微小RNA-186-5p对肾癌细胞增殖与侵袭能力的影响

鲁广建

doi:10.16098/j.issn.0529-1356.2020.06.011

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解剖学报 ›› 2020, Vol. 51 ›› Issue (6) : 877-881. DOI: 10.16098/j.issn.0529-1356.2020.06.011

肿瘤生物学

微小RNA-186-5p对肾癌细胞增殖与侵袭能力的影响

鲁广建¹ 狄文玉² 张群妹³焦路阳^1*

作者信息 +

Effects of microRNA-186-5p on the proliferation and invasion ability of renal cancer cells

LU Guang-jian¹ DI We-yu² ZHANG Qun-mei³ JIAO Lu-yang^1*

Author information +

文章历史 +

摘要

目的观察微小RNA-186-5p（MiR-186-5p）对肾癌细胞侵袭和增殖的影响并探讨可能存在的机制。方法培养正常肾小管上皮细胞系HK-2、原代及转染MiR-186-5p的肾癌细胞（Caki-2细胞），Real-time PCR检测MiR-186-5p的水平，细胞计数试剂盒8（CCK-8）法评估各组细胞的存活率，Transwell法检测细胞的迁移和侵袭能力，Western blotting检测磷酸化蛋白激酶B（p-Akt）和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）水平。结果肾小管上皮细胞系Hk-2和转染了MiR186-5p的Caki-2细胞的MiR186-5p水平均显著高于Caki-2细胞（P<0.05）。HK-2细胞和Caki-2细胞的存活率接近100%，而转染了MiR-186-5p的Caki-2细胞的存活率显著降低至72.86%（P<0.05）。与HK-2细胞的迁移数目相比，Caki-2细胞迁移较多（P<0-05）；转染MiR-186-5p的Caki-2细胞迁移较少（P<0.05）。与HK-2细胞内Akt和mTOR分子磷酸化水平相比，Caki-2细胞内Akt和mTOR分子磷酸化水平显著增加（P<0.05）；而转染了MiR-186-5p的Caki-2细胞内Akt和mTOR分子的磷酸化水平有所下降（P<0.05）。结论 MiR-186-5p能够有效抑制肾癌细胞的侵袭与增殖，可能与其抑制Akt/mTOR信号通路的激活相关。

Abstract

Objective To observe the effect of microRNA-186-5p（MiR-186-5p）on the invasion and proliferation of renal cancer cells in vitro and explore the possible mechanisms. Methods Culture normal renal tubular epithelial cell line HK-2, primary and MiR-186-5p-transfected renal cancer cells (Caki-2 cells). Real-time PCR was used to detect the level of MiR-186-5p, the cell counting kit-8 (CCK-8) method was used to evaluate the survival rate of each group of cells, the Transwell method was used to detect the cell migration and invasion ability, and Western blotting was used to detect phosphorylated protein kinase B (p-Akt) and phosphorylated mammalian rapamycin target protein (p-mTOR) levels. Results The renal tubular epithelial cell line HK-2 and MiR-186-5p transfected Caki-2 cells had significantly higher MiR-186-5p levels than Caki-2 cells (P<0.05). The survival rates of HK-2 cells and Caki-2 cells were close to 100%, while the survival rate of Caki-2 cells transfected with MiR-186-5p. was significantly reduced to 72.86% (P<0.05). Compared with the migration number of HK-2 cells, Caki-2 cells migrated more (P<0.05); MiR-186-5p transfected Caki-2 cells migrated less (P<0.05). Compared with the phosphorylation level of Akt and mTOR molecules in HK-2 cells, the phosphorylation level of Akt and mTOR molecules in Caki-2 cells was significantly increased (P<0.05); Caki-2 cells transfected with MiR-186-5p. The phosphorylation level of Akt and mTOR molecules decreased (P<0.05). Conclusion MiR-186-5p can effectively inhibit the invasion and proliferation of renal cancer cells, which may be closely related by inhibiting the activation of Akt/mTOR signaling pathway.

导出引用

鲁广建狄文玉张群妹焦路阳. 微小RNA-186-5p对肾癌细胞增殖与侵袭能力的影响[J]. 解剖学报. 2020, 51(6): 877-881 https://doi.org/10.16098/j.issn.0529-1356.2020.06.011

LU Guang-jian DI We-yu ZHANG Qun-mei JIAO Lu-yang. Effects of microRNA-186-5p on the proliferation and invasion ability of renal cancer cells[J]. Acta Anatomica Sinica. 2020, 51(6): 877-881 https://doi.org/10.16098/j.issn.0529-1356.2020.06.011

中图分类号： R73.37

参考文献

［1］ Rini BI, Hutson TE, Figlin RA, et al. Sunitinib in patients with metastatic renal cell carcinoma: clinical outcome according to international metastatic renal cell carcinoma database consortium risk group［J］. Clin Genitourin Cancer, 2018, 16(4):298-304.

［2］ Motzer RJ, Tannir NM, McDermott DF, et al. Nivolumab plus ipilimumab versus sunitinib in advanced renal-cell carcinoma［J］. N Engl J Med, 2018, 378(14):1277-1290.

［3］ Stephen S. Paraneoplastic cough and renal cell carcinoma［J］. Can Respir J, 2016, 2016:5938536.

［4］ Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019［J］. CA Cancer J Clin, 2019, 69(1):7-34.

［5］ Liu Y, Pan Y, Zheng W, et al. MiR-186-5p is highly expressed in alcohol-induced cardiomyocytes and regulates apoptosis through the target gene XIAP ［J］. Chinese Journal of Bioengineering, 2019 (5): 53-62.（in Chinese）

刘叶, 潘玥, 郑魏, 等. mi-186-5p在酒精诱导的心肌细胞中高表达并通过靶基因XIAP调控细胞凋亡水平［J］. 中国生物工程杂志, 2019(5):53-62.

［6］ Luo HSh, Wan YY, Tian X, et al. Expression and mechanism of miR-20a-5p in pancreatic cancer ［J］. Journal of Gastroenterology and Hepatology, 2019,28(3):335-340.（in Chinese）

罗和生, 万一圆, 田霞, 等. miR-20a-5p在胰腺癌中的表达及作用机制研究［J］. 胃肠病学和肝病学杂志, 2019,28(3):335-340.

［7］ Sun YG, Wang ZhY, Yang YL, et al. MiR-186-5p regulates the reverse mechanism of RAB2A in breast cancer cell doxorubicin resistance by targeting ［J］. Chinese Pharmacology Bulletin, 2018, 34 (12): 45-50.（in Chinese）

孙玉国, 王照岩, 杨玉玲, 等. MiR1865p通过靶向调控RAB2A在乳腺癌细胞阿霉素耐药性中的逆转作用机制［J］. 中国药理学通报, 2018, 34(12):4550.

［8］ Zhang GX, Xu XW, Meng B, et al. MiR-186-5p inhibits epithelial-mesenchymal transition of lung adenocarcinoma cells through targeted regulation of PTTG1 ［J］. Chinese Journal of Biochemistry and Molecular Biology, 2017, (4): 67-72.（in Chinese）

张国新, 徐新伟, 孟斌, 等. miR-186-5p通过靶向调控PTTG1抑制肺腺癌细胞的上皮间质转化［J］. 中国生物化学与分子生物学报, 2017,(4):67-72.

［9］ Wang PY, Tian JT, Feng YK, et al. Research progress of the role of MiRNA in reverse cholesterol transport ［J］. Chinese Journal of Atherosclerosis, 2018, 26 (10): 1052-1056.（in Chinese）

王鹏宇, 田江天, 冯玉宽,等. MiRNA在胆固醇逆向转运中的作用研究进展［J］. 中国动脉硬化杂志, 2018, 26(10):1052-1056.

［10］ Jiang WJ, Fang M, Su JW, et al. Effects and mechanism of MicroRNA-486 and TGF-β1 on calcification of human aortic valve ［J］. China Journal of Modern Medicine, 2018, 28 (32): 31-38.（in Chinese）

蒋伟坚, 方明, 苏锦文,等. MicroRNA-486和TGF-β1对人主动脉瓣膜钙化的影响及其机制研究［J］. 中国现代医学杂志, 2018, 28(32):31-38.

［11］ Zhang GX, Xu XW, Meng B, et al. MiR-186-5p inhibits epithelial-mesenchymal transition of lung adenocarcinoma cells through targeted regulation of PTTG1 ［J］. Chinese Journal of Biochemistry and Molecular Biology, 2017, (4): 380-385.（in Chinese）

张国新, 徐新伟, 孟斌,等. MiR-186-5p通过靶向调控PTTG1抑制肺腺癌细胞的上皮-间质转化［J］. 中国生物化学与分子生物学报, 2017, (4):380-385.

［12］ Song D, Wang ZY, Huang P. Expression and role of microRNA-186 in human hepatocellular carcinoma ［J］. Cancer, 2017, 37 (7): 750-761.（in Chinese）

宋丹, 王孜尧, 黄平. 微RNA-186在人肝细胞肝癌中的表达及作用［J］. 肿瘤, 2017, 37(7):750-761.

［13］ Wang ShX, Zhang ZJ, Yang ShF, et al. Clinical significance of MiRNA expression differences in human kidney cancer cell lines ［J］. Chinese Journal of Cancer Prevention and Treatment, 2018, 25 (3): 175-179.（in Chinese）

王世先, 张遵俊, 杨水法,等. 人肾癌细胞株MiRNA表达差异临床意义研究［J］. 中华肿瘤防治杂志, 2018, 25(3):175-179.

［14］ Xu LW. The role of MiR-1865p in the invasion, metastasis and angiogenesis of renal clear cell carcinoma ［D］. Zhejiang University, 2016: 1-138.（in Chinese）

许力为.MiR-186-5p在肾透明细胞癌侵袭转移和血管生成中作用的研究［D］.浙江大学,2016:1-138.

［15］ Ma JH. Abnormal mTOR signaling pathway of renal cancer cells and the mechanism of action of everolimus ［J］. Chinese Journal of Urology, 2013, 34 (3): 229-231.（in Chinese）

马建辉. 肾癌细胞mTOR信号通路异常与依维莫司作用机制［J］. 中华泌尿外科杂志, 2013, 34(3):229-231.

［16］ Guo ShSh, Zhang ZhY, Wang Y. Effects of PI3K/Akt/MTOR signaling pathway on proliferation, migration and invasion of renal cancer A498 cells ［J］. Oncology Progress, 2017, 15(12): 1412-1416.（in Chinese）

郭双双, 张治业, 王颖. PI3K/Akt/MTOR信号通路对肾癌A498细胞增殖、迁移及侵袭的影响研究［J］. 癌症进展, 2017, 15(12): 1412-1416.