A super-assembled synergistically nanoplatform AP@ZIF-8 Pt for hepatocarcinoma therapy

A super-assembled synergistically nanoplatform AP@ZIF-8 Pt for hepatocarcinoma therapy | 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 A super-assembled synergistically nanoplatform AP@ZIF-8 Pt for hepatocarcinoma therapy Zhenzhen Luo, Dunhuang Wang, Lie Lin, Rui Zhou, Yuanyuan Su, Zongkai Zhang, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5708497/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Intensive cancer treatment with nanoplatform is widely exploited in the clinic, the emerging nanomedicine offers an unparalleled opportunity for encapsulating potential antitumor drugs in a nano-carrier. Apoptin (AP)，a coding protein of VP3 gene, stem from the chicken anemia virus (CAV), can be activated in malignant cells selectively and prevents the dividing cancer cells from repairing their DNA lesions, thereby forcing them to undergo apoptosis. Herein, a three-step intelligent biodegradable drug delivery nanoplatform was designed. First, a hollow ZIF-8 was synthesized, embedded with platinum nanoparticle to form ZIF-8 Pt , and then loaded with AP, and lastly formed AP@ZIF-8 Pt , which possess pH-responsive drug release and cancer-targeted ability. As expected, both in vitro and in vivo experiment demonstrated that AP@ZIF-8 Pt performed treatment effects in hepatocarcinoma through relieving tumor-hypoxic microenvironment, inhibiting cell proliferation, and promoting cell apoptosis. Further ranscriptomic analysis showed that the specific mechanism of the AP@ZIF-8 Pt was thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, prion disease and PI3K-Akt signaling pathway. This work highlights a new strategy for liver cancer treatment and provides a reference for treating malignant tumors. AP@ZIF-8Pt tumor-hypoxic microenvironment apoptosis PI3K-Akt signaling pathway Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction Liver cancer, specifically hepatocellular carcinoma (HCC), is one of the most frequent malignancies causes cancer-related death[ 1 – 3 ]. Although immune checkpoint blockade-based immunotherapies and targeted therapy of HCC have achieved remarkable success in the field of HCC treatment, few of them induce durable responses and provide prominent survival benefits in patients because of the HCC physiological complexity[ 4 – 6 ]. To overcome it, effective strategies to optimizing HCC treatment are urgently needed[ 7 , 8 ], and there are two emerged strategies: finding new drugs according specific mechanisms and developing novel drug delivery system to give full play to the new drugs[ 9 ]. As for the new drugs finding, drug selectivity to cancer cells is the key issue of antitumor therapy[ 10 ]. A group of proteins, killing cancer cells specifcally without harming the normal cells, has attracted scientific interest[ 11 ]. Apoptin (AP) is one of these proteins. In the normal cells, AP becomes filamentous to aggregated and then degraded through proteasomes. While in cancer cells, AP induces apoptosis [ 12 – 14 ]. Our early study has proved that AP could induce apoptosis in hepatocarcinoma cells through targeting XPO1[ 15 ]. So we focused on AP as the potential HCC therapy. As for the novel drug delivery system developing, the emerging nanoplatform provides a new direction, which delivering the drugs and regulating tumor-hypoxic microenvironment synergistically, therefore achieving remarkable therapeutic effect[ 16 ]. Among them, zeolitic imidazolate framework-8 (ZIF-8) is a metal-organic frameworks (MOFs) serving as a promising candidate for its stability in aqueous environments and decomposability in acidic environments [ 17 – 19 ]. Moreover, ZIF-8 possesses good biocompatibility, can reduce immune reactions and increase drugs bioavailability in vivo[ 20 – 22 ]. Platinum nanoparticles (Pt) can catalyze H 2 O 2 (100 M-1 mM) to produce oxygen, thereby relieving tumor hypoxia in tumor tissues[ 23 , 24 ]. Herein, we design a fire-new nanoplatform, using ZIF-8 implanted with Pt(ZIF-8 Pt ), after AP loading, the final AP@ZIF-8 Pt was used to HCC therapy. Firstly, AP@ZIF-8 Pt had decomposability in the weakly acidic tumor microenvironment (TME), and then releases loaded AP. Secondly, the released AP can induce apoptosis in hepatocarcinoma cells. Thirdly, Pt works as a catalyst, trigger endogenous H 2 O 2 into O2, relieving hypoxia of TME and further improving the drug efficacy of AP. Expectedly, both in vitro and in vivo experiment results demonstrated that AP@ZIF-8 Pt possessed the extraordinary biocompatibility and enhanced therapeutic effect through inducing apoptosis, promoting DNA damage of hepatocarcinoma cells. Futher ranscriptomic analysis showed that the specific mechanism of the AP@ZIF-8 Pt was thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, prion disease and PI3K-Akt signaling pathway. Collectively, this work highlights a new strategy for HCC therapy and can be a reference for developing the advanced antitumor therapy. 2. Results and Discussion 2.1. AP@ZIF-8 Pt synthesis and characterization First, zinc nitrate and 2-methylimidazole were used to synthesize zeolitic imidazolate framework 8 (ZIF-8)[ 25 ]. Second, polyvinylpyrrolidone (PVP)-modified Pt nanoparticles was added in to synthesize ZIF-8 Pt [ 26 ]. Third, loaded AP to form AP@ZIF-8 Pt . The form of ZIF-8 was first scanned by scanning electron microscope (SEM) (Fig. 1 A), made sure the framework was correct. The particle size and morphology of AP@ZIF-8 Pt was scanned by SEM (Fig. 1 B) and transmission electron microscopy (TEM) ( Fig. 1 C ) , verified nanoparticles successful formation. The hydrated particle size of AP@ZIF-8 Pt was measured by dynamic light scattering (DLS), which was about 150 nm ( Fig. 1 D ) . Additionally, the X-ray diffraction (XRD) was performed to analyze the structure of AP@ZIF-8 Pt . The result showed that AP@ZIF-8 Pt XRD pattern was highly consistent with AP@ZIF-8 alone ( Fig. 1 E ) , and the peaks (111), (200), (220), and (310) at high angles of 30º–90º corresponding to that of Pt ( Fig. 1 F ) , suggesting that Pt were coated on ZIF-8 successfully. Further pH-responsive decomposition behavior of AP@ZIF-8 Pt was investigated, as showed in Fig. 1 G, when the pH decreased to 6.0, the accumulative release of AP was significantly increased, suggesting that AP@ZIF-8 Pt was easier to degrade in acidic tumor microenvironment. Furthermore, the surface potential of AP@ZIF-8 Pt was + 26.2 mV measured by Zeta potential ( Fig. 1 H ) . Finally, size and polydispersity index (PDI) of AP@ZIF-8 Pt proved the outstanding stability of AP@ZIF-8 Pt , which contained stability in phosphate-buffered saline (PBS) with 10% fetal bovine serum (FBS) for a week, suggesting that AP@ZIF-8 Pt could be used in in vivo potentially. 2.2. Cellular uptake and ROS generation of AP@ZIF-8 Pt To verify the cellular uptake ability of AP@ZIF-8 Pt in HCC cells, AP@ZIF-8 Pt was added to Hep3B cells for 6h. Confocal imaging showed that after 3h, part of AP@ZIF-8 Pt entered the cells and almost all AP@ZIF-8 Pt entered the cells after 6h (Fig. 2 A), consistently with the flow cytometry analysis for quantitative detection result (Fig. 2 B). Further field emission Transmission electron microscopy (FE-TEM) was applied to verify the real-time absorption of AP@ZIF-8 Pt to Huh7 cells. The versatility of FE-TEM allows detailed characterization of the endocytosis of nanoparticles. FE-TEM images showed much AP@ZIF-8 Pt entered in HCT-116 cells (marked by blue circle) (Fig. 2 D). The biocompatibility of AP@ZIF-8 Pt was evaluated by Cell Counting Kit-8 (CCK-8) assay, when the concentration of AP@ZIF-8 Pt was up to 120 µg/mL, the survival rates of both normal liver cells (LO2) and HCC cells (Hep3B and Hepg2) were more than 95%, suggesting that the superior security of AP@ZIF-8 Pt . The reactive oxygen species (ROS) generation ability of AP@ZIF-8 Pt was evaluated by DCFH-DA, an ROS probe to detect H 2 O 2 content and oxidative stress[ 27 ]. As showed in Fig. 2 E, the green dots had no significant difference in the control and ZIF-8 treatment group, because the endogenous ROS producing in live cells. When treated with ZIF-8 Pt , the green dots significantly increased, as proved in our previous study, Pt nanoparticles could catalyze H 2 O 2 to O 2 [26] . The results showed that ZIF-8 Pt presented significant concentration-dependent catalase-like activity. 2.4. Therapeutic efficacy and mechanism of AP@ZIF-8 Pt CCK-8 assay was first used to verify therapeutic efficacy of AP@ZIF-8 Pt against three HCC cell lines (Hepg2, Hep3B and Huh7). The results showed that treatment with AP@ZIF-8 Pt significantly inhibits the viability of HCC cells compared with single AP group (Fig. 3 A). Flow cytometry was then used to verify therapeutic efficacy of AP@ZIF-8 Pt . The results showed that the apoptotic rate of AP@ZIF-8 Pt treatment group was up to 33.42% for 6h higher than 3h (15.48%) (Fig. 3 B, 4 C). Further therapeutic efficacy of AP@ZIF-8 Pt was determined in vivo. Hepg2 tumor-bearing mice were randomly divided into control group, ZIF-8 Pt group, AP group and AP@ZIF-8 Pt group. The body weights, tumor volumes and tumor weights of the mice were recorded every 2 days in order to assess the effects of different treatments. The body weights of mice increased steadily, demonstrating the biosafety of AP@ZIF-8 Pt (Fig. 3 D). The tumor weight and tumor volumes of mice treated with AP were smaller than those of the control group and ZIF-8 Pt group, and the smallest tumors were detected in AP@ZIF-8 Pt mice. (Fig. 3 E, F). Moreover, H&E and TUNEL staining analyses were performed to confirm the therapeutic efficacy of AP@ZIF-8 Pt . The necrosis and green dots detected in the AP are more than those observed in the control group and ZIF-8 Pt group, and the maximum necrosis and green dots were detected in AP@ZIF-8 Pt group, indicates the remarkable therapeutic efficacy of AP@ZIF-8 Pt (Fig. 3 G). The mechanism underlying the efficacy of AP@ZIF-8 Pt was determined by immunofluorescence staining. The results showed that AP@ZIF-8 Pt treatment up-regulated the expression of γ-H2AX, the DNA DSB marker[ 28 ] (Fig. 3 H), indicated that AP@ZIF-8 Pt promoted DNA damage by increasing the number of DSBs. In contrast, AP@ZIF-8 Pt treatment down-regulated the expression of HIF-1α, a regulator of primary adaptive responses to hypoxia [ 29 , 30 ] (Fig. 3 H), indicated that AP@ZIF-8 Pt could alleviate hypoxia in tumor tissue to improve therapy sensitivity. 2.5. Transcriptomics study Transcriptomics is an extremely important tool for studying all RNA molecules in an organism[ 31 ]. Cancer-associated protein expression can change levels of some RNA to promote cancer initiation and progression[ 32 ]. After confirming the in vivo anticancer efficacy of AP@ZIF-8 Pt , we further investigated the transcriptomics influence of it in Hepg2-tumor-bearing mice. The serum samples from mice treated with PBS, ZIF-8 Pt and AP@ZIF-8 Pt were used for transcriptomics analysis. The sample density result was shown in Fig. 4 A. All of the samples in each group were closely clustered, which demonstrated a good quality of samples with different groups. To focus on the differential genes influenced by AP@ZIF-8 Pt , we performed analysis of differential genes between two of each of the groups (control and ZIF-8 Pt , control and AP@ZIF-8 Pt , ZIF-8 Pt and AP@ZIF-8 Pt ) (Fig. 4 B). Differential transcriptomics analysis between the control and ZIF-8 Pt groups and between the control and AP@ZIF-8 Pt groups was carried out to eliminate the batch effect. A total of 173 genes were identified between the control and ZIF-8 Pt groups, 2201 genes were identified between the control and AP@ZIF-8 Pt groups, 1223 genes were identified between the ZIF-8 Pt and AP@ZIF-8 Pt groups. Finally, 1188 genes were identified as significant genes influenced by AP@ZIF-8 Pt (Fig. 4 C). The enrichment analysis was then performed for these significant genes. As showed in Fig. 4 D, the top 5 important Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathways were thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, rion disease and PI3K-Akt signaling pathway. As showed in Fig. 4 E, the top 3 important biological processes were purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process and ribonucleoside diphosphate metabolic process. The top 3 important cellular components were respiratory chain, cytosolic large ribosomal subunit and respiratory chain complex. And the 2 important molecular functions were extracellular mata structin consultent and Oxygen binding. Albert S Peixoto et al . proved that HCC induced by hepatocyte Pten deletion reduced thermogenic capacity[ 33 ]. As is known to all, the dysregulation of the PI3K-AKT-mTOR signaling pathway is common in HCC[ 34 ]. Meilin Xue et al . proved METTL16 promoted liver cancer stem cell self-renewal via controlling ribosome biogenesis and mRNA translation[ 35 ]. R Read et al . demonstrated theat Soluble enzyme hydrolyzes purine nucleoside diphosphate resulted in HCC[ 36 ]. 2.6. Biocompatibility of AP@ZIF-8 Pt It’s essential to evaluate the biosafety to determine whether AP@ZIF-8 Pt can be applied in clinical treatment[ 37 ]. Considering that changes in the behavior and body weights of mice are considered signs of acute toxicity and that changes in gross pathology reflect long-term toxicity [ 38 ], these parameters were assessed throughout the treatment period. Blood samples were collected from mice administered with i.p. injections of PBS, ZIF-8 Pt and AP@ZIF-8 Pt , and the behavior and body weights of these mice were monitored. The results showed that treatment with ZIF-8 Pt and AP@ZIF-8 Pt did not induce significant variations in mouse behavior and body weight compared to the control group (Fig. 3 D), which proved that the acute toxicity of ZIF-8 Pt and AP@ZIF-8 Pt was negligible. Similarly, routine blood tests proved that the ZIF-8 Pt and AP@ZIF-8 Pt groups did not cause hemolysis or myelosuppression even after 30 days of treatment (Fig. 4 A). Further liver and kidney function tests were performed to evaluate the liver or kidney damage induced by ZIF-8 Pt and AP@ZIF-8 Pt . ALT and AST were assessed as liver damage indexes, and BUN and CR were assessed as kidney damage indexes [ 39 – 41 ]. The results demonstrated that both ZIF-8 Pt and AP@ZIF-8 Pt did not damage liver and kidney functions (Fig. 4 B). H&E staining analysis proved that no organ damage was induced by both ZIF-8 Pt and AP@ZIF-8 Pt (Fig. 4 C). All of these results proved that both ZIF-8 Pt and AP@ZIF-8 Pt possessed highly biocompatible and potential of clinical transformation. 3. Conclusion In this study, a pH-responsive and catalase-like Pt nanoparticle-embedded metal-organic framework (ZIF-8 Pt ) loaded with AP (AP@ZIF-8 Pt ) was developed to treating HCC. Both in vitro and in vivo experiment results proved that AP@ZIF-8 Pt had excellent biocompatibility and therapeutic efficacy. Further experiment verified that AP@ZIF-8 Pt can promote DNA damage and relieving hypoxia. Transcriptomics analysis demonstrated that AP@ZIF-8 Pt worked through thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, rion disease, PI3K-Akt signaling pathway, purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process and ribonucleoside diphosphate metabolic process, respiratory chain, cytosolic large ribosomal subunit, respiratory chain complex, extracellular mata structin consultent and Oxygen binding. Our study suggested that this nanoplatform AP@ZIF-8 Pt offered a promising paradigm for HCC treatment and had clinical application potential. 4. Materials and Methods 4.1. Materials 1,2-dimethylimidazole (2-MIL), zinc nitrate hexahydrate (Zn (NO 3 ) 2 ·6H 2 O), chloroplatinic acid hydrate (H 2 PtCl 6 ·6H 2 O) and polyvinylpyrrolidone (PVP) were purchased from Aladdin. AP was synthesized by Shanghai Qiangyao Biotechnology Co., Ltd. 4'6-diamidino-2-phenylindole (DAPI) and 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Beyotime Biotechnology. The γ-H2AX antibody, HIF-1α antibody and secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). All of the reagents used in the experiments were of analytical grade and used directly without treatment. 4.2. Characterizations Malvern Zetasizer Nano ZS90 Apparatus, UV-2550 Ultraviolet-visible Spectrophotometer (Shimadzu, Japan), transmission electron microscopy system (JEOL/JEM-F200), XRD (Bruker/D8 ADVANCE), FTIR (Bruker/VERTEX70), FC500 flow cytometer (Beckman Coulter, USA) were used for material characterization same as our previous studies[26, 42]. 4. 3. Synthesis of AP@ZIF-8 Pt Zinc nitrate hexahydrate (25 mM) and 2-methyl imidazole (200 mM) were first dissolved in methanol. Then, 20 mL of methanol solution was stirred together with Pt nanoparticles. After continuous stirring for 3–4 h, the sample was washed three times with methanol, and ZIF-8 Pt was obtained. Then redispersed ZIF-8 Pt in ethanol (10 mL) added AP and stirred for 6–8 h. The mixed solution was then washed with methanol (10000 rpm, 10 min) to remove excess AP then obtained AP@ZIF-8 Pt . 4. 4. In vitro cytotoxicity assay The cytotoxicity against HCC cells was measured by CCK8 assay. The cells were cultured in a 96-well plate and incubated overnight. Next, gradient concentrations of AP@ZIF-8 Pt (0, 20, 40, 60, 80 and 100 μg/mL) were added to each well and incubated for 24 h. Five parallel samples were set for each group, then 10 µL of CCK8 was added to each well and incubated for 4 h. The absorbance was quantified by a spectrophotometer at 450 nm. 4. 5. R e active oxygen species (ROS) detection ROS in cells was detected using the ROS probe, DCFH-DA. Briefly, HCC cells were seeded in a 6-well plate and cultured overnight. ZIF-8 and ZIF-8 Pt were added to the cells and incubated in the dark for 2 h. Following incubation, the medium was removed and cells were washed with PBS three times. Following washing, the ROS probe, DCFH-DA, was added to the cells and incubated for a further 30 min, and then the fluorescence of cells generated by DCFH-DA was detected by fluorescence microscopy. 4. 6. Cellular uptake analysis HCC cells were seeded in confocal chambers and cultured overnight. The AP loaded Rhodamine B to track nanoparticles, were added to the chambers and incubated for 2 h. Next, the cells were washed with PBS three times and stained with phalloidin-FITC (30 min) and DAPI (10 min). Finally, the cells were observed by laser confocal microscopy (phalloidin-FITC, excitation: 490 nm; DAPI, excitation: 400 nm, and Rhodamine B, excitation: 555 nm). We further investigated the cellular uptake of nanoparticles by FE-TEM. Briefly, HCC cells were cultured in a cell slide overnight. After incubating with AP@ZIF-8 Pt for 2 h, the cell slides were washed with PBS. The cells were fixed with 3% glutaraldehyde overnight and then dehydrated in an ethanol series. Using a mini-gold sputter, cell samples were coated with gold-palladium for visual inspection by TEM. 4. 7. Flow cytometry analysis HCC cells at a concentration of 1 × 10 5 cells were seeded in six-well plates and incubated overnight. Cells were treated with AP@ZIF-8 Pt , which were loaded with rose red B. After incubation for 3 h and 6h, the cells were washed three times with cold PBS and the treated cells were harvested for flow cytometry analysis.The cells were seeded in 6 well plates at a density of 2×10 5 cells and cultured overnight. Apoptosis kit (BD Biosciences, CA) was used for apoptosis analysis and operated according to the instruction. 4. 8. Immunofluorescence HCC cells were fixed with 4% PFA at 37°C for 20 min, treated with 0.02% Triton X-100 for 10 min, and then blocked at room temperature for 1 h. Cells were incubated with primary antibody overnight at 4°C followed by incubation with fluorescent-conjugated secondary antibody for 2 h at room temperature. The nuclei were stained with DAPI for 15 min. The images of the cells were obtained by laser confocal microscopy (γ-H2AX-FITC, excitation: 490 nm, HIF-1α-CY5, excitation: 650 nm and DAPI, excitation: 400 nm). 4. 9. In vivo therapeutic effect When the tumor reached approximately 100 mm 3 , tumor-bearing mice were randomly divided into 4 groups: (I) Control; (II) AP; (III) ZIF-8 Pt , and (IV) AP@ZIF-8 Pt . After the nanoparticles (50 mg∙kg −1 ) were intravenously injected into the tail veins every 2 d. For treatment, the solid tumor was dissected from one mouse in each group for H&E and TUNEL staining. The solid tumor was also used for HIF-1α and γ-H2AX IHC staining. The body weights of mice and the tumor volumes were recorded every other day. 4. 10. Transcriptomics analysis When the tumor reached about100 mm 3 , tumor-bearing mice were randomly divided into three groups: (I) Control; (II) ZIF-8 Pt ; (III) AP@ZIF-8 Pt . Nanoparticles (50 mg/kg) were intravenously injected via tail vein every 2d. At the end of the experiment, tumors were dissected for transcriptomics analysis using an LC-ESI-MS/MS system. 4. 16. Statistical analyses One-way and two-way analyses of variance (ANOVA) and t-tests were used to determine statistical significance (*p<0.05, **p<0.01, ***p<0.001), p-value less than 0.05 was considered statistically significant. Declarations Supporting Information Supporting Information is available from the Wiley Online Library or from the author. Funding Declaration This work was supported by the National Natural Science Foundation of China (No.82403783), the Nature Science Foundation of Fujian Province (No.2024J08316) and National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital & Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen (No.E010322014) . Declaration of Interest Statement The authors declare no competing financial interest. Ethics and Consent to Participate declarations All animal experiments were approved by the Experimental Animal Management and Ethics Committee of Xiamen University. Author Contribution Zhenzhen Luo and Dunhuang Wang wrote the main manuscript text; Lie Lin and Rui Zhou prepared figures 1-2. Yuanyuan Su, Zongkai Zhang and Jing Hu prepared figures 3-4; Yaqing Dai and Xiaoyan Huang prepared figure 5; Yufei Zhou and Liuyun Gong reviewed the manuscript. 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Wang, J., et al., One Stone, Two Birds: A Peptide-Au(I) Infinite Coordination Supermolecule for the Confederate Physical and Biological Radiosensitization in Cancer Radiation Therapy. Small, 2023. 19 (11): p. e2204238. Additional Declarations No competing interests reported. Supplementary Files floatimage1.png Graphic Abstract: A nanoplatform using ZIF-8 implanted with Pt(ZIF-8 Pt ), after AP loading, the final AP@ZIF-8 Pt was used to HCC therapy. Firstly, AP@ZIF-8 Pt had decomposability in the weakly acidic tumor microenvironment (TME), and then releases loaded AP. Secondly, the released AP can induce apoptosis in hepatocarcinoma cells. Thirdly, Pt works as a catalyst, trigger endogenous H 2 O 2 into O 2 , relieving hypoxia of TME and further improving the drug efficacy of AP. 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A. \u003c/strong\u003eScanning electron microscope (SEM) image of ZIF-8. \u003cstrong\u003eB. \u003c/strong\u003eSEM image of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. \u003cstrong\u003eC.\u003c/strong\u003e Transmission electron microscopy (TEM) image of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. \u003cstrong\u003eD.\u003c/strong\u003e Hydrated particle size of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. \u003cstrong\u003eE.\u003c/strong\u003e XRD patterns of AP@ZIF-8 and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. \u003cstrong\u003eF. \u003c/strong\u003eXRD patterns in Pt region. \u003cstrong\u003eG\u003c/strong\u003e. The cumulative release profile of AP (n=5). \u003cstrong\u003eH\u003c/strong\u003e. Zeta potential of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e\u003cstrong\u003e. I. \u003c/strong\u003eChanges of diameter and PDI of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e over one week.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/e24958924d55d3fc99904c4e.png"},{"id":72790466,"identity":"4eae8493-d109-431b-9a0f-48abd6d54901","added_by":"auto","created_at":"2025-01-02 08:09:09","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":642229,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCellular uptake and ROS generation of AP@ZIF-8\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003ePt\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e A. \u003c/strong\u003eAP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e incubated with HCC cells for 3h and 6h. \u003cstrong\u003eB. \u003c/strong\u003eFlow cytometry analysis for quantitative detection of intracellular AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e fluorescence. \u003cstrong\u003eC. \u003c/strong\u003eDetailed characterization of endocytosis by FE-TEM observation. \u003cstrong\u003eD. \u003c/strong\u003eCell viability of LO2, Hep3B and Hepg2 cells with AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e treatment were evaluated by CCK-8 assay (n=5). E. ROS production detected by fluorescence of DCFH-DA in HCC cells.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/50b3334d001128dbc01302f8.png"},{"id":72790462,"identity":"2fb5849e-8a35-4b64-a776-0bc0a6dc1c9a","added_by":"auto","created_at":"2025-01-02 08:09:09","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":672168,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTherapeutic efficacy and mechanism of AP@ZIF-8\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003ePt\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e. A. \u003c/strong\u003eCell Counting Kit-8 (CCK8) assay results. \u003cstrong\u003eB.\u003c/strong\u003e Apoptosis analysis results. \u003cstrong\u003eC.\u003c/strong\u003e Quantification results of B, n=3. \u003cstrong\u003eD.\u003c/strong\u003e Body weights profiles, (n=5). \u003cstrong\u003eE. \u003c/strong\u003eTumor weights in each group were measured at the end of the experiment (n=5). \u003cstrong\u003eF.\u003c/strong\u003e Tumor growth profiles (n=5). \u003cstrong\u003eG.\u003c/strong\u003e Histologic analysis and TUNEL staining of tumor sections from different groups of mice (n=5). \u003cstrong\u003eH.\u003c/strong\u003e Immunofluorescence analysis of γ-H2AX and HIF-1α in tumor tissues collected from various groups of mice (n=5).\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/bebec4aad405f1c70c4a6c73.png"},{"id":72790456,"identity":"17a4f611-5704-4070-a30f-8f939c58346e","added_by":"auto","created_at":"2025-01-02 08:09:09","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":590797,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTranscriptomics analysis. A.\u003c/strong\u003e Sample density distribution. \u003cstrong\u003eB.\u003c/strong\u003e Volcano map of differential genes in each treatment group. \u003cstrong\u003eC.\u003c/strong\u003e Venn diagrams of significant differential genes influenced by each treatment group. \u003cstrong\u003eD.\u003c/strong\u003e Pathway enrichment analysis of significant genes influenced by AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. \u003cstrong\u003eE.\u003c/strong\u003e Gene ontology (GO) enrichment analysis of significant genes influenced by AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e.\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/a6190c57737c7308e274dba6.png"},{"id":72790449,"identity":"d2156c3d-9f0e-4b72-8788-25e2f47808dc","added_by":"auto","created_at":"2025-01-02 08:09:08","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":700405,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eBiocompatibility of AP@ZIF-8Pt\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003ein vivo. A.\u003c/strong\u003e Changes of routine blood test contained hemoglobin (HGB), platelet (PLT), red blood cell (RBC) and white blood cell (WBC). \u003cstrong\u003eB.\u003c/strong\u003e Changes of hepatorenal function contained alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN) and creatinine (CR). \u003cstrong\u003eC. \u003c/strong\u003eHE staining of mice after three injections of PBS, ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e.\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/ab71d82aca8e54f3143463ed.png"},{"id":72838677,"identity":"9022d16c-f702-4aaa-b277-91a47b5a166d","added_by":"auto","created_at":"2025-01-02 17:16:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3603977,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/4cdf633e-82d5-4105-b6d3-2f7fdcfa2870.pdf"},{"id":72790732,"identity":"22662926-35da-4bd4-ab42-cfc303dd0d6e","added_by":"auto","created_at":"2025-01-02 08:17:08","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":356385,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphic Abstract\u003c/strong\u003e: A nanoplatform using ZIF-8 implanted with Pt(ZIF-8\u003csup\u003ePt\u003c/sup\u003e), after AP loading, the final AP@ZIF-8\u003csup\u003ePt \u003c/sup\u003ewas used to HCC therapy. Firstly, AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e had decomposability in the weakly acidic tumor microenvironment (TME), and then releases loaded AP. Secondly, the released AP can induce apoptosis in hepatocarcinoma cells. Thirdly, Pt works as a catalyst, trigger endogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e into O\u003csub\u003e2\u003c/sub\u003e, relieving hypoxia of TME and further improving the drug efficacy of AP.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-5708497/v1/e812c4dc5bc475c392cb4012.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"A super-assembled synergistically nanoplatform AP@ZIF-8 Pt for hepatocarcinoma therapy","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eLiver cancer, specifically hepatocellular carcinoma (HCC), is one of the most frequent malignancies causes cancer-related death[\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Although immune checkpoint blockade-based immunotherapies and targeted therapy of HCC have achieved remarkable success in the field of HCC treatment, few of them induce durable responses and provide prominent survival benefits in patients because of the HCC physiological complexity[\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. To overcome it, effective strategies to optimizing HCC treatment are urgently needed[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], and there are two emerged strategies: finding new drugs according specific mechanisms and developing novel drug delivery system to give full play to the new drugs[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs for the new drugs finding, drug selectivity to cancer cells is the key issue of antitumor therapy[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. A group of proteins, killing cancer cells specifcally without harming the normal cells, has attracted scientific interest[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Apoptin (AP) is one of these proteins. In the normal cells, AP becomes filamentous to aggregated and then degraded through proteasomes. While in cancer cells, AP induces apoptosis [\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Our early study has proved that AP could induce apoptosis in hepatocarcinoma cells through targeting XPO1[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. So we focused on AP as the potential HCC therapy. As for the novel drug delivery system developing, the emerging nanoplatform provides a new direction, which delivering the drugs and regulating tumor-hypoxic microenvironment synergistically, therefore achieving remarkable therapeutic effect[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Among them, zeolitic imidazolate framework-8 (ZIF-8) is a metal-organic frameworks (MOFs) serving as a promising candidate for its stability in aqueous environments and decomposability in acidic environments [\u003cspan additionalcitationids=\"CR18\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Moreover, ZIF-8 possesses good biocompatibility, can reduce immune reactions and increase drugs bioavailability in vivo[\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Platinum nanoparticles (Pt) can catalyze H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (100 M-1 mM) to produce oxygen, thereby relieving tumor hypoxia in tumor tissues[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHerein, we design a fire-new nanoplatform, using ZIF-8 implanted with Pt(ZIF-8\u003csup\u003ePt\u003c/sup\u003e), after AP loading, the final AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was used to HCC therapy. Firstly, AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e had decomposability in the weakly acidic tumor microenvironment (TME), and then releases loaded AP. Secondly, the released AP can induce apoptosis in hepatocarcinoma cells. Thirdly, Pt works as a catalyst, trigger endogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e into O2, relieving hypoxia of TME and further improving the drug efficacy of AP. Expectedly, both in vitro and in vivo experiment results demonstrated that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e possessed the extraordinary biocompatibility and enhanced therapeutic effect through inducing apoptosis, promoting DNA damage of hepatocarcinoma cells. Futher ranscriptomic analysis showed that the specific mechanism of the AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, prion disease and PI3K-Akt signaling pathway. Collectively, this work highlights a new strategy for HCC therapy and can be a reference for developing the advanced antitumor therapy.\u003c/p\u003e"},{"header":"2. Results and Discussion","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e \u003cem\u003esynthesis and characterization\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eFirst, zinc nitrate and 2-methylimidazole were used to synthesize zeolitic imidazolate framework 8 (ZIF-8)[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Second, polyvinylpyrrolidone (PVP)-modified Pt nanoparticles was added in to synthesize ZIF-8\u003csup\u003ePt\u003c/sup\u003e[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Third, loaded AP to form AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. The form of ZIF-8 was first scanned by scanning electron microscope (SEM) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA), made sure the framework was correct. The particle size and morphology of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was scanned by SEM (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB) and transmission electron microscopy (TEM) \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC\u003cb\u003e)\u003c/b\u003e, verified nanoparticles successful formation. The hydrated particle size of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was measured by dynamic light scattering (DLS), which was about 150 nm \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD\u003cb\u003e)\u003c/b\u003e. Additionally, the X-ray diffraction (XRD) was performed to analyze the structure of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. The result showed that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e XRD pattern was highly consistent with AP@ZIF-8 alone \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eE\u003cb\u003e)\u003c/b\u003e, and the peaks (111), (200), (220), and (310) at high angles of 30\u0026ordm;\u0026ndash;90\u0026ordm; corresponding to that of Pt \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eF\u003cb\u003e)\u003c/b\u003e, suggesting that Pt were coated on ZIF-8 successfully. Further pH-responsive decomposition behavior of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was investigated, as showed in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eG, when the pH decreased to 6.0, the accumulative release of AP was significantly increased, suggesting that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was easier to degrade in acidic tumor microenvironment. Furthermore, the surface potential of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was +\u0026thinsp;26.2 mV measured by Zeta potential \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eH\u003cb\u003e)\u003c/b\u003e. Finally, size and polydispersity index (PDI) of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e proved the outstanding stability of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, which contained stability in phosphate-buffered saline (PBS) with 10% fetal bovine serum (FBS) for a week, suggesting that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e could be used in \u003cem\u003ein vivo\u003c/em\u003e potentially.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. \u003cem\u003eCellular uptake and ROS generation of AP@ZIF-8\u003c/em\u003e\u003csup\u003e\u003cem\u003ePt\u003c/em\u003e\u003c/sup\u003e\u003c/h2\u003e \u003cp\u003eTo verify the cellular uptake ability of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e in HCC cells, AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was added to Hep3B cells for 6h. Confocal imaging showed that after 3h, part of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e entered the cells and almost all AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e entered the cells after 6h (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA), consistently with the flow cytometry analysis for quantitative detection result (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Further field emission Transmission electron microscopy (FE-TEM) was applied to verify the real-time absorption of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e to Huh7 cells. The versatility of FE-TEM allows detailed characterization of the endocytosis of nanoparticles. FE-TEM images showed much AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e entered in HCT-116 cells (marked by blue circle) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD). The biocompatibility of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was evaluated by Cell Counting Kit-8 (CCK-8) assay, when the concentration of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was up to 120 \u0026micro;g/mL, the survival rates of both normal liver cells (LO2) and HCC cells (Hep3B and Hepg2) were more than 95%, suggesting that the superior security of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. The reactive oxygen species (ROS) generation ability of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was evaluated by DCFH-DA, an ROS probe to detect H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e content and oxidative stress[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. As showed in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE, the green dots had no significant difference in the control and ZIF-8 treatment group, because the endogenous ROS producing in live cells. When treated with ZIF-8\u003csup\u003ePt\u003c/sup\u003e, the green dots significantly increased, as proved in our previous study, Pt nanoparticles could catalyze H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e to O\u003csub\u003e2 [26]\u003c/sub\u003e. The results showed that ZIF-8\u003csup\u003ePt\u003c/sup\u003e presented significant concentration-dependent catalase-like activity.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.4. \u003cem\u003eTherapeutic efficacy and mechanism of\u003c/em\u003e AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e\u003c/h2\u003e \u003cp\u003eCCK-8 assay was first used to verify therapeutic efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e against three HCC cell lines (Hepg2, Hep3B and Huh7). The results showed that treatment with AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e significantly inhibits the viability of HCC cells compared with single AP group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). Flow cytometry was then used to verify therapeutic efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. The results showed that the apoptotic rate of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e treatment group was up to 33.42% for 6h higher than 3h (15.48%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC). Further therapeutic efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was determined in vivo. Hepg2 tumor-bearing mice were randomly divided into control group, ZIF-8\u003csup\u003ePt\u003c/sup\u003e group, AP group and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e group. The body weights, tumor volumes and tumor weights of the mice were recorded every 2 days in order to assess the effects of different treatments. The body weights of mice increased steadily, demonstrating the biosafety of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). The tumor weight and tumor volumes of mice treated with AP were smaller than those of the control group and ZIF-8\u003csup\u003ePt\u003c/sup\u003e group, and the smallest tumors were detected in AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e mice. (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE, F).\u003c/p\u003e \u003cp\u003eMoreover, H\u0026amp;E and TUNEL staining analyses were performed to confirm the therapeutic efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. The necrosis and green dots detected in the AP are more than those observed in the control group and ZIF-8\u003csup\u003ePt\u003c/sup\u003e group, and the maximum necrosis and green dots were detected in AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e group, indicates the remarkable therapeutic efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eG). The mechanism underlying the efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was determined by immunofluorescence staining. The results showed that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e treatment up-regulated the expression of γ-H2AX, the DNA DSB marker[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eH), indicated that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e promoted DNA damage by increasing the number of DSBs. In contrast, AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e treatment down-regulated the expression of HIF-1α, a regulator of primary adaptive responses to hypoxia [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eH), indicated that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e could alleviate hypoxia in tumor tissue to improve therapy sensitivity.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.5. \u003cem\u003eTranscriptomics study\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eTranscriptomics is an extremely important tool for studying all RNA molecules in an organism[\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Cancer-associated protein expression can change levels of some RNA to promote cancer initiation and progression[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. After confirming the in vivo anticancer efficacy of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, we further investigated the transcriptomics influence of it in Hepg2-tumor-bearing mice. The serum samples from mice treated with PBS, ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e were used for transcriptomics analysis. The sample density result was shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA. All of the samples in each group were closely clustered, which demonstrated a good quality of samples with different groups. To focus on the differential genes influenced by AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, we performed analysis of differential genes between two of each of the groups (control and ZIF-8\u003csup\u003ePt\u003c/sup\u003e, control and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB). Differential transcriptomics analysis between the control and ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups and between the control and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups was carried out to eliminate the batch effect. A total of 173 genes were identified between the control and ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups, 2201 genes were identified between the control and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups, 1223 genes were identified between the ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups. Finally, 1188 genes were identified as significant genes influenced by AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eThe enrichment analysis was then performed for these significant genes. As showed in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD, the top 5 important Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathways were thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, rion disease and PI3K-Akt signaling pathway. As showed in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE, the top 3 important biological processes were purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process and ribonucleoside diphosphate metabolic process. The top 3 important cellular components were respiratory chain, cytosolic large ribosomal subunit and respiratory chain complex. And the 2 important molecular functions were extracellular mata structin consultent and Oxygen binding. Albert S Peixoto \u003cem\u003eet al\u003c/em\u003e. proved that HCC induced by hepatocyte Pten deletion reduced thermogenic capacity[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. As is known to all, the dysregulation of the PI3K-AKT-mTOR signaling pathway is common in HCC[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Meilin Xue \u003cem\u003eet al\u003c/em\u003e. proved METTL16 promoted liver cancer stem cell self-renewal via controlling ribosome biogenesis and mRNA translation[\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. R Read \u003cem\u003eet al\u003c/em\u003e. demonstrated theat Soluble enzyme hydrolyzes purine nucleoside diphosphate resulted in HCC[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.6.\u003cem\u003eBiocompatibility of AP@ZIF-8\u003c/em\u003e\u003csup\u003e\u003cem\u003ePt\u003c/em\u003e\u003c/sup\u003e\u003c/h2\u003e \u003cp\u003eIt\u0026rsquo;s essential to evaluate the biosafety to determine whether AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e can be applied in clinical treatment[\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Considering that changes in the behavior and body weights of mice are considered signs of acute toxicity and that changes in gross pathology reflect long-term toxicity [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], these parameters were assessed throughout the treatment period. Blood samples were collected from mice administered with i.p. injections of PBS, ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, and the behavior and body weights of these mice were monitored. The results showed that treatment with ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e did not induce significant variations in mouse behavior and body weight compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD), which proved that the acute toxicity of ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was negligible. Similarly, routine blood tests proved that the ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e groups did not cause hemolysis or myelosuppression even after 30 days of treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Further liver and kidney function tests were performed to evaluate the liver or kidney damage induced by ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. ALT and AST were assessed as liver damage indexes, and BUN and CR were assessed as kidney damage indexes [\u003cspan additionalcitationids=\"CR40\" citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. The results demonstrated that both ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e did not damage liver and kidney functions (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB). H\u0026amp;E staining analysis proved that no organ damage was induced by both ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC). All of these results proved that both ZIF-8\u003csup\u003ePt\u003c/sup\u003e and AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e possessed highly biocompatible and potential of clinical transformation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"3. Conclusion","content":"\u003cp\u003eIn this study, a pH-responsive and catalase-like Pt nanoparticle-embedded metal-organic framework (ZIF-8\u003csup\u003ePt\u003c/sup\u003e) loaded with AP (AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e) was developed to treating HCC. Both \u003cem\u003ein vitro\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e experiment results proved that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e had excellent biocompatibility and therapeutic efficacy. Further experiment verified that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e can promote DNA damage and relieving hypoxia. Transcriptomics analysis demonstrated that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e worked through thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, rion disease, PI3K-Akt signaling pathway, purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process and ribonucleoside diphosphate metabolic process, respiratory chain, cytosolic large ribosomal subunit, respiratory chain complex, extracellular mata structin consultent and Oxygen binding. Our study suggested that this nanoplatform AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e offered a promising paradigm for HCC treatment and had clinical application potential.\u003c/p\u003e"},{"header":"4. Materials and Methods","content":"\u003cp\u003e4.1. \u003cem\u003eMaterials\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e1,2-dimethylimidazole (2-MIL), zinc nitrate hexahydrate (Zn (NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e\u0026middot;6H\u003csub\u003e2\u003c/sub\u003eO), chloroplatinic acid hydrate (H\u003csub\u003e2\u003c/sub\u003ePtCl\u003csub\u003e6\u003c/sub\u003e\u0026middot;6H\u003csub\u003e2\u003c/sub\u003eO) and polyvinylpyrrolidone (PVP) were purchased from Aladdin. AP was synthesized by Shanghai Qiangyao Biotechnology Co., Ltd. 4\u0026apos;6-diamidino-2-phenylindole (DAPI) and 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) were purchased from Beyotime Biotechnology. The \u0026gamma;-H2AX antibody, HIF-1\u0026alpha; antibody and secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). All of the reagents used in the experiments were of analytical grade and used directly without treatment.\u003c/p\u003e\n\u003cp\u003e4.2. \u003cem\u003eCharacterizations\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eMalvern Zetasizer Nano ZS90 Apparatus, UV-2550 Ultraviolet-visible Spectrophotometer (Shimadzu, Japan), transmission electron microscopy system (JEOL/JEM-F200), XRD (Bruker/D8 ADVANCE), FTIR (Bruker/VERTEX70), FC500 flow cytometer (Beckman Coulter, USA) were used for material characterization same as our previous studies[26, 42].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e4. 3. \u003cem\u003eSynthesis of\u0026nbsp;\u003c/em\u003e\u003cem\u003eAP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eZinc nitrate hexahydrate (25 mM) and 2-methyl imidazole (200 mM) were first dissolved in methanol. Then, 20 mL of methanol solution was stirred together with Pt nanoparticles. After continuous stirring for 3\u0026ndash;4 h, the sample was washed three times with methanol, and ZIF-8\u003csup\u003ePt\u003c/sup\u003e was obtained. Then redispersed ZIF-8\u003csup\u003ePt\u003c/sup\u003e in ethanol (10 mL) added AP and stirred for 6\u0026ndash;8 h. The mixed solution was then washed with methanol (10000 rpm, 10 min) to remove excess AP then obtained AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e4. 4. \u003cem\u003eIn vitro cytotoxicity assay\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe cytotoxicity against HCC cells was measured by CCK8 assay. The cells were cultured in a 96-well plate and incubated overnight. Next, gradient concentrations of AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e (0, 20, 40, 60, 80 and 100 \u0026mu;g/mL) were added to each well and incubated for 24 h. Five parallel samples were set for each group, then 10 \u0026micro;L of CCK8 was added to each well and incubated for 4 h. The absorbance was quantified by a spectrophotometer at 450 nm.\u003c/p\u003e\n\u003cp\u003e4. 5.\u0026nbsp;\u003cem\u003eR\u003c/em\u003ee\u003cem\u003eactive oxygen species (ROS) detection\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eROS in cells was detected using the ROS probe, DCFH-DA. Briefly, HCC cells were seeded in a 6-well plate and cultured overnight. \u0026nbsp;ZIF-8 and ZIF-8\u003csup\u003ePt\u003c/sup\u003e were added to the cells and incubated in the dark for 2 h. Following incubation, the medium was removed and cells were washed with PBS three times. Following washing, the ROS probe, DCFH-DA, was added to the cells and incubated for a further 30 min, and then the fluorescence of cells generated by DCFH-DA was detected by fluorescence microscopy.\u003c/p\u003e\n\u003cp\u003e4. 6. \u003cem\u003eCellular uptake analysis\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHCC cells were seeded in confocal chambers and cultured overnight. The AP loaded Rhodamine B to track nanoparticles, were added to the chambers and incubated for 2 h. Next, the cells were washed with PBS three times and stained with phalloidin-FITC (30 min) and DAPI (10 min). Finally, the cells were observed by laser confocal microscopy (phalloidin-FITC, excitation: 490 nm; DAPI, excitation: 400 nm, and Rhodamine B, excitation: 555 nm). We further investigated the cellular uptake of nanoparticles by FE-TEM. Briefly, HCC cells were cultured in a cell slide overnight. After incubating with AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e for 2 h, the cell slides were washed with PBS. The cells were fixed with 3% glutaraldehyde overnight and then dehydrated in an ethanol series. Using a mini-gold sputter, cell samples were coated with gold-palladium for visual inspection by TEM.\u003c/p\u003e\n\u003cp\u003e4. 7. \u003cem\u003eFlow cytometry analysis\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHCC cells at a concentration of 1 \u0026times; 10\u003csup\u003e5\u0026nbsp;\u003c/sup\u003ecells were seeded in six-well plates and incubated overnight. Cells were treated with AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, which were loaded with rose red B. After incubation for 3 h and 6h, the cells were washed three times with cold PBS and the treated cells were harvested for flow cytometry analysis.The cells were seeded in 6 well plates at a density of 2\u0026times;10\u003csup\u003e5\u003c/sup\u003e cells and cultured overnight. Apoptosis kit (BD Biosciences, CA) was used for apoptosis analysis and operated according to the instruction.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e4. 8.\u003cem\u003e\u0026nbsp;Immunofluorescence\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHCC cells were fixed with 4% PFA at 37\u0026deg;C for 20 min, treated with 0.02% Triton X-100 for 10 min, and then blocked at room temperature for 1 h. Cells were incubated with primary antibody overnight at 4\u0026deg;C followed by incubation with fluorescent-conjugated secondary antibody for 2 h at room temperature. The nuclei were stained with DAPI for 15 min. The images of the cells were obtained by laser confocal microscopy (\u0026gamma;-H2AX-FITC, excitation: 490 nm, HIF-1\u0026alpha;-CY5, excitation: 650 nm and DAPI, excitation: 400 nm).\u003c/p\u003e\n\u003cp\u003e4. 9. \u003cem\u003eIn vivo therapeutic effect\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eWhen the tumor reached approximately 100 mm\u003csup\u003e3\u003c/sup\u003e, tumor-bearing mice were randomly divided into 4 groups: (I) Control; (II) AP; (III) ZIF-8\u003csup\u003ePt\u003c/sup\u003e, and (IV) AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. After the nanoparticles (50 mg∙kg\u003csup\u003e\u0026minus;1\u003c/sup\u003e) were intravenously injected into the tail veins every 2 d. For treatment, the solid tumor was dissected from one mouse in each group for H\u0026amp;E and TUNEL staining. The solid tumor was also used for HIF-1\u0026alpha; and \u0026gamma;-H2AX IHC staining. The body weights of mice and the tumor volumes were recorded every other day.\u003c/p\u003e\n\u003cp\u003e4. 10. \u003cem\u003eTranscriptomics analysis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eWhen the tumor reached about100 mm\u003csup\u003e3\u003c/sup\u003e, tumor-bearing mice were randomly divided into three groups: (I) Control; (II) ZIF-8\u003csup\u003ePt\u003c/sup\u003e; (III) AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e. Nanoparticles (50 mg/kg) were intravenously injected via tail vein every 2d. At the end of the experiment, tumors were dissected for transcriptomics analysis using an LC-ESI-MS/MS system.\u003c/p\u003e\n\u003cp\u003e4. 16. \u003cem\u003eStatistical analyses\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eOne-way and two-way analyses of variance (ANOVA) and t-tests were used to determine statistical significance (*p\u0026lt;0.05, **p\u0026lt;0.01, ***p\u0026lt;0.001), p-value less than 0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSupporting Information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSupporting Information is available from the Wiley Online Library or from the author.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Declaration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the National Natural Science Foundation of China (No.82403783), the Nature Science Foundation of Fujian Province (No.2024J08316) and National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital \u0026amp; Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen (No.E010322014)\u003cstrong\u003e.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of Interest Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing financial interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics and Consent to Participate declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll animal experiments were approved by the Experimental Animal Management and Ethics Committee of Xiamen University.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eZhenzhen Luo and Dunhuang Wang wrote the main manuscript text; Lie Lin and Rui Zhou prepared figures 1-2. Yuanyuan Su, Zongkai Zhang and Jing Hu prepared figures 3-4; Yaqing Dai and Xiaoyan Huang prepared figure 5; Yufei Zhou and Liuyun Gong reviewed the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCerreto, M., et al., \u003cem\u003eThe New Era of Systemic Treatment for Hepatocellular Carcinoma: From the First Line to the Optimal Sequence.\u003c/em\u003e Curr Oncol, 2023. \u003cstrong\u003e30\u003c/strong\u003e(10): p. 8774-8792.\u003c/li\u003e\n\u003cli\u003eHuang, Z., et al., \u003cem\u003eThe role of long noncoding RNAs in hepatocellular carcinoma.\u003c/em\u003e Mol Cancer, 2020. \u003cstrong\u003e19\u003c/strong\u003e(1): p. 77.\u003c/li\u003e\n\u003cli\u003eCampani, C., J. Zucman-Rossi, and J.C. 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Apoptin (AP)，a coding protein of VP3 gene, stem from the chicken anemia virus (CAV), can be activated in malignant cells selectively and prevents the dividing cancer cells from repairing their DNA lesions, thereby forcing them to undergo apoptosis. Herein, a three-step intelligent biodegradable drug delivery nanoplatform was designed. First, a hollow ZIF-8 was synthesized, embedded with platinum nanoparticle to form ZIF-8\u003csup\u003ePt\u003c/sup\u003e, and then loaded with AP, and lastly formed AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e, which possess pH-responsive drug release and cancer-targeted ability. As expected, both in vitro and in vivo experiment demonstrated that AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e performed treatment effects in hepatocarcinoma through relieving tumor-hypoxic microenvironment, inhibiting cell proliferation, and promoting cell apoptosis. Further ranscriptomic analysis showed that the specific mechanism of the AP@ZIF-8\u003csup\u003ePt\u003c/sup\u003e was thermogenesis, signaling pathways regulating pluripotency of stem cells, ribosome, prion disease and PI3K-Akt signaling pathway.\u0026nbsp;\u0026nbsp; This work highlights a new strategy for liver cancer treatment and provides a reference for treating malignant tumors.\u003c/p\u003e","manuscriptTitle":"A super-assembled synergistically nanoplatform AP@ZIF-8 Pt for hepatocarcinoma therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-02 08:08:51","doi":"10.21203/rs.3.rs-5708497/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0928d880-943a-4c88-a08b-7f9ce9505010","owner":[],"postedDate":"January 2nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-01-02T17:08:41+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-02 08:08:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5708497","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5708497","identity":"rs-5708497","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}