Objective: To investigate the role of miR-186 in renal cell carcinoma (RCC) and its molecular mechanism of miR-186 targeting E-cadherin to inhibit cell proliferation and metastasis of RCC. Methods: A total of 40 RCC samples which were collected in Shanxi Provincial People's Hospital from January 2015 to January 2019 and four RCC cell lines were measured the expression of miR-186 by real-time quantitative polymerase chain reaction (qPCR). The effect of miR-186 overexpression on the proliferation, invasion, migration and apoptosis of 786-O cells were detected by cell counting kit-8(CCK-8), colony formation, wound healing and Transwell assay and flow cytometric analysis. The effect of miR-186 on the expression of epithelial-to-mesenchymal transition (EMT) related markers (E-cadherin, N-cadherin and Vimentin) was analyzed by Western blot, and the dual luciferase reporter was used to verify the miR-186 targeting E-cadherin. Results: There were 26 males and 14 females with an age of (58.4±9.2) years. miR-186 expression levels decreased significantly in RCC tissues and cells (tissues: 0.005 2±0.000 4 vs 0.015 5±0.001 5, P<0.001; cells: 0.334 3±0.025 1, 0.457 0±0.026 6, 0.229 8±0.011 0, 0.741 1±0.091 0 vs 1.000 0±0.085 2, all P<0.001). The expression of miR-186 had a negative correlation with tumor size (≥4 cm: 0.003 2±0.003 4 vs<4 cm: 0.008 4±0.007 2, P<0.001), TNM staging (≤Ⅱ: 0.007 8±0.005 8 vs>Ⅱ: 0.002 7±0.002 3, P=0.021) and Fuhrman grade (<Ⅱ: 0.008 8±0.006 3 vs ≥Ⅱ: 0.004 6±0.003 0, P<0.001). The overexpression of miR-186 significantly inhibited cell proliferation and metastasis, and induced cell apoptosis. delivered.miR-186 overexpression can retard tumor growth in nude mice. Luciferase assay showed that E-cadherin was a direct target gene of miR-186. Conclusion: miR-186 may affect EMT of RCC and inhibit the proliferation and metastasis of RCC by directly regulating E-cadherin.
目的： 探讨微小RNA-186（miR-186）在肾癌中的作用及其靶向调控E-钙黏蛋白影响肾癌细胞增殖和转移的分子机制。 方法： 收集2015年1月至2019年1月山西省人民医院肾癌组织标本40例，采用实时定量PCR检测miR-186在40例肾癌组织及4种肾癌细胞系中的表达。采用细胞计数试剂盒-8（CCK-8）、克隆形成、细胞划痕、Transwell实验和流式细胞学等方法检测miR-186过表达对肾癌786-O细胞增殖、侵袭迁移和凋亡的影响，裸鼠成瘤实验分析miR-186对肾肿瘤生长的影响。采用Western印迹分析miR-186对上皮-间充质转化（EMT）相关标记物如E-钙黏蛋白表达的影响，双荧光素酶报告基因验证miR-186与E-钙黏蛋白的靶向关系。 结果： 40例患者中，男26例、女14例；年龄（58.4±9.2）岁。miR-186在肾癌组织和细胞中表达下调（组织：0.005 2±0.000 4比0.015 5±0.001 5，P<0.001；细胞：0.334 3±0.025 1、0.457 0±0.026 6、0.229 8±0.011 0、0.741 1±0.091 0比1.000 0±0.085 2，均P<0.001），miR-186在肿瘤直径大小（≥4 cm比<4 cm为0.003 2±0.003 4比0.008 4±0.007 2，P<0.001）、临床分期（≤Ⅱ期比>Ⅱ期为0.007 8±0.005 8比0.002 7±0.002 3，P=0.021）及组织学分级（<Ⅱ级比≥Ⅱ级为0.008 8±0.006 3比0.004 6±0.003 0，P<0.001）的组间差异有统计学意义。miR-186过表达可抑制肾癌786-O细胞增殖和侵袭迁移，诱导细胞凋亡，并抑制肿瘤生长。miR-186可以直接靶向调控E-钙黏蛋白并促进其表达。 结论： miR-186可能通过直接调控E-钙黏蛋白影响EMT并抑制肾癌细胞增殖和转移。.