Wang Y B, Zhang Z L, Shao J K, Li R S
Department of Urology, Shanxi Provincial People's Hospital, Taiyuan 030012, China.
Department of Breast Surgery, Shanxi Provincial People's Hospital, Taiyuan 030012, China.
Zhonghua Yi Xue Za Zhi. 2021 Apr 13;101(14):1020-1025. doi: 10.3760/cma.j.cn112137-20210110-00068.
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. 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. 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, <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 <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, <0.001), TNM staging (≤Ⅱ: 0.007 8±0.005 8 vs>Ⅱ: 0.002 7±0.002 3, =0.021) and Fuhrman grade (<Ⅱ: 0.008 8±0.006 3 vs ≥Ⅱ: 0.004 6±0.003 0, <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. miR-186 may affect EMT of RCC and inhibit the proliferation and metastasis of RCC by directly regulating E-cadherin.
探讨miR-186在肾细胞癌（RCC）中的作用及其靶向E-钙黏蛋白抑制RCC细胞增殖和转移的分子机制。收集2015年1月至2019年1月在山西省人民医院采集的40例RCC样本及4种RCC细胞系，采用实时定量聚合酶链反应（qPCR）检测miR-186的表达。通过细胞计数试剂盒-8（CCK-8）、集落形成、伤口愈合、Transwell实验及流式细胞术分析检测miR-186过表达对786-O细胞增殖、侵袭、迁移及凋亡的影响。采用蛋白质免疫印迹法分析miR-186对上皮-间质转化（EMT）相关标志物（E-钙黏蛋白、N-钙黏蛋白和波形蛋白）表达的影响，并采用双荧光素酶报告基因验证miR-186靶向E-钙黏蛋白。患者共40例，男性26例，女性14例，年龄（58.4±9.2）岁。RCC组织和细胞中miR-186表达水平显著降低（组织：0.005 2±0.000 4对0.015 5±0.001 5，<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对1.000 0±0.085 2，均<0.001）。miR-186的表达与肿瘤大小（≥4 cm：0.003 2±0.003 4对<4 cm：0.008 4±0.007 2，<0.001）、TNM分期（≤Ⅱ期：0.007 8±0.005 8对>Ⅱ期：0.002 7±0.002 3，=0.021）及Fuhrman分级（<Ⅱ级：0.008 8±0.006 3对≥Ⅱ级：0.004 6±0.003 0，<小于0.001）呈负相关。miR-186过表达显著抑制细胞增殖和转移，并诱导细胞凋亡。miR-186过表达可抑制裸鼠肿瘤生长。荧光素酶实验表明E-钙黏蛋白是miR-186的直接靶基因。miR-186可能通过直接调控E-钙黏蛋白影响RCC的EMT，进而抑制RCC的增殖和转移。
