DNA Nanoflower Enables Controlled Co-Delivery of Antisense Oligodeoxynucleotide and Doxorubicin for Anti-breast Cancer Treatment

DNA Nanoflower Enables Controlled Co-Delivery of Antisense Oligodeoxynucleotide and Doxorubicin for Anti-breast Cancer Treatment | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL Micro & Nano Letters This is a preprint and has not been peer reviewed. Data may be preliminary. 21 February 2025 V1 Latest version Share on DNA Nanoflower Enables Controlled Co-Delivery of Antisense Oligodeoxynucleotide and Doxorubicin for Anti-breast Cancer Treatment Authors : Xiuping Shen 0009-0005-9985-0260 [email protected] , Aiyong Zhu , and Yafeng Xu Authors Info & Affiliations https://doi.org/10.22541/au.174017526.63124569/v1 Published Micro & Nano Letters Version of record Peer review timeline 385 views 190 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Doxorubicin (DOX), an anthracycline antibiotic, is widely used to treat a range of solid tumors and hematological malignancies. However, its clinical application in breast cancer is hindered by toxic side effects and the development of multidrug resistance (MDR). Enhancing the selective targeting of DOX and overcoming MDR are critical to improving treatment efficacy. Here, we present a DNA nanoflower (DNF)-based delivery system, designed via rolling circle amplification (RCA) and multi-primer amplification (MCA), which co-delivers antisense oligonucleotides (ASO) and DOX to human breast cancer cells (MCF-7). This system, named DNF-ASO@DOX, effectively promotes gene silencing, enhances drug accumulation, and significantly inhibits cell proliferation. Furthermore, in vivo studies using mouse models of breast cancer demonstrated potent therapeutic effects, highlighting DNF-ASO@DOX as a promising strategy for enhanced anti-tumor therapy. DNA Nanoflower Enables Controlled Co-Delivery of Antisense Oligodeoxynucleotide and Doxorubicin for Anti-breast Cancer Treatment Xiuping Shen a,* , Aiyong Zhu b , Yafeng Xu c a School of Pharmacy, Kunming Medical University, Kunming 650500, China e-mail: [email protected] b Shanghai University of Medicine & Health Sciences, Shanghai, 201318, China c Sinopharm Nursing Care Industrial (Shanghai) Co., LTD, Shanghai, 200333, China Correspondence: Xiuping Shen; e-mail: [email protected] ; School of Pharmacy, Kunming Medical University Abstract: Doxorubicin (DOX), an anthracycline antibiotic, is widely used to treat a range of solid tumors and hematological malignancies. However, its clinical application in breast cancer is hindered by toxic side effects and the development of multidrug resistance (MDR). Enhancing the selective targeting of DOX and overcoming MDR are critical to improving treatment efficacy. Here, we present a DNA nanoflower (DNF)-based delivery system, designed via rolling circle amplification (RCA) and multi-primer amplification (MCA), which co-delivers antisense oligonucleotides (ASO) and DOX to human breast cancer cells (MCF-7). This system, named DNF-ASO@DOX, effectively promotes gene silencing, enhances drug accumulation, and significantly inhibits cell proliferation. Furthermore, in vivo studies using mouse models of breast cancer demonstrated potent therapeutic effects, highlighting DNF-ASO@DOX as a promising strategy for enhanced anti-tumor therapy. Keywords: DNA nanoflower; Rolling circle amplification; Multi-primer amplification; Antisense oligonucleotide; Breast cancer treatment 1.Introduction Cancer remains a leading cause of global mortality, significantly hindering life expectancy improvements worldwide [1]. By 2020, female breast cancer surpassed lung cancer as the most common cancer globally, accounting for 11.7% of all cancer cases, Breast cancer (BC) is the most prevalent malignancy among women, and its pathogenesis is still not fully understood [2,3]. Chemotherapy remains the standard treatment for malignant tumors [4,5], with doxorubicin (DOX) being one of the most effective anti-cancer agents. However, DOX faces limitations such as short half-life, cardiotoxicity, multidrug resistance (MDR), and poor prognosis [6-9]. Thus, there is a critical need for drug delivery systems that enhance targeting, reduce toxicity, and overcome MDR. Polo-like kinase 1 (PLK1), a key regulator of mitosis and DNA replication, is frequently overexpressed in various malignant tumors, correlating with advanced stages and poor prognosis [10-14]. Targeting PLK1 has emerged as a promising strategy for tumor therapy, as its inhibition can disrupt the cell cycle and enhance sensitivity to treatments like radiotherapy [15-17]. Gene therapy, particularly using small nucleic acid drugs, has gained attention for treating cancers, metabolic disorders, and genetic diseases [18-20]. Antisense oligonucleotides (ASOs), which bind RNA target sequences through Watson-Crick base pairing, offer multiple therapeutic possibilities, including RNA degradation, splicing alterations, and gene editing [21]. However, efficient delivery to non-hepatic tissues remains a significant challenge [21]. DNA, a versatile molecular building block, has been used to design multifunctional nanostructures such as DNA origami, dendrimers, and nanocages [22-27]. These DNA-based nanostructures are biocompatible, programmable, and structurally predictable, making them ideal candidates for applications in diagnostics and drug delivery [28]. In cancer therapy, DNA nanocarriers offer substantial advantages, including high nucleic acid drug loading capacity [29-31]. DNA nanoflowers (DNFs), DNA/inorganic hybrid nanoparticles with flower-like morphology, possess unique properties, such as a porous structure and easily modifiable DNA strands, enabling them to serve as efficient carriers for combined gene silencing and chemotherapy [36-39]. Here, we report a multifunctional DNA nanoflower (DNF) framework capable of co-delivering antisense oligonucleotides (ASO) and doxorubicin (DOX), termed DNF-ASO@DOX, for the synergistic treatment of breast cancer. DNF was developed on the basis of a rolling circle amplification (RCA) reaction and a multiplexed enzyme amplification (MCA) reaction with the addition of Phi29 DNA polymerase developed on the basis of the rolling circle amplification (RCA) reaction and the multimeric enzyme amplification (MCA) reaction with the addition of Phi29 DNA polymerase. This DNF-ASO@DOX combines AS1411 aptamer, ASO and DOX. AS1411 aptamer has been specifically designed to increase cellular uptake and improve tumor targeting. DNF is responsive to specific endonuclease 1 (FEN1), which is overexpressed in cancer cells [40]. ASO can be cleaved and released from DNF via FEN1 for gene silencing. that is used for gene silencing. At the same time, DNF can release DOX in tumor tissues, increasing local drug concentration and therapeutic efficiency, reducing side effects and MDR of tumor cells. This synergistic therapy combining gene silencing with chemotherapy greatly improves anti-tumor efficacy in vitro and in vivo. 2. Results and Discussion 2.1 Anti-cancer mechanism of DNF-ASO@DOX DNA nanoflowers (DNFs) were synthesized using a rolling circle amplification (RCA) reaction with Phi29 DNA polymerase, followed by a multi-primer amplification (MCA) reaction to construct the DNF-ASO nanocomplex. As illustrated in Figure 1, DNF served as the core framework to facilitate the loading of doxorubicin (DOX). Tumor-targeting long-stranded DNA was synthesized by encoding complementary AS1411 aptamer sequences and antisense oligonucleotide (ASO) sequences within a loop template. Circular DNA was formed through cyclization with T4 DNA ligase, followed by the Phi29 polymerase-catalyzed RCA reaction to yield the DNF. The MCA reaction was subsequently used to prepare DNF-ASO, which exhibited a porous structure that enhanced its drug-loading capacity. After preparing DNF-ASO, DOX was loaded into the nanocomplex, resulting in the formation of DNF-ASO@DOX. This system combines two active components—ASO and DOX—ensuring structural adaptability for efficient drug delivery and release. Upon cellular uptake, the DNF-ASO@DOX complex facilitates the controlled release of both ASO and DOX at the tumor site. ASO contributes to gene silencing, while DOX exerts its chemotherapeutic effects, inducing apoptosis in cancer cells. This dual mechanism of action enables synergistic treatment, where gene silencing and targeted drug delivery work in concert to inhibit tumor cell proliferation Figure 1. DNF-ASO@DOX was used for controlled co-administration of ASO and DOX for synergistic treatment of breast cancer. A) Preparation of DNF efficiently loaded with ASO and DOX. B) Cellular uptake of the nanocomplex and controlled release of ASO and DOX. Synergistic treatment was achieved by gene silencing and chemotherapy. 2.2 Synthesis and Characterization of DNF DNF was synthesized by cyclizing the DNA template using T4 DNA ligase, followed by rolling circle amplification (RCA) with Phi29 DNA polymerase. Polyacrylamide gel electrophoresis (PAGE) analysis revealed that the molecular weight of the cyclic DNA was greater than that of the DNA template and primers, and that the RCA product exhibited a higher molecular weight than the cyclic DNA (Fig. 2A). These results confirmed successful synthesis of cyclic DNA and DNF. Further characterization of DNF was performed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). SEM and TEM images (Fig. 2B and 2C) showed that the DNF exhibited a porous, flower-like structure composed of intersecting nanosheets approximately 500 ± 50 nm in size. The unique porous structure of DNF contributed to its high drug-loading efficiency and enhanced its ability to target tumor cells and improve cellular uptake. Figure 2. Synthesis and characterization of DNF. A) PAGE analysis for the synthesis of DNF. B) SEM images of DNF. C) TEM images of DNF. 2.3 Synthesis and Characterization of DNF-ASO Following the successful synthesis of DNF, multi-primer amplification (MCA) was used to prepare DNF-ASO, utilizing primers 2 and 3. The impact of dNTP concentration and reaction time on the morphology and size of DNF-ASO was systematically evaluated. Under optimal conditions, DNF-ASO exhibited a size of approximately 400 ± 50 nm, which facilitated its accumulation at the tumor site and improved cellular uptake due to its nanoscale pore size. PAGE electrophoresis confirmed that the molecular weight of the cyclic DNA was greater than that of the DNA template and primers, and that the molecular weight of the MCA product exceeded that of the cyclic DNA (Fig. 3). SEM and TEM analysis revealed that DNF-ASO displayed a porous, flower-like structure composed of intersecting nanosheets, with a size of approximately 400 ± 50 nm (Fig. 3B-C). These findings suggest that the DNF-ASO nanocomplex is well-suited for efficient drug delivery and enhanced tumor targeting. Figure 3. Synthesis and characterization of DNF-ASO. A) PAGE analysis for the synthesis of DNF-ASO. B) SEM images of DNF-ASO. C) TEM images of DNF-ASO. 2.4 Drug Loading The unique porous flower-like structure of DNF-ASO enabled efficient drug loading, improved cellular uptake, and enhanced anti-tumor activity. For drug loading, DNF-ASO (50000 ng) was dispersed in 100 μL ddH 2 O, and then DOX (5 mg mL -1 , 1 μL) was added. The mixture was incubated at 4 °C for 3 hours to ensure effective encapsulation of DOX in the mesopores, resulting in the formation of the final DNF-ASO@DOX complex. The DNF-ASO@DOX complex was collected via centrifugation, extensively washed with ddH 2 O to remove any unloaded DOX, and the fluorescence of the supernatant was measured at an excitation wavelength of 488 nm and an emission wavelength of 550 nm. Drug-loading content (DLC) and drug-loading efficiency (DLE) were calculated using equations (1) and (2): DLC = (weight of drug loaded in DNF/weight of DNF) × 100% (1); DLE = (weight of drug loaded in DNF/weight of feed drug) × 100% (2). 2.5 Cellular Uptake and Intracellular Distribution of DNF-ASO To assess the gene silencing and chemotherapeutic effects of DNF-ASO, MCF-7 cells, a human breast cancer cell line, were treated with Cy5-labeled DNF-ASO. Cellular distribution was analyzed using laser scanning confocal microscopy (LSCM) (Fig. 4A). The cells were incubated with DNF-ASO for varying time points, and DAPI (blue) was used to visualize the nuclei. LSCM images revealed that the fluorescence intensity of Cy5 gradually increased over time, reaching a stable level within 12 hours and peaking at 24 hours, confirming the effective uptake of DNF-ASO by MCF-7 cells. Co-localization of the red (Cy5) and blue (DAPI) signals was observed in the LSCM images, indicating that DNF-ASO was entering the nucleus. Initially, the red signal overlapped with the green signal within the first 4 hours. After 6 hours, the red signal began to separate from the green, and by 24 hours, the red and blue signals were almost completely separated, suggesting that DNF-ASO had been fully internalized by the MCF-7 cells by this time (Fig. 4B). Figure 4. Cellular uptake and intracellular distribution of DNF-ASO. A) LSCM images of nuclei co-localization. Blue, DAPA -stained nuclei; red, Cy5-labeled DNF-ASO. B) Statistical analysis of DNF-ASO uptake efficiency. Data represented as mean ± SD, n=3, ***P<0.001. To further verify whether DNF-ASO could be targeted and absorbed by MCF-7 cells, we performed further validation using flow cytometry (Figure 5). The results of flow cytometry analysis showed that the uptake rate gradually increased with the prolongation of uptake time (Fig. 5A), reaching 34.72 ± 1.57% at 6 h, 72.35 ± 0.48% at 12 h, and 93.35 ± 0.40% at 24 h, suggesting that DNF-ASO essentially enters into the cell within 24 h. Meanwhile, similar results (Figure 5B) were further validated by the number of cells containing Cy5-labelled DNF-ASO. Figure 5. The flow cytometry analysis for cellular uptake of the DNF-ASO in MCF-7 cells. A) The uptake rate analysis of MCF-7 cells treated with DNF-ASO using flow cytometry with Annexin V-FITC/PI assay. B) The number of cells containing Cy5-labeled DNF-ASO. Data represented as mean ± SD, n=3, *P<0.05, ***P<0.001. 2.6 Synergistic Therapeutic Effect of DNF-ASO@DOX In Vitro To evaluate the therapeutic effects, MCF-7 cells were treated with three nanocomplexes: DNF (used as a control, where the ASO was replaced with a scrambled sequence to assess the biocompatibility of DNA nanoflowers), DNF-ASO (designed for PLK1 gene silencing), and DNF-ASO@DOX (a combination of gene silencing and chemotherapy, with DOX loaded onto DNF containing ASO sequences). The DNF-ASO@DOX complex was designed to control the release of both ASO and DOX for a synergistic therapeutic effect. The enzyme-specific release mechanism was triggered by the overexpressed nuclease endonuclease 1 (FEN1) in cancer cells, which cleaves the ASO from DNF, silencing the PLK1 gene and enhancing DOX delivery to the tumor site. PLK1 is overexpressed in breast cancer and is associated with poor prognosis. Inhibition of PLK1 expression suppresses tumor cell proliferation and increases the sensitivity of tumor cells to chemotherapy. MCF-7 cells were treated with various concentrations of DNF, DNF-ASO, DNF-ASO@DOX, and DOX for 48 hours, and cell viability was measured by CCK-8 assay. The results showed no significant cytotoxicity in the DNF-treated group, indicating good biocompatibility (Fig. 6A). However, the viability of cells treated with DNF-ASO, DNF-ASO@DOX, and DOX decreased in a dose-dependent manner, suggesting potent therapeutic effects of these formulations. The IC 50 value for DNF-ASO@DOX was determined to be 9.296 ng/μL (Fig. 6B). When treated at 20 ng/μL, the lowest survival rate was observed in the DNF-ASO@DOX-treated group (46.89 ± 0.82%), compared to the DNF-ASO (72.36 ± 1.21%) and DOX (60.91 ± 1.29%) groups, confirming the synergistic therapeutic effect of ASO and DOX (Fig. 6C). Additionally, flow cytometry analysis of apoptosis, assessed by Annexin-V-FITC/PI staining, revealed a higher apoptosis rate in the DNF-ASO@DOX-treated group (60.60 ± 0.95%) compared to the DNF-ASO-treated group (13.57 ± 0.36%) and the DOX-treated group (45.23 ± 2.46%) (Fig. 6D). These findings underscore the enhanced efficacy of the combined gene silencing and chemotherapy approach. Figure 6. Gene silencing and chemotherapeutic effects in vitro. A) Survival rate of MCF-7 cells treated with different drug formulations. B) IC 50 value of DNF-ASO@DOX. C) Viability of MCF-7 cells treated with different drug formulations at a concentration of 20 ng μL -1 . D) Apoptosis analysis of MCF-7 cells treated with different drug formulations by using flow cytometry and Annexin V-FITC/PI assays. E) MCF-7 cells treated with different drug formulations were analyzed for apoptosis using flow cytometry and Annexin V-FITC/PI assay. Data were expressed as mean ± SD, n=3, **P<0.01, ***P<0.001. Cell cycle assay and cell migration assay were used to further evaluate the therapeutic effects of DNF-ASO@DOX in vitro. Cell cycle analysis by flow cytometry showed that cells were arrested at G2/M phase after treatment with DNF-ASO, DNF-ASO@DOX and DOX, which indicated that DNF-ASO and DNF-ASO@DOX promoted cell differentiation and induced DNA damage during the differentiation phase of the cell cycle (Figure 7A). Meanwhile, compared with the PBS-treated group, the DNF-treated group did not affect the cell cycle, and the DNF-ASO, DNF-ASO@DOX and DOX-treated groups all significantly interfered with the cell cycle, with DNF-ASO@DOX interfering with the cell cycle most dramatically, showing a synergistic inhibitory effect on cell proliferation. The results of cell migration showed that the number of cells entering the lower lumen in the DNF-treated group (78±2.34) was comparable to that in the PBS-treated group (75±08.20), indicating that DNF did not weaken the migratory and invasive ability of MCF-7 cells (Figure 7B).Compared with the PBS and DNF-treated groups, the number of cells entering the lower lumen was significantly reduced in the DNF-ASO (60±1.03), DNF-ASO@DOX (14±0.35) and DOX (38±2.25) treated groups, and was significantly less in the DNF-ASO@DOX-treated group compared with the other two groups, suggesting that DNF-ASO@DOX synergistically attenuated MCF-7 cells’ migration and invasion ability. The effect of gene silencing was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunoblotting (WB). The relative expression levels of PLK1 mRNA showed negligible differences between the PBS and DNF-treated groups. The expression of PLK1 mRNA showed a decreasing trend in the DNF-ASO, DNF-ASO@DOX and DOX-treated groups (respectively 30.33±0.09%, 12.53±0.07% and 35.63±0.02%, respectively), confirming the high efficiency of the synergistic treatment (Figure 7C). Similar results were found by WB analysis (Figure 7D). In MCF-7 cells, the expression level of PLK1 protein showed the same trend among the three treatment groups, and the relative expression level of PLK1 protein was the lowest in the DNF-ASO@DOX-treated group (relative to the DNF-ASO and DOX-treated groups). These results demonstrate that the synergistic effect of gene silencing and chemotherapy can most effectively downregulate PLK1. Figure 7. Gene silencing and Chemotherapeutic effects in vitro. A) Cell cycle analysis of MCF-7 cells treated with different drug formulations using flow cytometry and Annexin V-FITC/PI assay. B) Crystal violet staining to analyse the cell migration rate. C) qRT-PCR to analyse the relative expression level of PLK1 mRNA. D) WB analysis of PLK1 protein expression. Data were expressed as mean ± SD, n = 3, *P < 0.05, **P < 0.01, ***P < 0.001 . 2.7 Synergistic Therapeutic Effect of DNF-ASO@DOX In Vivo The therapeutic efficacy of DNF-ASO@DOX in vivo was assessed using BALB/c nude mice with MCF-7 xenografts. Mice were treated with saline, DNF, DNF-ASO, DNF-ASO@DOX, or DOX, with local administration of 2 mg/kg doses on days 0, 3, and 6 when tumors reached 100 mm³. Mice were monitored every two days for 14 days (Fig. 8A). Body weight measurements indicated minimal differences between the saline and DNF-treated groups, confirming the biocompatibility of DNF (Fig. S1, Supporting Information). Tumor volume was recorded and tumor growth curves were plotted (Fig. 8B), showing negligible differences between the saline and DNF groups (Fig. 8D and Fig. S2). Tumor volume decreased slightly in the DNF-ASO and DOX-treated groups, with the most significant suppression observed in the DNF-ASO@DOX-treated group, which exhibited minimal tumor growth. Tumors from the DNF-ASO@DOX-treated group had the smallest volumes and demonstrated the most effective therapeutic response, outperforming both DOX and DNF-ASO alone. These results were further confirmed by tumor weight measurements (Fig. 8F). The tumor inhibition rate in the DNF-ASO@DOX group was 64.94 ± 0.08%, significantly higher than the rates for DNF-ASO (32.92 ± 0.13%) and DOX (48.73 ± 0.13%) treatments, indicating the synergistic effect of gene silencing and chemotherapy. After 14 days of treatment, the biocompatibility of DNF was evaluated. Serum biochemical analysis and histological examination of major organs via hematoxylin and eosin (H&E) staining showed minimal differences between the DNF, DNF-ASO, and DNF-ASO@DOX groups, with no obvious tissue necrosis, confirming the good biocompatibility of the treatments (Fig. S3, S4). Immunohistochemical (IHC) analysis revealed significant downregulation of PLK1 expression in tumors from DNF-ASO and DOX groups, with the most pronounced reduction observed in the DNF-ASO@DOX-treated group (Fig. 8G). Western blotting (WB) assays also showed the greatest downregulation of PLK1 protein expression in the DNF-ASO@DOX-treated group (Fig. 8C), further supporting the synergistic effect of gene silencing and chemotherapy. Tissue apoptosis was assessed using TdT-mediated dUTP nick end labeling (TUNEL) and H&E staining (Fig. 8H, 8I). The saline and DNF-treated groups exhibited normal tissue morphology, while the DNF-ASO-treated group showed signs of abnormal cell morphology and extensive tissue necrosis. The DOX-treated group displayed moderate apoptosis, but the DNF-ASO@DOX-treated group exhibited the most extensive tissue necrosis and apoptotic cell death, indicating the most effective anti-tumor effect. These results collectively demonstrate that DNF-ASO@DOX provides a potent synergistic effect of gene silencing and chemotherapy, leading to enhanced anti-tumor efficacy in vivo. Figure 8. Gene silencing and chemotherapeutic effects in vivo. A) Schematic of the treatment process in tumor-bearing mice. B) Tumor growth curves of each mouse in different treatment groups. C) WB analysis of PLK1 protein expression in tumor xenografts. D) Average growth curves of the tumors in different treatment groups. E) Images of resected tumors in mice in different treatment groups. F) Average tumor weights of the tumor groups. G) IHC staining images of PLK1 expression in tumor xenografts. H) TUNEL staining images of tumor xenografts. I) H&E staining images of tumor xenografts. Data are expressed as mean ± SD, n = 5, *P < 0.05, **P < 0.01, ***P < 0.001. 3. Conclusion This study presents the DNF-ASO@DOX delivery system as a novel approach for synergistically treating breast cancer by co-administering antisense oligonucleotides (ASO) and doxorubicin (DOX). The system utilizes nuclease-triggered release of ASO to silence PLK1 mRNA and facilitates targeted DOX delivery to tumor sites. Both in vitro and in vivo results demonstrated significant anti-tumor efficacy, with minimal toxicity to normal tissues, confirming the biocompatibility of DNF. The DNF-based system’s versatility in targeted cell delivery and controlled drug release presents a promising new strategy for enhancing cancer therapy. Conflict of Interest The authors declare no conflict of interest. Author contributions statement Xiuping Shen: writing - original draft and data curation, Aiyong Zhu: revision, Yafeng Xu: conceptualization. All authors have read and agreed to the published version of the manuscript. All authors have read and agreed to the published version of the manuscript. Funding The authors have no funding to disclose at this time. Institutional Review Board Statement All animal experiments were approved by the ethics committee of Kunming Medical University in compliance with the Animal Management Rules of the Ministry of Health of the People’s Republic of China (approval number: KMMUD20230005). References [1] F. Bray, M. Laversanne, E. Weiderpass, I. Soerjomataram, The ever-increasing importance of cancer as a leading cause of premature death worldwide, Cancer 127 (2021) 3029–3030. https://doi.org/10.1002/cncr.33587.[2] H. Sung, J. Ferlay, R.L. Siegel, M. Laversanne, I. Soerjomataram, A. Jemal, F. Bray, Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries, CA A Cancer J Clinicians 71 (2021) 209–249. https://doi.org/10.3322/caac.21660.[3] E. Alphandéry, Perspectives of Breast Cancer Thermotherapies, Journal of Cancer 5 (2014).[4] E.S. McDonald, A.S. Clark, J. Tchou, P. Zhang, G.M. Freedman, Clinical Diagnosis and Management of Breast Cancer, J Nucl Med 57 (2016) 9S-16S. https://doi.org/10.2967/jnumed.115.157834.[5] M.E.R. O’Brien, A. Borthwick, A. Rigg, A. Leary, L. Assersohn, K. Last, S. Tan, S. Milan, D. Tait, I.E. Smith, Mortality within 30 days of chemotherapy: a clinical governance benchmarking issue for oncology patients, Br J Cancer 95 (2006) 1632–1636. https://doi.org/10.1038/sj.bjc.6603498.[6] R. Luo, Distinct biodistribution of doxorubicin and the altered dispositions mediated by different liposomal formulations, International Journal of Pharmaceutics (2017).[7] C.F. Thorn, C. Oshiro, S. Marsh, T. Hernandez-Boussard, H. McLeod, T.E. Klein, R.B. Altman, Doxorubicin pathways: pharmacodynamics and adverse effects, Pharmacogenet Genomics 21 (2011) 440–446. https://doi.org/10.1097/FPC.0b013e32833ffb56.[8] Y.-W. Eom, M.A. Kim, S.S. Park, M.J. Goo, H.J. Kwon, S. Sohn, W.-H. Kim, G. Yoon, K.S. Choi, Two distinct modes of cell death induced by doxorubicin: apoptosis and cell death through mitotic catastrophe accompanied by senescence-like phenotype, Oncogene 24 (2005) 4765–4777. https://doi.org/10.1038/sj.onc.1208627.[9] H.J. Broxterman, K.J. Gotink, H.M.W. Verheul, Understanding the causes of multidrug resistance in cancer: a comparison of doxorubicin and sunitinib, Drug Resist Updat 12 (2009) 114–126. https://doi.org/10.1016/j.drup.2009.07.001.[10] J. Zhang, L. Zhang, J. Wang, L. Ouyang, Y. Wang, Polo-like Kinase 1 Inhibitors in Human Cancer Therapy: Development and Therapeutic Potential, J Med Chem 65 (2022) 10133–10160. https://doi.org/10.1021/acs.jmedchem.2c00614.[11] J. Guan, P. Liu, A. Wang, B. Wang, Long non‑coding RNA ZEB2‑AS1 affects cell proliferation and apoptosis via the miR‑122‑5p/PLK1 axis in acute myeloid leukemia, Int J Mol Med 46 (2020) 1490–1500. https://doi.org/10.3892/ijmm.2020.4683.[12] Z. Li, Y. Zhang, Y. Zhou, F. Wang, C. Yin, L. Ding, S. Zhang, Tanshinone IIA suppresses the progression of lung adenocarcinoma through regulating CCNA2-CDK2 complex and AURKA/PLK1 pathway, Sci Rep 11 (2021) 23681. https://doi.org/10.1038/s41598-021-03166-2.[13] S. Chakraborty, C.Y. Park, Pathogenic Mechanisms in Acute Myeloid Leukemia, Curr Treat Options Oncol 23 (2022) 1522–1534. https://doi.org/10.1007/s11864-022-01021-8.[14] M.E. Salama, D.A. Khairy, Polo-Like Kinase 1(PLK1) Immunohistochemical Expression in Triple Negative Breast Carcinoma: A Probable Therapeutic Target, Asian Pac J Cancer Prev 22 (2021) 3921–3925. https://doi.org/10.31557/APJCP.2021.22.12.3921.[15] L. Gheghiani, L. Wang, Y. Zhang, X.T.R. Moore, J. Zhang, S.C. Smith, Y. Tian, L. Wang, K. Turner, C.K. Jackson-Cook, N.D. Mukhopadhyay, Z. Fu, PLK1 Induces Chromosomal Instability and Overrides Cell-Cycle Checkpoints to Drive Tumorigenesis, Cancer Res 81 (2021) 1293–1307. https://doi.org/10.1158/0008-5472.CAN-20-1377.[16] L. Cirillo, Y. Thomas, L. Pintard, M. Gotta, BORA-dependent PLK1 regulation: A new weapon for cancer therapy?, Mol Cell Oncol 3 (2016) e1199265. https://doi.org/10.1080/23723556.2016.1199265.[17] R.E.A. Gutteridge, M.A. Ndiaye, X. Liu, N. Ahmad, Plk1 Inhibitors in Cancer Therapy: From Laboratory to Clinics, Mol Cancer Ther 15 (2016) 1427–1435. https://doi.org/10.1158/1535-7163.MCT-15-0897.[18] Y. Yamada, Nucleic Acid Drugs—Current Status, Issues, and Expectations for Exosomes, Cancers 13 (2021) 5002. https://doi.org/10.3390/cancers13195002.[19] J.A. Roth, R.J. Cristiano, Gene Therapy for Cancer: What Have We Done and Where Are We Going?, JNCI Journal of the National Cancer Institute 89 (1997) 21–39. https://doi.org/10.1093/jnci/89.1.21.[20] X. Tan, F. Jia, P. Wang, K. Zhang, Nucleic acid-based drug delivery strategies, Journal of Controlled Release 323 (2020) 240–252. https://doi.org/10.1016/j.jconrel.2020.03.040.[21] T.C. Roberts, R. Langer, M.J.A. Wood, Advances in oligonucleotide drug delivery, Nat Rev Drug Discov 19 (2020) 673–694. https://doi.org/10.1038/s41573-020-0075-7.[22] J. Li, A.A. Green, H. Yan, C. Fan, Engineering nucleic acid structures for programmable molecular circuitry and intracellular biocomputation, Nat Chem 9 (2017) 1056–1067. https://doi.org/10.1038/nchem.2852.[23] G. Zhang, S.P. Surwade, F. Zhou, H. Liu, DNA nanostructure meets nanofabrication, Chem Soc Rev 42 (2013) 2488–2496. https://doi.org/10.1039/c2cs35302d.[24] F. Li, J. Tang, J. Geng, D. Luo, D. Yang, Polymeric DNA hydrogel: Design, synthesis and applications, Progress in Polymer Science 98 (2019) 101163. https://doi.org/10.1016/j.progpolymsci.2019.101163.[25] S. Nummelin, J. Kommeri, M.A. Kostiainen, V. Linko, Evolution of Structural DNA Nanotechnology, Adv Mater 30 (2018) e1703721. https://doi.org/10.1002/adma.201703721.[26] D. Yang, M.R. Hartman, T.L. Derrien, S. Hamada, D. An, K.G. Yancey, R. Cheng, M. Ma, D. Luo, DNA materials: bridging nanotechnology and biotechnology, Acc Chem Res 47 (2014) 1902–1911. https://doi.org/10.1021/ar5001082.[27] Y. Dong, C. Yao, Y. Zhu, L. Yang, D. Luo, D. Yang, DNA Functional Materials Assembled from Branched DNA: Design, Synthesis, and Applications, Chem Rev 120 (2020) 9420–9481. https://doi.org/10.1021/acs.chemrev.0c00294.[28] Q. Hu, H. Li, L. Wang, H. Gu, C. Fan, DNA Nanotechnology-Enabled Drug Delivery Systems, Chem Rev 119 (2019) 6459–6506. https://doi.org/10.1021/acs.chemrev.7b00663.[29] W. Zhou, R. Saran, J. Liu, Metal Sensing by DNA, Chem Rev 117 (2017) 8272–8325. https://doi.org/10.1021/acs.chemrev.7b00063.[30] J. Li, C. Zheng, S. Cansiz, C. Wu, J. Xu, C. Cui, Y. Liu, W. Hou, Y. Wang, L. Zhang, I. -ting Teng, H.-H. Yang, W. Tan, Self-assembly of DNA nanohydrogels with controllable size and stimuli-responsive property for targeted gene regulation therapy, J Am Chem Soc 137 (2015) 1412–1415. https://doi.org/10.1021/ja512293f.[31] M. Kwak, A. Herrmann, Nucleic acid amphiphiles: synthesis and self-assembled nanostructures, Chem Soc Rev 40 (2011) 5745–5755. https://doi.org/10.1039/c1cs15138j.[32] Supramolecular Self‐Assembled DNA Nanosystem for Synergistic Chemical and Gene Regulations on Cancer Cells - Li - 2021 - Angewandte Chemie International Edition - Wiley Online Library, (n.d.). https://onlinelibrary.wiley.com/doi/10.1002/anie.202111900 (accessed November 5, 2024).[33] J. Liu, T. Wu, X. Lu, X. Wu, S. Liu, S. Zhao, X. Xu, B. Ding, A Self-Assembled Platform Based on Branched DNA for sgRNA/Cas9/Antisense Delivery, J. Am. Chem. Soc. 141 (2019) 19032–19037. https://doi.org/10.1021/jacs.9b09043.[34] A DNA Nanocomplex Containing Cascade DNAzymes and Promoter‐Like Zn‐Mn‐Ferrite for Combined Gene/Chemo‐dynamic Therapy - Yao - 2022 - Angewandte Chemie International Edition-Wiley Online Library, (n.d.). https://onlinelibrary.wiley.com/doi/10.1002/anie.202113619 (accessed November 4, 2024).[35] J. Wang, H. Wang, H. Wang, S. He, R. Li, Z. Deng, X. Liu, F. Wang, Nonviolent Self-Catabolic DNAzyme Nanosponges for Smart Anticancer Drug Delivery, ACS Nano 13 (2019) 5852–5863. https://doi.org/10.1021/acsnano.9b01589.[36] J.B. Lee, S. Peng, D. Yang, Y.H. Roh, H. Funabashi, N. Park, E.J. Rice, L. Chen, R. Long, M. Wu, D. Luo, A mechanical metamaterial made from a DNA hydrogel, Nature Nanotech 7 (2012) 816–820. https://doi.org/10.1038/nnano.2012.211.[37] P. Shende, P. Kasture, R.S. Gaud, Nanoflowers: the future trend of nanotechnology for multi-applications, Artif Cells Nanomed Biotechnol 46 (2018) 413–422. https://doi.org/10.1080/21691401.2018.1428812.[38] N. Zhao, L. Yan, X. Zhao, X. Chen, A. Li, D. Zheng, X. Zhou, X. Dai, F.-J. Xu, Versatile Types of Organic/Inorganic Nanohybrids: From Strategic Design to Biomedical Applications, Chem Rev 119 (2019) 1666–1762. https://doi.org/10.1021/acs.chemrev.8b00401.[39] Y. Liu, X. Ji, Z. He, Organic-inorganic nanoflowers: from design strategy to biomedical applications, Nanoscale 11 (2019) 17179–17194. https://doi.org/10.1039/c9nr05446d.[40] H. Zhang, S. Ba, D. Mahajan, J.Y. Lee, R. Ye, F. Shao, L. Lu, T. Li, Versatile Types of DNA-Based Nanobiosensors for Specific Detection of Cancer Biomarker FEN1 in Living Cells and Cell-Free Systems, Nano Lett 18 (2018) 7383–7388. https://doi.org/10.1021/acs.nanolett.8b03724. Information & Authors Information Version history V1 Version 1 21 February 2025 Peer review timeline Published Micro & Nano Letters Version of Record 1 Jul 2025 Published Copyright This work is licensed under a Non Exclusive No Reuse License. Collection Micro & Nano Letters Keywords biomedical materials cancer nanomedicine Authors Affiliations Xiuping Shen 0009-0005-9985-0260 [email protected] Kunming Medical University - Chenggong Campus View all articles by this author Aiyong Zhu Shanghai University of Medicine and Health Sciences View all articles by this author Yafeng Xu Sinopharm Group Co Ltd View all articles by this author Metrics & Citations Metrics Article Usage 385 views 190 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Xiuping Shen, Aiyong Zhu, Yafeng Xu. DNA Nanoflower Enables Controlled Co-Delivery of Antisense Oligodeoxynucleotide and Doxorubicin for Anti-breast Cancer Treatment. Authorea . 21 February 2025. DOI: https://doi.org/10.22541/au.174017526.63124569/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . Format Please select one from the list RIS (ProCite, Reference Manager) EndNote BibTex Medlars RefWorks Direct import Tips for downloading citations document.getElementById('citMgrHelpLink').addEventListener('click', function() { popupHelp(this.href); return false; }); $(".js__slcInclude").on("change", function(e){ if ($(this).val() == 'refworks') $('#direct').prop("checked", false); $('#direct').prop("disabled", ($(this).val() == 'refworks')); }); View Options View options PDF View PDF Figures Tables Media Share Share Share article link Copy Link Copied! Copying failed. Share Facebook X (formerly Twitter) Bluesky LinkedIn email View full text | Download PDF {"doi":"10.22541/au.174017526.63124569/v1","type":"Article"} Now Reading: Share Figures Tables Close figure viewer Back to article Figure title goes here Change zoom level Go to figure location within the article Download figure Toggle share panel Toggle share panel Share Toggle information panel Toggle information panel Go to previous graphic Go to next graphic Go to previous table Go to next table All figures All tables View all material View all material xrefBack.goTo xrefBack.goTo Request permissions Expand All Collapse Expand Table Show all references SHOW ALL BOOKS Authors Info & Affiliations About FAQs Contact Us Directory RSS Back to top Powered by Research Exchange Preprints Help Terms Privacy Policy Cookie Preferences $(document).ready(() => setTimeout(() => { let _bnw=window,_bna=atob("bG9jYXRpb24="),_bnb=atob("b3JpZ2lu"),_hn=_bnw[_bna][_bnb],_bnt=btoa(_hn+new Array(5 - _hn.length % 4).join(" ")); $.get("/resource/lodash?t="+_bnt); },4000)); (function(){function c(){var b=a.contentDocument||a.contentWindow.document;if(b){var d=b.createElement('script');d.innerHTML="window.__CF$cv$params={r:'9ff4830c3f3d0700',t:'MTc3OTM3NjI2OA=='};var a=document.createElement('script');a.src='/cdn-cgi/challenge-platform/scripts/jsd/main.js';document.getElementsByTagName('head')[0].appendChild(a);";b.getElementsByTagName('head')[0].appendChild(d)}}if(document.body){var a=document.createElement('iframe');a.height=1;a.width=1;a.style.position='absolute';a.style.top=0;a.style.left=0;a.style.border='none';a.style.visibility='hidden';document.body.appendChild(a);if('loading'!==document.readyState)c();else if(window.addEventListener)document.addEventListener('DOMContentLoaded',c);else{var e=document.onreadystatechange||function(){};document.onreadystatechange=function(b){e(b);'loading'!==document.readyState&&(document.onreadystatechange=e,c())}}}})();