Journal of Shandong University (Health Sciences) ›› 2019, Vol. 57 ›› Issue (10): 74-79.doi: 10.6040/j.issn.1671-7554.0.2019.270

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siRNA-mediated silencing of RRM2 in the treatment of epithelial ovarian cancer xenograft in nude mice

XUE Ting1, WANG Liming1, JIAO Jinwen1, LI Yong1, GUO Ailian1, ZHAO Han2   

  1. 1. Department of Gynecology;
    2. Department of Pathology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong, China
  • Published:2022-09-27

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Abstract: Objective To explore the feasibility of siRNA silencing of ribonucleotide reductase M2(RRM2)gene combined with cisplatin on the treatment of human ovarian cancer subcutaneous xenograft in nude mice. Methods After conventional cultivation of human ovarian cancer cell line SKOV3 in vitro, subcutaneous injections of ovarian cancer were administered to nude mice to create a xenograft model. Twenty-four tumor-burdened mice were randomly divided into the control group(n=6), cisplatin group (n=6), siRNA group (n=6), and siRNA+cisplatin group (n=6). Tumor volume was measured and tumor growth inhibition rate was calculated. RRM2 gene expressions at mRNA level and protein level were detected by RT-PCR and immunohistochemistry. Results After single drug treatment, the tumor volumes of control group, siRNA group, cisplatin group and siRNA+cisplatin group were(358.28±46.40),(261.38±52.49),(261.65±31.97)and(189.37±41.67)mm3, respectively, with a significant statistical difference (FsiRNA-RRM2= 山 东 大 学 学 报 (医 学 版)57卷10期 -薛婷,等.siRNA介导RRM2基因沉默治疗人卵巢癌裸鼠移植瘤 \=-22.399,PsiRNA-RRM2<0.001; Fcisplatin=22.254, Pcisplatin<0.001). There was no interaction effect between the two drugs (F=0.474,P=0.499). The tumor growth inhibition rates of the four groups were 0, 36.39%, 41.10% and 64.33%, respectively. The single drug(siRNA-RRM2 or cisplatin)treatment reduced the expression levles of RRM2 mRNA(FsiRNA-RRM2=9.37, PsiRNA-RRM2=0.006; Fcisplatin=11.90, Pcisplatin=0.003)and protein (FsiRNA-RRM2=8.71, PsiRNA-RRM2=0.008; Fcisplatin=13.01, Pcisplatin=0.002), with a significant statistical difference. There was no interaction effect between the two drugs (FRRM2 mRNA=3.93, PRRM2 mRNA=0.061; FRRM2 protein=1.72, PRRM2 protein=0.204). Conclusion siRNA alone or combined with cisplatin can effectively inhibit the growth of human ovarian cancer subcutaneous xenograft, which may be related to the silencing of RRM2 gene expression. RRM2 gene silencing may be a potential treatment regimen for ovarian cancer in future.

Key words: Ribonucleotide reductase, siRNA/RNAi, Cisplatin, Ovarian cancer, Nude mice

CLC Number: 

  • R711.75
[1] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018[J]. CA Cancer J Clin, 2018, 68(1): 7-30.
[2] Xia G, Wang H, Song Z, et al. Gambogic acid sensitizes gemcitabine efficacy in pancreatic cancer by reducing the expression of ribonucleotide reductase subunit-M2(RRM2)[J]. J Exp Clin Cancer Res, 2017, 36(1): 107. doi: 10.1186/s13046-017-0579-0.
[3] Chen CW, Li Y, Hu S, et al. DHS(trans-4,4'-dihydroxystilbene)suppresses DNA replication and tumor growth by inhibiting RRM2(ribonucleotide reductase regulatory subunit M2)[J]. Oncogene, 2019, 38(13): 2364-2379.
[4] Li C, Zheng J, Chen S, et al. RRM2 promotes the progression of human glioblastoma[J]. J Cell Physiol, 2018, 233(10): 6759-6767.
[5] Zhang M, Wang J, Yao R, et al. Small interfering RNA(siRNA)-mediated silencing of the M2 subunit of ribonucleotide reductase: a novel therapeutic strategy in ovarian cancer[J]. Int J Gynecol Cancer, 2013, 23(4): 659-666.
[6] 何志军,陈先祥,蔡庆和,等.移植瘤体积不同计测方法的比较[J].中国比较医学杂志, 2009, 19(9): 47-50. HE Zhijun, CHEN Xianxiang, CAI Qinghe, et al. Comparative study of the different calculating and measuring methods in the implanted tumor volume[J]. Chinese Journal of Comaparative Medicine, 2009, 19(9): 47-50.
[7] Duroux-Richard I, Giovannangeli C, Apparailly F. CRISPR-Cas9: a revolution in genome editing in rheumatic diseases[J]. Joint Bone Spine, 2017, 84(1): 1-4.
[8] Wong HL, Shen Z, Lu Z, et al. Paclitaxel tumor-priming enhances siRNA delivery and transfection in 3-dimensional tumor cultures[J]. Mol Pharm, 2011, 8(3): 833-840.
[9] Wang J, Lu Z, Wang J, et al. Paclitaxel tumor priming promotes delivery and transfection of intravenous lipid-siRNA in pancreatic tumors[J]. J Control Release, 2015, 216: 103-110. doi: 10.1016/j.jconrel.2015.08.012.
[10] Sun TM, Du JZ, Yao YD, et al. Simultaneous delivery of siRNA and paclitaxel via a “two-in-one” micelleplex promotes synergistic tumor suppression[J]. ACS Nano, 2011, 5(2): 1483-1494.
[11] Hu Q, Li W, Hu X, et al. Synergistic treatment of ovarian cancer by co-delivery of survivin shRNA and paclitaxel via supramolecular micellar assembly[J]. Biomaterials, 2012, 33(27): 6580-6591.
[12] Wang L, Meng L, Wang XW, et al. Expression of RRM1 and RRM2 as a novel prognostic marker in advanced non-small cell lung cancer receiving chemotherapy[J]. Tumor Biology, 2014, 35(3): 1899-1906.
[13] Grolmusz VK, Karászi K, Micsik T, et al. Cell cycle dependent RRM2 may serve as proliferation marker and pharmaceutical target in adrenocortical cancer[J]. Am J Cancer Res, 2016, 6(9): 2041-2053.
[14] Putluri N, Maity S, Kommagani R, et al. Pathway-centric integrative analysis identifies RRM2 as a prognostic marker in breast cancer associated with poor survival and tamoxifen resistance[J]. Neoplasia, 2014, 16(5): 390-402.
[15] Zheng S, Wang X, Weng YH, et al. siRNA knockdown of RRM2 effectively suppressed pancreatic tumor growth alone or synergistically with doxorubicin[J]. Mol Ther Nucleic Acids, 2018, 12: 805-816. doi: 10.1016/j.omtn.2018.08.003.
[16] Wang A, Zhao C, Liu X, et al. Knockdown of TBRG4 affects tumorigenesis in human H1299 lung cancer cells by regulating DDIT3, CAV1 and RRM2[J]. Oncol Lett, 2018, 15(1): 121-128.
[17] Roberts CM, Shahin SA, Wen W, et al. Nanoparticle delivery of siRNA against TWIST to reduce drug resistance and tumor growth in ovarian cancer models[J]. Nanomedicine, 2017, 13(3): 965-976.
[18] Grunda JM, Fiveash J, Palmer CA, et al. Rationally designed pharmacogenomic treatment using concurrent capecitabine and radiotherapy for glioblastoma; gene expression profiles associated with outcome[J]. Clin Cancer Res, 2010, 16(10): 2890-2898.
[19] 张梦,王蕾,焦金文,等.siRNA介导RRM2沉默对卵巢癌细胞顺铂敏感性影响研究[J].中华肿瘤防治杂志, 2014, 21(4): 273-279. ZHANG Meng, WANG Lei, JIAO Jinwen, et al. Influence of small interference RNA-mediated RRM2 silence on the DDP sensitivity of ovarian cancer cell[J]. Chin J Cancer Prev Treat, 2014, 21(4): 273-279.
[20] 侯志斌,李菊英,苟笑丹. RRM2对人胃癌细胞MKN-45增殖、侵袭及顺铂耐药作用的研究[J].医学综述, 2018, 24(13): 194-198. HOU Zhibin, LI Juying, GOU Xiaodan. Study of RRM2 on proliferation, invasion and cisplatin resistance to MKN-45 cells[J]. Medical Recapitulate, 2018, 24(13): 194-198.
[21] Li J, Pang J, Liu Y, et al. Suppression of RRM2 inhibits cell proliferation, causes cell cycle arrest and promotes the apoptosis of human neuroblastoma cells and in human neuroblastoma RRM2 is suppressed following chemotherapy[J]. Oncol Rep, 2018, 40(1): 355-360.
[22] Che F, Du H, Zhang W, et al. MicroRNA-132 modifies angiogenesis in patients with ischemic cerebrovascular disease by suppressing the NFkappaB and VEGF pathway[J]. Mol Med Rep, 2018, 17(2): 2724-2730.
[23] Duxbury MS, Whang EE. RRM2 induces NF-kappaB-dependent MMP-9 activation and enhances cellular invasiveness[J]. Biochem Biophys Res Commun, 2007, 354(1): 190-196.
[24] Zhang W, Liu Y, Wang CW. S100A4 promotes squamous cell laryngeal cancer Hep-2 cell invasion via NF-kB/MMP-9 signal[J]. Eur Rev Med Pharmacol Sci, 2014, 18(9): 1361-1367.
[25] Yi C, Zhang L, Li L, et al. Establishment of an orthotopic transplantation tumor model in nude mice using a drug-resistant human ovarian cancer cell line with a high expression of c-Kit[J]. Oncol Lett, 2014, 8(6): 2611-2615.
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