蛋白质精氨酸甲基转移酶5促进人卵巢癌HO8910细胞迁移

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摘要 目的: 探讨蛋白质精氨酸甲基转移酶5(protein arginine methyltransferase 5，PRMT5)对人卵巢癌HO8910细胞上皮间质转化(epithelial-to-mesenchymal transition，EMT)和迁移能力的影响。方法: 利用瞬时转染方法建立PRMT5过表达的HO8910细胞株，设空白对照组(野生型HO8910细胞株)、阴性对照组(转染pCMV-myc质粒)、实验组(转染pCMV-Myc-PRMT5质粒)；同时采用shRNA干扰方法建立诱导型稳定敲低PRMT5基因的HO8910细胞株，设pLKO对照组(感染空载体慢病毒)、pKLO+ Dox(100 ng/mL)组、shPRMT5组(感染PRMT5 shRNA 慢病毒)和shPRMT5+ Dox(100 ng/mL)组；通过免疫印迹和qRT-PCR实验检测PRMT5 mRNA表达和干扰效率。实验分为shPRMT5组、shPRMT5+Dox组、pCMV-Myc组、pCMV-Myc-PRMT5组；Transwell实验检测细胞迁移能力；免疫印迹法检测EMT相关蛋白表达。结果： 免疫印迹和qRT-PCR验证成功建立过表达PRMT5和稳定诱导型敲低PRMT5的卵巢癌HO8910细胞株。PRMT5敲低后，卵巢癌HO8910细胞迁移能力显著下降(P均<0.01)，间质标志相关分子包括基质金属蛋白酶2，波形蛋白，N-钙黏蛋白表达下调，上皮标志分子E-钙黏蛋白表达上调。此外，在卵巢癌细胞中外源表达PRMT5可显著促进HO8910细胞迁移(P均<0.05)。结论： PRMT5能够促进卵巢癌HO8910细胞EMT进程，促进细胞迁移。

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	赵康容
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关键词 ：蛋白质精氨酸甲基转移酶5, 上皮间质转化, 卵巢癌, 迁移

Abstract：Objective: To investigate the effect of protein arginine methyltransferase 5(PRMT5) on epithelial-to-mesenchymal transition(EMT) and migration ability in human ovarian cancer HO8910 cells. Methods: PRMT5 overexpressed HO8910 cell line was established by transient transfection method, and divided into blank control group (wild-type HO8910 cell line), negative control group (HO8910 cells transfected with pCMV-Myc plasmid), and experimental group (HO8910 cells transfected with pCMV-Myc-PRMT5 plasmid). At the same time, HO8910 cell lines with stable PRMT5 knockdown were established by shRNA interference and divided into pLKO control group (infected with empty lentivirus), pKLO+Dox group (100 ng/mL), shPRMT5 group (infected with PRMT5 shRNA lentivirus) and shPRMT5+Dox group (100 ng/mL). The mRNA expression and interference efficiency of PRMT5 were detected by Western blotting or qRT-PCR. The experiment was divided into shPRMT5 group, shPRMT5+Dox group, pCMV-Myc group and pCMV-Myc-PRMT5 group; Transwell assay was used to detect the cell migration ability. The expression of EMT-related proteins was detected by Western blotting. Results: Western blotting and qRT-PCR verify the establishment of ovarian cancer HO8910 cell lines with PRMT5 overexpression and stable knockdown PRMT5.After PRMT5 knockdown, the mobility of ovarian cancer HO8910 cells was significantly decreased (all P<0.01), and the expression of mesenchymal marker related molecules including matrix metalloproteinase-2, vimentin, and N-cadherin was down-regulated, and the expression of epithelial marker E-cadherin was up-regulated.In addition, exogenous PRMT5 expression in ovarian cancer cells greatly up-regulated the migration ability of HO8910 cells (all P<0.05). Conclusion: PRMT5 could promote the EMT process of ovarian cancer HO8910 cells and enhance cell migration.

收稿日期: 2020-03-24

基金资助: 国家自然科学基金资助项目(81170573)

通讯作者: 邵根宝(通讯作者)，教授，硕士生导师，E-mail：gbshao07@ujs.edu.cn

作者简介: 赵康容(1993—)，女，硕士研究生

引用本文:

赵康容，孙爱琴，林琼，邵根宝. 蛋白质精氨酸甲基转移酶5促进人卵巢癌HO8910细胞迁移[J]. 江苏大学学报:医学版, 2020, 30(05): 430-434. ZHAO Kang-rong, SUN Ai-qin, LIN Qiong, SHAO Gen-bao.

Protein arginine methyltransferase 5 promotes the migration of human ovarian cancer HO8910 cells

. Journal of Jiangsu University(Medicine Edition), 2020, 30(05): 430-434.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2020/V30/I05/430

［1］Siegel RL, Miller KD, Jemal A. Cancer statistics\[J\]. CA Cancer J Clin, 2018, 68(1):7-30.

［2］Momenimovahed Z， Tiznobaik A， Taheri S， et al. Ovarian cancer in the world: epidemiology and risk factors\[J\]. Int J Womens Health, 2019, 11: 287-299.

［3］Bedford MT, Clarke SG. Protein arginine methylation in mammals: who, what, and why\[J\]. Molecular Cell, 2009, 33(1): 1-13.

［4］Ichikawa T, Shanab O, Nakahata S, et al. Novel PRMT5mediated arginine methylations of HSP90A are essential for maintenance of HSP90A function in NDRG2low ATL and various cancer cells\[J\]. Biochim Biophys Acta Mol Cell Res, 2020, 1867(2):118615.

［5］Stopa N, Krebs JE, Shechter D. The PRMT5 arginine methyltransferase: many roles in development, cancer and beyond\[J\]. Cell Mol Life Sci, 2015, 72(11):2041-2059.

［6］Karkhanis V, Alinari L, Ozer HG, et al. Protein arginine methyltransferase 5 represses tumor suppressor miRNAs that downregulate CYCLIN D1 and cMYC expression in aggressive Bcell lymphoma\[J\]. J Biol Chem, 2020, 295(5):1165-1180.

［7］Holmes B, BenavidesSerrato A, Saunders JT, et al. The protein arginine methyltransferase PRMT5 confers therapeutic resistance to mTOR inhibition in glioblastoma\[J\]. J Neurooncol, 2019, 145(1):11-22.

［8］Vinet M, Suresh S, Maire V, et al. Protein arginine methyltransferase 5: A novel therapeutic target for triplenegative breast cancers\[J\]. Cancer Med, 2019, 8(5):2414-2428.

［9］Radzisheuskaya A, Shliaha PV, Grinev V, et al. PRMT5 methylome profiling uncovers a direct link to splicing regulation in acute myeloid leukemia\[J\]. Nat Struct Mol Biol, 2019, 26(11):999-1012.

［10］魏野, 王冉冉, 韩田田, 等. 蛋白质精氨酸甲基转移酶5对人卵巢癌HO8910细胞增殖能力的影响\[J\]. 医学研究生学报, 2018, 31(6):565-572.

［11］Shao G, Wang J, Li Y, et al. Lysinespecific demethylase 1 mediates epidermal growth factor signaling to promote cell migration in ovarian cancer cells\[J\]. Sci Rep, 2015, 5:15344.

［12］Deng X, Shao G, Zhang HT, et al. Protein arginine methyltransferase 5 functions as an epigenetic activator of the androgen receptor to promote prostate cancer cell growth\[J\]. Oncogene, 2017, 36(9): 1223-1231.

［13］Li Y, Wan X, Wei Y, et al. LSD1mediated epigenetic modification contributes to ovarian cancer cell migration and invasion\[J\]. Oncol Rep, 2016, 35(6): 3586-3592.

［14］Shao G, Wang R, Sun A, et al. The E3 ubiquitin ligase NEDD4 mediates cell migration signaling of EGFR in lung cancer cells\[J\]. Mol Cancer, 2018, 17(1): 24.

［15］Shailesh H, Zakaria ZZ, Baiocchi R, et al. Protein arginine methyltransferase 5 (PRMT5) dysregulation in cancer\[J\]. Oncotarget, 2018, 9(94):36705-36718.

［16］Zhao T, Jiang W, Wang X, et al. ESE3 inhibits pancreatic cancer metastasis by upregulating Ecadherin\[J\]. Cancer Res, 2017, 77(4):874-885.