l) was added to each well, followed by an incubation of 4 h at a temperature of 37\\u0026deg;C. Next, 100 \\u0026micro;l DMSO was added to solubilize the formazan crystals formed by viable cells, and after 10 minutes, the absorbance of the dissolved crystals was computed at 490 nm. Cell viability was then calculated using a provided formula\\u003cdiv id=\\\"Equa\\\" class=\\\"Equation\\\"\\u003e\\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equa\\\" name=\\\"EquationSource\\\"\\u003e\\n$$\\\\text{%}\\\\text{V}\\\\text{i}\\\\text{a}\\\\text{b}\\\\text{i}\\\\text{l}\\\\text{i}\\\\text{t}\\\\text{y}=\\\\left(\\\\frac{\\\\text{A}\\\\text{b}\\\\text{s}\\\\text{o}\\\\text{r}\\\\text{b}\\\\text{a}\\\\text{n}\\\\text{c}\\\\text{e} \\\\text{o}\\\\text{f} \\\\text{t}\\\\text{r}\\\\text{e}\\\\text{a}\\\\text{t}\\\\text{m}\\\\text{e}\\\\text{n}\\\\text{t}}{\\\\text{A}\\\\text{b}\\\\text{s}\\\\text{o}\\\\text{r}\\\\text{b}\\\\text{a}\\\\text{n}\\\\text{c}\\\\text{e} \\\\text{o}\\\\text{f} \\\\text{c}\\\\text{o}\\\\text{n}\\\\text{t}\\\\text{r}\\\\text{o}\\\\text{l}}\\\\right)\\\\text{X}100$$\\u003c/div\\u003e\\u003c/div\\u003e\\u003c/p\\u003e \\u003cp\\u003eConcentrations of IC\\u003csub\\u003e50\\u003c/sub\\u003e and below and above IC\\u003csub\\u003e50\\u003c/sub\\u003e were selected for the further analysis.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMorphological observations\\u003c/h2\\u003e \\u003cp\\u003eMorphological changes in the treated cells were observed as per the previously defined protocol (Khan et al., \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). In brief, 50,000 cells/well were added in a 24-well plate and left overnight for the attachment. After 24 h, cells were given treatment with the selected concentrations of the particles and Quercetin and further left for incubation for 24 h at 37oC. After incubation, cells were washed with the PBS, and morphological changes were observed and captured with the help of an EVOS FLoid imaging station.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAssessment of Reactive oxygen species (ROS) generation\\u003c/h2\\u003e \\u003cp\\u003eThe assessment of reactive oxygen species (ROS) production after particle treatment was conducted according to the protocol described by (Zhu et al., \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). Initially, a cell density of 50,000 cells per well was seeded into a 24-well plate and permitted to incubate for 24 hours to ensure attachment. Following this, the cells were treated with selected concentrations of β-Glucan and Quercetin and then maintained in culture for an additional 24 hours at 37\\u0026deg;C with 5% CO2. After the incubation period, the cells were rinsed through phosphate-buffered saline (PBS) and stained with 20\\u0026micro;M DCFDA dye, followed by incubation at 37\\u0026deg;C for 20 minutes. Finally, images were captured using an EVOS FLoid imaging station.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eDetermination of nuclear morphology and DNA fragmentation with DAPI staining\\u003c/h2\\u003e \\u003cp\\u003eDAPI staining was carried out based on a previously described protocol with slight adjustments, as outlined in the work by (Sana et al., \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). In brief, 50,000 cells/ well were seeded in a 24-well plate and allowed to incubate for 24 h. Following this, the cells were exposed to selected concentrations of the particles and further incubated for 24 h, at temperature 37\\u0026deg;C. Once the incubation period was completed, the cells were rinsed with PBS and the cells were stained with 1\\u0026micro;g/mL of DAPI for 15 minutes. Morphological changes were then observed and documented.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eApoptosis detection with Propidium iodide (PI) staining\\u003c/h2\\u003e \\u003cp\\u003eCells undergoing apoptosis were determined using PI staining with the help of an already established protocol with minor modification (Yang et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). In Brief, 50,000 cells per well were seeded in a 24-well plate and incubated overnight. After that, cells were treated with a selected concentration of Quercetin and prepared particles and further incubated for 24 h. After incubation, cells were stained with 1\\u0026micro;g/ml of the PI (prepared from 1mg/ml of PI) and further incubated for 10 min at 37oC following which images were obtained with the EVOS FLoid imaging station.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAcidic organelles activity analysis through LysoTracker Red DND-99\\u003c/h2\\u003e \\u003cp\\u003eAcidic organelles were detected by using LysoTracker Red (100nM) as per the formerly described method (Ning et al., \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). In brief, 50,000 cells/well seeded in 24 well plate and incubated overnight. After the attachment of the cells, treatment with the various concentrations of particles was performed and further incubated for 24 h. Post incubation, cells were washed with PBS and stained with LysoTracker for 30 min and imaged under a FLoid imaging microscope.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMitochondrial membrane potential (MMP) assessment\\u003c/h2\\u003e \\u003cp\\u003eMMP was assessed as per the previously reported protocol (Fernandes et al., \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e). Briefly, 50,000 cells/well were seeded in 24 well plate and incubated for 24 h, afterwards, cells were treated with the concentrations of particles and further incubated for 24 hrs. After incubation, cells were stained for 30 min using the dye MitoTracker red CMX ROS (300nM) and imaged under EVOS FLoid imaging microscope.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec17\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAcridine orange and Ethidium bromide (AO/EtBr) dual staining\\u003c/h2\\u003e \\u003cp\\u003eAO/EtBr dual staining was performed conferring to the earlier described protocol with slight modification (Bobadilla et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). 50,000 cells/ well seeded in 24 well plate and after gaining confluency, treated with selected concentrations of particles and left for 24 h incubation in a CO2 incubator. After that, cells were stained with 5 \\u0026micro;l of AO and 5 \\u0026micro;l EtBr (5mg/ml each), and cell death was observed with a FLoid Imaging microscope.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec18\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eTo check anticancer activity via Wound healing/Cells Scratch assay\\u003c/h2\\u003e \\u003cp\\u003eTo determine the scratch healing ability, DU145 cells were seeded at a density of 50000 cells/ well in a 24-well plate and left overnight for adhesion. Before treatment, a scratch was made with the help of a sterile 10\\u0026micro;l tip and cells were imaged followed by treatment with different doses of the particles. After 24 h, cells were imaged again to quantify the change in scratch area under an inverted microscope. The area of the scratch or wound was quantified by the given formulae (Yusof et al., 2019).\\u003cdiv id=\\\"Equb\\\" class=\\\"Equation\\\"\\u003e\\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equb\\\" name=\\\"EquationSource\\\"\\u003e\\n$$\\\\text{W}\\\\text{o}\\\\text{u}\\\\text{n}\\\\text{d} \\\\text{a}\\\\text{r}\\\\text{e}\\\\text{a} \\\\text{p}\\\\text{e}\\\\text{r}\\\\text{c}\\\\text{e}\\\\text{n}\\\\text{t}=\\\\frac{\\\\text{A}\\\\text{t}}{\\\\text{A}0}\\\\text{x}100$$\\u003c/div\\u003e\\u003c/div\\u003e\\u003c/p\\u003e \\u003cp\\u003eWhere A\\u003csub\\u003et\\u003c/sub\\u003e= wound area after treatment and A\\u003csub\\u003e0\\u003c/sub\\u003e\\u0026thinsp;=\\u0026thinsp;Wound area before treatment.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAnnexinV/FITC and Propidium iodide dual staining\\u003c/h2\\u003e \\u003cp\\u003eApoptotic cells were analyzed using AnnexinV/FITC and PI dual staining as per the previously reported protocol. Briefly, DU145 cells were seeded at a density of 50000 cells/well in a 24-well plate and left overnight for the incubation. After the attachment of the cells, they were treated with IC50 concentrations of the prepared particles and left for another 24 h. After incubation, cells were washed through PBS and stained with 1\\u0026micro;l of AnnexinV/FITC and PI each and images under a fluorescence microscope (Lin et al., \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec20\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eStatistical analysis\\u003c/h2\\u003e \\u003cp\\u003e \\u003cem\\u003eIn vitro\\u003c/em\\u003e experiments were accomplished in triplicates and One-way ANOVA was used for the statistical analysis using GraphPad Prism 8.0 and ImageJ. A probability value of p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05 was deliberated statistically significant where *** means highly significant, ** means less significant and * means least significant.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e \\u003cb\\u003eIn silico\\u003c/b\\u003e \\u003cb\\u003estudies\\u003c/b\\u003e\\u003c/p\\u003e \\u003cdiv id=\\\"Sec22\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProstate cancer related gene identification\\u003c/h2\\u003e \\u003cp\\u003e683 gene were retrieved from DisGeNET while 13832 genes were identified from GeneCards. Common genes were retrieved using Venny 2.1 after removing duplicate genes.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec23\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eCollection of drug targets\\u003c/h2\\u003e \\u003cp\\u003eDrug targets were retrieved using Swiss target prediction software. 100 targets were downloaded as comma separated value for both β-Glucan and quercetin. Common targets were retrieved using Venny 2.1 as shown in Fig.\\u0026nbsp;1.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec24\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eConstruction of PPI Network\\u003c/h2\\u003e \\u003cp\\u003ePPI network of common genes of β-Glucan, Quercetin and prostate cancer is shown in Fig.\\u0026nbsp;2a. After intersection of common targets of β-Glucan and quercetin with common targets of prostate cancer, we obtained 3 gene as shown in Fig.\\u0026nbsp;2b that we have taken for the molecular docking.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec25\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eMolecular docking\\u003c/h2\\u003e \\u003cp\\u003eLigand β-Glucan and Quercetin were docked using CB dock2 with CYP19ABG (PDB ID: 5JKW), ABCB1 (PDB ID: 6QEX), and CDK1 (PDB ID: 6TWN). Interaction of ligands and targets is shown in Fig.\\u0026nbsp;3. In each docking, quercetin shows higher binding affinity with the targets than β-Glucan. The binding energies of ligands and targets and their interacting amino acids are presented in Table\\u0026nbsp;1.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eBinding energies and interacting amino acids of the ligand and target docking.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS. No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTarget\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eLigand\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eBinding energy\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eInteracting Amino acids\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eABCB1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eQuercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-8.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: TRP232 ILE235 LEU236 SER238 PHE239 THR240 ASP241 LYS242 GLU243 LEU244 ALABG\\u0026thinsp;+\\u0026thinsp;Q46 TYR247 ALABG\\u0026thinsp;+\\u0026thinsp;Q84 LYS285 GLYBG\\u0026thinsp;+\\u0026thinsp;Q88 ILE289 LYS291 ALABG\\u0026thinsp;+\\u0026thinsp;Q92 ALABG\\u0026thinsp;+\\u0026thinsp;Q95 ASN296 ILE299 PHE303 PHE770 GLN773 GLY774 PHE777 GLY778 ALA780 GLY781 GLU782 THR785 LYS786 ARG789 ALA823 LYS826 GLY830 SER831 ALA834 VAL835 GLN838 MET876 GLN990 VAL991 PHE994 ALA995 PRO996\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eβ-Glucan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-7.9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: PHE239 THR240 ASP241 LYS242 GLU243 LEU244 ALABG\\u0026thinsp;+\\u0026thinsp;Q46 TYR247 ALABG\\u0026thinsp;+\\u0026thinsp;Q84 LYS285 ILE287 GLYBG\\u0026thinsp;+\\u0026thinsp;Q88 ILE289 LYS291 ALABG\\u0026thinsp;+\\u0026thinsp;Q92 ALABG\\u0026thinsp;+\\u0026thinsp;Q95 ASN296 PHE770 GLN773 GLY774 PHE775 PHE777 GLY778 LYS779 ALA780 GLY781 GLU782 ILE783 THR785 LYS786 ARG789 ALA823 LYS826 GLY827 GLY830 SER831 ALA834 VAL835 GLN838 VAL991 PHE994 ALA995 PRO996\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCDK1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eQuercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-8.0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: THR1465 ALABG468 ARG1469 GLN1472\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003eChain B\\u003c/b\\u003e: LEU1367 LEU1370 GLU1371 ARG1374 GLU1375 GLU1378 LEU1607 GLU1608 ALABG610 GLYBG611 GLYBG612\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eβ-Glucan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-7.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: THR1465 ALABG468 ARG1469 GLN1472\\u003c/p\\u003e \\u003cp\\u003e\\u003cb\\u003eChain B\\u003c/b\\u003e: LEU1367 LEU1370 GLU1371 THR1372 ARG1374 GLU1375 GLU1378 LYS1604 LEU1607 GLU1608 SER1609 GLYBG611 GLYBG612 GLN1615\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003e3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCYP19ABG\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eQuercetin\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-7.8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: ARG115 ILE132 ILE133 PHE134 ARG192 SER199 PHE203 GLN218 PHE221 ASP222 TRP224 GLN225 GLU302 MET303 ILE305 ALABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG06 ALABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG07 PRO308 ASP309 THR310 MET311 SER312 VAL313 SER314 MET364 VAL369 VAL370 LEU372 VAL373 MET374 ARG375 PRO429 PHE430 GLY436 CYS437 ALA438 GLY439 ILE442 ALA443 MET446 MET447 ILE474 LEU477 SER478 LEU479 HIS480 PRO481 ASP482 GLU483 THR484\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eβ-Glucan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e-7.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eChain A\\u003c/b\\u003e: ARG115 ILE132 ILE133 PHE134 TRP141 ARG145 PHE148 LEU152 LEU188 ARG192 PHE203 ILE217 GLN218 GLYBG\\u0026thinsp;+\\u0026thinsp;Q19 PHE221 ASP222 TRP224 GLN225 ALABG\\u0026thinsp;+\\u0026thinsp;Q26 GLU302 MET303 ALABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG06 ALABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG07 PRO308 ASP309 THR310 MET311 SER312 VAL313 SER314 MET364 VAL369 VAL370 LEU372 VAL373 MET374 ARG375 ILE398 PRO429 PHE430 GLY431 ARG435 GLY436 CYS437 ALA438 GLY439 LYS440 ILE442 ALA443 MET446 MET447 ILE474 LEU477 SER478 HIS480 PRO481 ASP482 GLU483 THR484\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec28\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eDetermination of anticancer activity\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec29\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eParticle preparation\\u003c/h2\\u003e \\u003cp\\u003ePrepared β-Glucan particles from the \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e and yeast, loaded with quercetin and alginate sealed were used for the determination of \\u003cem\\u003ein vitro\\u003c/em\\u003e anti-cancer activity as shown in Fig.\\u0026nbsp;4. We hypothesized the slow and sustained release of the quercetin from β-Glucan after alginate sealing as is reported in many studies (Ameer et al., 2024). Hollow β-Glucan from \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e and yeast were named ABG and YBG, Quercetin loaded were named ABG\\u0026thinsp;+\\u0026thinsp;Q, YBG\\u0026thinsp;+\\u0026thinsp;Q, and Alginate sealed particles were named ABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG, YBG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG respectively. Q is used for quercetin here.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eAssessment of the cell viability with MTT assay on DU145 and RAW264.7 cells\\u003c/h3\\u003e\\n\\u003cp\\u003eCell viability of the DU145 cells upon exposure to the particles derived from Agaricus bisporus and yeast was found to decrease as shown in Fig.\\u0026nbsp;5 and Fig.\\u0026nbsp;6. Yeast-derived particles were found to have lower IC50 than the \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived particles. In the case of yeast-derived particles, quercetin-loaded-alginate-sealed particles have the lowest IC50. Concentrations of IC50 and below and above IC50 were taken for the further microscopic examination of anticancer activity. To evaluate the consequence of the particles on the healthy cells we treated RAW264.7 cells with the serial dilution from 500\\u0026micro;g/ml to 1.56\\u0026micro;g/ml. We found that particles did not have cytotoxic effects at initial concentrations as shown in Fig.\\u0026nbsp;7 and at higher concentrations of 500 and 250\\u0026micro;g/ml, particles had very little cytotoxic effects on the RAW264.7 cells.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec34\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eMorphological observations\\u003c/h2\\u003e \\u003cp\\u003eWith the help of FLoid imaging microscope, we observed clear morphological changes in the shape of the DU145 cells. At the highest concentration, cells were highly disrupted and they lost their actual shape as shown in Fig.\\u0026nbsp;8.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eAssessment of Reactive oxygen species (ROS) generation\\u003c/h3\\u003e\\n\\u003cp\\u003eROS are a natural derivative of oxygen metabolism in the body and have a variety of important roles in cell signaling. However, excessive ROS generation can lead to oxidative damage leading to the apoptosis of the cancer cells (Sahoo et al., \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e). In this study, we found that β-Glucan and the quercetin loaded β-Glucan particles have the potential to generate an efficient amount of ROS in DU145 cells as shown in Fig.\\u0026nbsp;9.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e\\n\\u003ch3\\u003eDetermination of nuclear morphology and DNA fragmentation with DAPI staining\\u003c/h3\\u003e\\n\\u003cp\\u003eThe influence of particles on the DU145 cells was assessed with DAPI staining that is specific for the binding with the nucleus. Nuclear condensation and abnormalities are quite observable and can be seen in Fig.\\u0026nbsp;10. We have found that nuclear deformities were increasing with the increase in the treatment concentration of the particles.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec37\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eApoptosis detection with Propidium iodide (PI) staining\\u003c/h2\\u003e \\u003cp\\u003ePI is a very specific dye that only binds with the dying or apoptotic cells resulting in red color fluorescence. Cells in the late apoptotic or early necrotic stage stain red. In our study we have found a significant increase in the apoptotic cells with the increase in concentration as shown in Fig.\\u0026nbsp;11. In non-treated (control) cells, there was no significant percentage of apoptosis was observed.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec38\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eLysoTracker Red DND-99 staining to monitor pH change in lysosome\\u003c/h2\\u003e \\u003cp\\u003eLysosome functioning depends on the lysosomal pH and a decrease in the fluorescence intensity of the red color indicates the elevation in the pH of lysosomes. We have found a dose-dependent decrease in the fluorescence intensity indicating the pH alteration that can result in the apoptosis of the cells as shown in Fig.\\u0026nbsp;12.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec39\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMitochondrial membrane potential determination\\u003c/h2\\u003e \\u003cp\\u003eMitoTracker Red CMX-ROS is a probe-based cationic dye that enables to assess the change in MMP. In our study, DU145 cells showed significant decline in MMP after treatment with \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e and yeast derived particles as shown in Fig.\\u0026nbsp;13.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eAcridine orange and Ethidium bromide (AO/EtBr) staining for the detection of cells in early and late stage of apoptosis\\u003c/b\\u003e \\u003c/p\\u003e \\u003cp\\u003eAO/EtBr dual staining is considered a very efficient method for the detection of early and late apoptotic cells. AO dye is permeable to all the cells and gives florescence in green color while EtBr is only permeable to the cells undergoing apoptosis and gives red fluorescence. In the dual staining, cells in the early apoptotic stage show a yellowish-orange color while cells in the late apoptotic stage show an orange-red color. We have found cells appeared as yellow-orange-red color cells in early and late apoptosis as shown in Fig.\\u0026nbsp;14.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec40\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eTo check anticancer activity via Wound healing/Cells Scratch assay\\u003c/h2\\u003e \\u003cp\\u003eIn the wound healing assay cells in the control were able to grow back and cover most of the scratch are up to 55% as shown in Fig.\\u0026nbsp;15. In the cells treated with alginate-sealed particles, the scratch area increased by 10\\u0026ndash;15% showing that cells can grow and migrate well after the treatment with particles.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eAnnexin V/ FITC and PI dual staining\\u003c/h3\\u003e\\n\\u003cp\\u003eIn the Annexin V/ FITC and PI dual staining, it was found that no significant death in the Untreated cells. Cells in the early stages of apoptosis show green fluorescence due to the Annexin V/FITC stain. Cells that are in the later stages of apoptosis or necrotic cells allow PI to enter into the cells due to the membrane disintegration and give red color fluorescence (Kumar et al., \\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). In this dual staining method, exposure with dummy β-Glucan particles leads to some of the cells in the apoptotic stage while quercetin-loaded particles are reported to have a significant number of cells in the later stage of apoptosis. Cells that were treated with quercetin loaded alginate sealed particles were reported to have the highest number of cells in the later stage of apoptosis or necrosis as visible in microscopic photographs Fig.\\u0026nbsp;16.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eIn this study, we have presented a comprehensive \\u003cem\\u003ein silico\\u003c/em\\u003e and experimental investigation into potential therapeutic agents for prostate cancer. The study utilized \\u003cem\\u003ein silico\\u003c/em\\u003e methods to identify potential drug targets associated with prostate cancer. By mining databases like DisGeNET and GeneCards, we identified a substantial number of genes linked to prostate cancer. This approach aligns with contemporary trends in bioinformatics-driven target identification, which has become increasingly crucial in precision medicine initiatives targeting cancer. A review summarizes the anticancer efficacy of quercetin against prostate cancer using PC3 and LNCaP cell lines. This study also highlighted that combination therapy of quercetin for prostate cancer treatment has various advantages such as enhanced anticancer effect, reduction in the treatment dose along with lower side effects (Yang et al., \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). The binding affinity of β-Glucan and quercetin with selected targets relevant to prostate cancer, namely CYP19ABG, ABCB1, and CDK1 was determined with the help of molecular docking. The observation of a higher binding affinity of quercetin compared to β-Glucan underscores its potential as a therapeutic agent. This finding corroborates recent studies highlighting the anti-cancer properties of quercetin through various mechanisms, including apoptosis induction and inhibition of cancer cell proliferation. A study demonstrated the apoptotic effect of quercetin against a panel of 9 cancer cell lines including prostate cancer and quercetin significantly induced apoptosis in these cancer cells (Hashemzaei et al., \\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eWe made formulation by loading quercetin into β-Glucan derived from Agaricus bisporus and yeast and alginate sealing was done for the slow and sustained release of quercetin from the hollow β-Glucan particles. The alginate-sealed formulation was reported to slow down the release of the compound in many studies (Soto et al., \\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). We conducted MTT assays on DU145 cells to evaluate the cytotoxic effects of particles derived from Agaricus bisporus and yeast, demonstrating a dose-dependent decrease in cell viability. Importantly, yeast-derived particles, particularly quercetin-loaded and alginate-sealed particles, exhibited promising cytotoxicity against prostate cancer cells. We found that the alginate-sealed particles i.e. ABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG and YBG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG have having lowest IC50 values so they can be considered good anticancer agents against prostate cancer. We also conducted MTT on the healthy cell line RAW264.7 to assess the effect of these particles and found that particles are not affecting the viability of the cells at initial concentration. At higher concentrations such as 500\\u0026micro;g/ml, cell viability was decreasing up to 20%. A study conducted the MTT of β-Glucan on RAW264.7 cell line and found that the control group and β-Glucan treated group did not have any significant difference in cell survival and this study aligns with our study in which we found no significant cell death after the treatment with β-Glucan and its formulations (Choi et al., \\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eThrough fluorescence microscopy and staining techniques, we tried to find out mechanistic insights into the mode of action of the particles. The observed increase in ROS generation, alterations in lysosomal pH, decline in MMP, and induction of apoptosis highlight the significant anti-cancer potential of these compounds. Quercetin is investigated in many studies and is found to attenuate cell survival, inhibit cell proliferation pathways, and induce cell death due to the generation of ROS (Ward et al., \\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e) and slow and sustained release of the Quercetin from the hollow β-Glucan particles due to alginate sealing will enhance its potential towards efficient anticancer activity. In this study, ROS generation increased in a dose-dependent manner that confirmed the apoptosis of the cancer cells due to oxidative stress. Nuclear fragmentation and condensation in DU145 after treatment with β-Glucan particles aligned with a study in which Farnesol was found to induce apoptosis in DU145 cells as assessed through a fluorescent microscope using DAPI staining (Park et al., \\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eAO/EtBr dual staining was found to show cells in the early and late apoptotic stages. AO stains live cells while EtBr stains the cells undergoing apoptosis or having compromised membranes. In a study, PAX2 siRNA knocked down DU145 cells and showed condensed yellow nuclei due to the co-localization of AO and EtBr dye (Bose et al., \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2009\\u003c/span\\u003e). Cell scratch assay confirmed that cells are losing their capability to migrate as was studied in a study in which cyclosporine A was able to inhibit the 25\\u0026ndash;33% motility of tumor cells in DU145 cells (Cevik et al., \\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). This can be very beneficial because the migration of cells from one place to another can result in the metastasis of prostate cancer. AnnexinV/FITC and PI dual staining showed the cells in the early apoptotic and apoptotic/necrotic stages. AnnexinV/FITC and PI dual staining determines apoptotic cells due to the alteration in the plasma membrane. The translocation of the phosphatidyl-serine from the inner to the outer membrane leads to its binding with Annexin V showing cells in early stages of apoptosis while later stages of apoptosis allow PI to permeate into the cells due to pore formation in the plasma membrane resulting in binding with Annexin V and PI both. Quercetin-loaded and alginate-sealed particles were reported to have a significant number of cells in the later stage of apoptosis showing the efficacy of the formulation (Yusof et al., 2019). Overall, our findings suggested that particles derived from Agaricus bisporus and yeast have efficient anticancer activity and further in vivo studies and clinical investigations can give impactful findings to use these formulations for the treatment of prostate cancer.\\u003c/p\\u003e\"},{\"header\":\"Conclusions\",\"content\":\"\\u003cp\\u003eThe integrated \\u003cem\\u003ein silico\\u003c/em\\u003e and \\u003cem\\u003ein vitro\\u003c/em\\u003e study underscores the potential of quercetin-loaded and alginate-sealed β-Glucan particles as promising therapeutic candidates for prostate cancer treatment. Through a combination of computational target identification and \\u003cem\\u003ein vitro\\u003c/em\\u003e experimental validation, the study elucidated the higher binding affinity of quercetin and cytotoxic effects of quercetin-loaded particles, particularly from yeast sources, against DU145 cells. ROS generation and other fluorescent assays support the anti-cancer properties of these particles. Future directions could involve preclinical studies to assess efficacy in animal models, pharmacokinetic profiling to determine bioavailability, and clinical trials to evaluate safety and efficacy in human patients. These efforts hold promise for advancing precision oncology and improving patient outcomes in prostate cancer treatment.\\u003c/p\\u003e\"},{\"header\":\"Abbreviations\",\"content\":\"\\u003cdiv class=\\\"DefinitionList\\\"\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eABG\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003e \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derivedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eABG\\u0026thinsp;+\\u0026thinsp;Q\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003e \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived quercetin loadedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eABG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003e \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e derived quercetin-loaded alginate-sealedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eYBG\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003eYeast derivedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eYBG\\u0026thinsp;+\\u0026thinsp;Q\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003eYeast derived quercetin loadedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eYBG\\u0026thinsp;+\\u0026thinsp;Q\\u0026thinsp;+\\u0026thinsp;ALG\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003eYeast derived quercetin-loaded alginate-sealedβ-Glucan\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eROS\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003eReactive oxygen species\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003eAO/EtBr\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003eAcridine orange/ethidium bromide\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv class=\\\"DefinitionListEntry\\\"\\u003e \\u003cdiv class=\\\"Term\\\"\\u003e\\u003cb\\u003ePI\\u003c/b\\u003e\\u003c/div\\u003e \\u003cdiv class=\\\"Description\\\"\\u003e \\u003cp\\u003ePropidium Iodide\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003c/div\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eEthics approval and consent to participate\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot Applicable.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot Applicable in this section.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and materials\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWill be available on request.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompeting interests\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare no known competing interests.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot Applicable\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor\\u0026rsquo;s contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eRT has written original draft and performed experiments. TKU conceptualized the ideas, provided resources and supervised the research. PKP provided support during research and language editing. All authors read and approved the final version of manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAuthors are thankful to the Dr. Geetika Madan Patel, Vice President and Chairperson, Research and Development Cell, Parul University for providing laboratory facilities to conduct the proposed research work.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAtasoy \\u0026Ouml;, Erbaş O (2020) Up to date of prostate cancer. D J Med Sci 6(2):092-102.\\u003c/li\\u003e\\n\\u003cli\\u003eKlusa D, Lohaus F, Furesi G, Rauner M, Bene\\u0026scaron;ov\\u0026aacute; M, Krause M, Kurth I, Peitzsch C (2021) Metastatic spread in prostate cancer patients influencing radiotherapy response. Front. Oncol 10:627379.\\u003c/li\\u003e\\n\\u003cli\\u003eHou Z, Huang S, Li Z (2021) Androgens in prostate cancer: A tale that never ends. Cancer Lett. 516:1-2.\\u003c/li\\u003e\\n\\u003cli\\u003eOduwole OO, Huhtaniemi IT, Misrahi M (2021) The roles of luteinizing hormone, follicle-stimulating hormone and testosterone in spermatogenesis and folliculogenesis revisited. Int. J. Mol. Sci. 22(23):12735.\\u003c/li\\u003e\\n\\u003cli\\u003eMorote J, Triquell M, Campistol M, Abascal-Junquera JM, Servian P, Trilla E (2022) Stratifying the initial prostate cancer suspicion to avoid magnetic resonance exams by sequencing men according to serum prostate-specific antigen, digital rectal examination and the prostate-specific antigen density based on digital rectal prostate volume category. BJUI Compass. 4 (3): 266-8. 2022.\\u003c/li\\u003e\\n\\u003cli\\u003eBoard PA (2023) Prostate Cancer Treatment (PDQ\\u0026reg;). InPDQ Cancer Information Summaries [Internet]. National Cancer Institute (US).\\u003c/li\\u003e\\n\\u003cli\\u003eKaur R, Sharma M, Ji D, Xu M, Agyei D (2019) Structural features, modification, and functionalities of beta-glucan. Fibers. 8(1):1.\\u003c/li\\u003e\\n\\u003cli\\u003eNakajima M, Tanaka N, Motouchi S, Kobayashi K, Shimizu H, Abe K, Hosoyamada N, Abara N, Morimoto N, Hiramoto N, Nakata R (2024) Beta-Glucanase superfamily identified by sequential, functional, and structural analyses. bioRxiv. 2024-02.\\u003c/li\\u003e\\n\\u003cli\\u003eKupetz M, Procopio S, Sacher B, Becker T (2015) Critical review of the methods of \\u0026beta;-glucan analysis and its significance in the beer filtration process. Eur Food Res Technol. 1:725-36.\\u003c/li\\u003e\\n\\u003cli\\u003eCaseiro C, Dias JN, de Andrade Fontes CM, Bule P ((2022)) From cancer therapy to winemaking: The molecular structure and applications of \\u0026beta;-glucans and \\u0026beta;-1, 3-glucanases. Int. J. Mol. Sci. 23(6):3156.\\u003c/li\\u003e\\n\\u003cli\\u003eSletmoen M, Stokke BT (2008) Higher order structure of (1, 3)‐\\u0026beta;‐D‐glucans and its influence on their biological activities and complexation abilities Biopolymers: Original Research on Biomolecules. 9(4):310-21.\\u003c/li\\u003e\\n\\u003cli\\u003eYang D, Wang T, Long M, Li P (2020) Quercetin: its main pharmacological activity and potential application in clinical medicine. Oxid Med Cell Longev. 2020.\\u003c/li\\u003e\\n\\u003cli\\u003eGhafouri-Fard S, Shabestari FA, Vaezi S, Abak A, Shoorei H, Karimi A, Taheri M, Basiri A (2021) Emerging impact of quercetin in the treatment of prostate cancer. Biomed. Pharmacother. 138:111548.\\u003c/li\\u003e\\n\\u003cli\\u003eNazemi Z, Sahraro M, Janmohammadi M, Nourbakhsh MS, Savoji H (2023) A review on tragacanth gum: A promising natural polysaccharide in drug delivery and cell therapy. Int. J. Biol. Macromol. 124343.\\u003c/li\\u003e\\n\\u003cli\\u003eNam JS, Sharma AR, Nguyen LT, Chakraborty C, Sharma G, Lee SS (2016) Application of bioactive quercetin in oncotherapy: from nutrition to nanomedicine. Molecules. 21(1):108.\\u003c/li\\u003e\\n\\u003cli\\u003eCao H, Wang D, Gao R, Li C, Feng Y, Chen L (2022) Therapeutic targets and signaling pathways of active components of QiLing decoction against castration-resistant prostate cancer based on network pharmacology. PeerJ. 10:e13481.\\u003c/li\\u003e\\n\\u003cli\\u003eSuresh NT, Ravindran VE, Krishnakumar U (2021) A computational framework to identify cross association between complex disorders by protein-protein interaction network analysis. Curr. Bioinform. 16(3):433-45.\\u003c/li\\u003e\\n\\u003cli\\u003eYang L, Hu Z, Zhu J, Liang Q, Zhou H, Li J, Fan X, Zhao Z, Pan H, Fei B (2020) Systematic elucidation of the mechanism of quercetin against gastric cancer via network pharmacology approach. Biomed Res. Int. 2020.\\u003c/li\\u003e\\n\\u003cli\\u003eUpadhyay TK, Trivedi R, Khan F, Al-Keridis LA, Pandey P, Sharangi AB, Alshammari N, Abdullah NM, Yadav DK, Saeed M (2022) In vitro elucidation of antioxidant, antiproliferative, and apoptotic potential of yeast-derived \\u0026beta;-1, 3-glucan particles against cervical cancer cells. Front. oncol. 12:942075.\\u003c/li\\u003e\\n\\u003cli\\u003eBhosale A, Trivedi R, Upadhyay TK, Gurjar D, Aghaz F, Khan F, Pandey P, Zeyaullah M, Alam MS, Al-Najjar MA, Siddiqui S (2022) Investigation on Antimicrobial, Antioxidant, and Anti-cancerous activity of Agaricus bisporus derived \\u0026beta;-Glucan particles against cervical cancer cell line. Cell. Mol. Biol. 68(9):150-9.\\u003c/li\\u003e\\n\\u003cli\\u003eUpadhyay TK, Fatima N, Sharma D, Saravanakumar V, Sharma R (2017) Preparation and characterization of beta-glucan particles containing a payload of nanoembedded rifabutin for enhanced targeted delivery to macrophages. EXCLI journal.16:210.\\u003c/li\\u003e\\n\\u003cli\\u003eVodnik VV, Mojić M, Stamenović U, Otoničar M, Ajdžanović V, Maksimović-Ivanić D, Mijatović S, Marković MM, Barudžija T, Filipović B, Milo\\u0026scaron;ević V (2021) Development of genistein-loaded gold nanoparticles and their antitumor potential against prostate cancer cell lines. Mater. sci. eng. 124:112078.\\u003c/li\\u003e\\n\\u003cli\\u003eKhan F, Pandey P, Jha NK, Jafri A, Khan I (2020) Antiproliferative effect of Moringa oleifera methanolic leaf extract by down-regulation of Notch signaling in DU145 prostate cancer cells. Gene Rep. 19:100619.\\u003c/li\\u003e\\n\\u003cli\\u003eZhu X, Chen X, Wang G, Lei D, Chen X, Lin K, Li M, Lin H, Li D, Zheng Q (2022) Picropodophyllin inhibits the proliferation of human prostate cancer du145 and lncap cells via ros production and pi3k/akt pathway inhibition. Biol. Pharm. Bull. 45(8):1027-35.\\u003c/li\\u003e\\n\\u003cli\\u003eSana SS, Kumbhakar DV, Pasha A, Pawar SC, Grace AN, Singh RP, Nguyen VH, Le QV, Peng W (2020) Crotalaria verrucosa leaf extract mediated synthesis of zinc oxide nanoparticles: assessment of antimicrobial and anticancer activity. Molecules. 25(21):4896.\\u003c/li\\u003e\\n\\u003cli\\u003eYang Q, Fang Y, Zhang C, Liu X, Wu Y, Zhang Y, Yang J, Yong K (2022) Exposure to zinc induces lysosomal-mitochondrial axis-mediated apoptosis in PK-15 cells. Ecotoxicol. Environ. Saf. 241:113716.\\u003c/li\\u003e\\n\\u003cli\\u003eNing L, Zhao W, Gao H, Wu Y (2020) Hesperidin induces anticancer effects on human prostate cancer cells via ROS-mediated necrosis like cell death. J. buon. 25(6):2629-34.\\u003c/li\\u003e\\n\\u003cli\\u003eFernandes NV, Yeganehjoo H, Katuru R, DeBose-Boyd RA, Morris LL, Michon R, Yu ZL, Mo H (2013) Geranylgeraniol suppresses the viability of human DU145 prostate carcinoma cells and the level of HMG CoA reductase. Exp Biol Med. 238(11):1265-74.\\u003c/li\\u003e\\n\\u003cli\\u003eBobadilla AV, Ar\\u0026eacute;valo J, Sarr\\u0026oacute; E, Byrne HM, Maini PK, Carraro T, Balocco S, Meseguer A, Alarc\\u0026oacute;n T (2019) In vitro cell migration quantification method for scratch assays. J. R. Soc. Interface. 16(151):20180709.\\u003c/li\\u003e\\n\\u003cli\\u003eMd Yusof EN, M. Latif MA, M. Tahir MI, Sakoff JA, Simone MI, Page AJ, Veerakumarasivam A, Tiekink ER, BSA Ravoof T (2019) o-Vanillin derived schiff bases and their organotin (IV) compounds: synthesis, structural characterisation, \\u003cem\\u003ein-silico\\u003c/em\\u003e studies and cytotoxicity. Int. J. Mol. Sci. 20(4):854.\\u003c/li\\u003e\\n\\u003cli\\u003eLin T, Liang C, Peng W, Qiu Y, Peng L (2020) Mechanisms of core Chinese herbs against colorectal cancer: a study based on data mining and network pharmacology. J. evid.-based complement. altern. 2020.\\u003c/li\\u003e\\n\\u003cli\\u003eAmeer K, Murtaza MA, Zainab S, Kim YM, Arshad MU, Pasha I, Abid M, Park MK. Polysaccharide-Based Films: Carriers of Active Substances and Controlled Release Characteristics. InPolysaccharide Based Films for Food Packaging: Fundamentals, Properties and Applications 2024 May 23 (pp. 379-400). Singapore: Springer Nature Singapore.\\u003c/li\\u003e\\n\\u003cli\\u003eSahoo BM, Banik BK, Borah P, Jain A (2022) Reactive oxygen species (ROS): key components in cancer therapies. Anti-Cancer Agents Med. Chem (Formerly Current Medicinal Chemistry-Anti-Cancer Agents). 22(2):215-22.\\u003c/li\\u003e\\n\\u003cli\\u003eKumar R, Saneja A, Panda AK (2021) An annexin V-FITC\\u0026mdash;propidium iodide-based method for detecting apoptosis in a non-small cell lung cancer cell line. Lung Cancer: Methods and Protocols. 213-23.\\u003c/li\\u003e\\n\\u003cli\\u003eYang F, Song L, Wang H, Wang J, Xu Z, Xing N (2015) Quercetin in prostate cancer: Chemotherapeutic and chemopreventive effects, mechanisms and clinical application potential. Oncol. Rep. 33(6):2659-68.\\u003c/li\\u003e\\n\\u003cli\\u003eHashemzaei M, Delarami Far A, Yari A, Heravi RE, Tabrizian K, Taghdisi SM, Sadegh SE, Tsarouhas K, Kouretas D, Tzanakakis G, Nikitovic D (2017) Anticancer and apoptosis‑inducing effects of quercetin in vitro and in vivo. Oncology reports. 38(2):819-28.\\u003c/li\\u003e\\n\\u003cli\\u003eSoto E, Kim YS, Lee J, Kornfeld H, Ostroff G (2010) Glucan particle encapsulated rifampicin for targeted delivery to macrophages. Polymers. 2(4):681-9.\\u003c/li\\u003e\\n\\u003cli\\u003eChoi EY, Lee SS, Hyeon JY, Choe SH, Keum BR, Lim JM, Park DC, Choi IS, Cho KK (2016) Effects of \\u0026beta;-glucan on the release of nitric oxide by macrophages stimulated with lipopolysaccharide. Asian-Australas J Anim Sci. 29(11):1664.\\u003c/li\\u003e\\n\\u003cli\\u003eWard AB, Mir H, Kapur N, Gales DN, Carriere PP, Singh S (2018) Quercetin inhibits prostate cancer by attenuating cell survival and inhibiting anti-apoptotic pathways. World J. Surg. Oncol. 16:1-2.\\u003c/li\\u003e\\n\\u003cli\\u003ePark JS, Kwon JK, Kim HR, Kim HJ, Kim BS, Jung JY (2014) Farnesol induces apoptosis of DU145 prostate cancer cells through the PI3K/Akt and MAPK pathways. Int. J. Mol. Med. 33(5):1169-76.\\u003c/li\\u003e\\n\\u003cli\\u003eBose SK, Gibson W, Bullard RS, Donald CD (2009) PAX2 oncogene negatively regulates the expression of the host defense peptide human beta defensin-1 in prostate cancer. Mol. Immunol. 46(6):1140-8.\\u003c/li\\u003e\\n\\u003cli\\u003eCevik O, Turut FA, Acidereli H, Yildirim S (2019) Cyclosporine-A induces apoptosis in human prostate cancer cells PC3 and DU145 via downregulation of COX-2 and upregulation of TGF\\u0026beta;. Turk Biyokim Derg. 44(1):47-54.\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"bulletin-of-the-national-research-centre\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"bnrc\",\"sideBox\":\"Learn more about [Bulletin of the National Research Centre](https://BNRC.springeropen.com)\",\"snPcode\":\"42269\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/42269/3\",\"title\":\"Bulletin of the National Research Centre\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Open\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"β-Glucan, DU145 prostate cancer, Quercetin, network pharmacology, Apoptosis\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4486275/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4486275/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003ch2\\u003eBackground\\u003c/h2\\u003e \\u003cp\\u003eProstate cancer remains a challenge in healthcare, being the second most common male cancer demanding innovative therapeutic approaches and treatment techniques. This study integrates \\u003cem\\u003ein silico\\u003c/em\\u003e and \\u003cem\\u003ein vitro\\u003c/em\\u003e methods for the investigation of the potential anticancer effects of quercetin-loaded and alginate-sealed β-Glucan particles derived from mushroom \\u003cem\\u003eAgaricus bisporus\\u003c/em\\u003e and yeast against the DU145 cell line.\\u003c/p\\u003e\\u003ch2\\u003eMethods\\u003c/h2\\u003e \\u003cp\\u003eProstate cancer-related genes were identified from DisGeNET and GeneCards databases, followed by target prioritization using Swiss Target Prediction software. Venny 2.1 was used for the determination of common targets between β-Glucan, Quercetin, and prostate cancer and PPI network was constructed using STRING database. CB dock online server was used for molecular docking and DU145, RAW264.7 cell lines were used for the determination of cytotoxicity against prostate cancer and healthy cells.\\u003c/p\\u003e\\u003ch2\\u003eResults\\u003c/h2\\u003e \\u003cp\\u003eMolecular docking revealed that quercetin has superior binding affinity compared to β-Glucan with selected prostate cancer-related targets. \\u003cem\\u003eIn vitro\\u003c/em\\u003e evaluation using MTT assays demonstrated the cytotoxic effects of quercetin-loaded and alginate-sealed particles against DU145 prostate cancer cells. Apoptosis induction, ROS generation, and lysosomal pH alterations underscore the potential of quercetin-loaded and alginate-sealed β-Glucan particles as promising therapeutic agents for prostate cancer.\\u003c/p\\u003e\\u003ch2\\u003eConclusions\\u003c/h2\\u003e \\u003cp\\u003eOur study showed systematic analyses of the effect of hollow β-Glucan particles, Quercetin, and Quercetin alginate sealed particles against DU145 cells and found that formulation exhibits superior anticancer activity against prostate cancer cell line. Quercetin-loaded alginate-sealed particles showed very little cytotoxicity against healthy cell line RAW264.7. Future studies focusing on preclinical validation, pharmacokinetic profiling, and clinical trials to assess translational potential and optimize therapeutic strategies can help get impactful findings.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Quercetin loaded and alginate sealed β-Glucan particles-based drug delivery system against DU145 a prostate cancer cell line: Integrating network pharmacology, molecular docking and in vitro studies\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-06-24 03:08:08\",\"doi\":\"10.21203/rs.3.rs-4486275/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewerAgreed\",\"content\":\"\",\"date\":\"2024-06-10T07:17:45+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-06-08T22:01:06+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-06-06T05:46:09+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Bulletin of the National Research Centre\",\"date\":\"2024-06-05T04:57:04+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"bulletin-of-the-national-research-centre\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"bnrc\",\"sideBox\":\"Learn more about [Bulletin of the National Research Centre](https://BNRC.springeropen.com)\",\"snPcode\":\"42269\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/42269/3\",\"title\":\"Bulletin of the National Research Centre\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Open\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"93d4cee7-ae45-4b97-8eb5-573dda47f332\",\"owner\":[],\"postedDate\":\"June 24th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-06-24T03:08:08+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-06-24 03:08:08\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4486275\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4486275\",\"identity\":\"rs-4486275\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}