miR-195-5p通过靶向ROCK1抑制肾癌细胞迁移、侵袭和上皮-间质转化

miR-195-5p inhibits renal cell carcinoma cells migration,invasion and epithelial to mesenchymal transition by targeting ROCK1

目的探讨miR-195-5p对肾癌细胞迁移、侵袭和上皮-间质转化的影响。方法通过转染miR-195-5p mimics或inhibitors分别过表达或抑制肾癌细胞中miR-195-5p的表达,转染靶向Rho相关螺旋蛋白激酶1(ROCK1)的小干扰RNA敲低肾癌细胞中ROCK1的表达量,利用细胞划痕实验和Transwell小室实验分别检测肾癌细胞的迁移和侵袭能力。通过双荧光素酶报告实验验证miR-195-5p对ROCK1的靶向调控作用,利用免疫印迹试验检测ROCK1及上皮-间质转化相关蛋白的表达水平。结果过表达miR-195-5p可显著抑制肾癌细胞的迁移、侵袭和上皮-间质转化,而抑制miR-195-5p的表达可明显促进肾癌细胞的迁移、侵袭和上皮-间质转化(P<0.05)。miR-195-5p可通过靶向ROCK1调控其在肾癌细胞中表达。敲低ROCK1后可部分抵消miR-195-5p inhibitors对肾癌细胞迁移、侵袭和上皮-间质转化的影响。结论miR-195-5p可通过靶向ROCK1抑制肾癌细胞的迁移、侵袭和上皮-间质转化。

Objective To investigate the effects of miR-195-5p on the migration,invasion and epithelial to mesenchymal transition(EMT)in renal cell carcinoma(RCC).Methods miR-195-5p mimics or inhibitors were transfected to overexpress or suppress miR-195-5p expression in RCC cells.Small interfering RNA targeting Rho-associated coiled-coil containing protein kinases 1(ROCK1)was transfected to knockdown ROCK1 in RCC cells.Would healing assay and Transwell invasion assay were used to detect the migration and invasion capacity of RCC cells,respectively.Dual-luciferase reporter assay was performed to verify that ROCK1 was a direct target of miR-195-5p.The protein expressions of ROCK1 and EMT markers were detected with Western blot.Results Overexpression of miR-195-5p significantly inhibited RCC cells migration,invasion and EMT,while suppression of miR-195-5p obviously facilitated RCC cells migration,invasion and EMT(P<0.05).miR-195-5p could directly target ROCK1 and regulate its expression in RCC cells.Knockdown of ROCK1 partly abolished the effects of miR-195-5p suppression on RCC cells migration,invasion and EMT.Conclusion miR-195-5p inhibits RCC cells migration,invasion and EMT by targeting ROCK1.