miR-29c-5p负性调控CPEB4对肾透明细胞癌细胞增殖、迁移的影响

The effect of miR-29c-5p on the proliferation and migration of clear cell renal carcinoma cells by negatively regulating CPEB4

目的探讨miR-29c-5p对胞质多聚腺苷酸化原件结合蛋白4(CPEB4)的负性调控作用,以及对肾透明细胞癌Caki-1细胞增殖、迁移的影响。方法采用Lipofectamine TM 2000将miR-NC(miR-NC组)、miR-29c-5p mimics(miR-29c-5p mimics组)、miR-29c-5p inhibitor(miR-29c-5p inhibitor组)、miR-29c-5p inhibitor+si-CPEB4转染(si-CPEB4+miR-29c-5p inhibitor组)转染至Caki-1细胞。采用MTT法和Transwell实验检测各组细胞增殖和迁移活性的影响,采用实时荧光定量PCR(QPCR)和Western blotting检测各组miR-29c-5p、CPEB4 mRNA和蛋白的表达水平。结果miR-29c-5p mimics组细胞的增殖和迁移活性明显低于miR-NC组(P<0.05);而miR-29c-5p inhibitor组细胞的增殖和迁移活性明显高于miR-NC组(P<0.05)。miR-29c-5p mimics组CPEB4 mRNA和蛋白的表达水平明显低于miR-NC组和miR-29c-5p inhibitor组,差异具有统计学意义(P<0.05);miR-29c-5p inhibitor组CPEB4 mRNA和蛋白的表达量明显高于miR-NC组和miR-29c-5p mimics组,差异具有统计学意义(P<0.05)。沉默CPEB4基因表达后,miR-29c-5p inhibitor组miR-29c-5p表达水平明显上调,差异具有统计学意义(P<0.05)。与miR-29c-5p inhibitor组比较,si-CPEB4+miR-29c-5p inhibitor组细胞的增殖和迁移活性明显降低,差异具有统计学意义(P<0.05)。结论miR-29c-5p可负性调控CPEB4的表达,从而抑制肾透明细胞癌的增殖和迁移活性。

Objective To investigate the negative regulation of miR-29c-5p on cytoplasmic polyadenylation primitive binding protein 4(CPEB4)and its effect on the proliferation and migration of renal clear cell carcinoma Caki-1 cells.Methods miR-NC(miR-NC group),miR-29c-5p mimics(miR-29c-5p mimics group),miR-29c-5p inhibitor(miR-29c-5p inhibitor group)and miR-29c-5p inhibitor+si-CPEB4(si-CPEB4+miR-29c-5p inhibitor group)were transfected into Caki-1 cells by Lipofectamine TM 2000.MTT assay and Transwell assay were used to detect the effects of cell proliferation and migration activity in each group.Real-time fluorescence quantitative PCR(QPCR)and Western blotting were used to detect the expression levels of miR-29c-5p,CPEB4 mRNA and CPEB4 protein in each group.Results The proliferation and migration activity of the cells in the miR-29c-5p mimics group was significantly lower than that in the miR-NC group(P<0.05);while the proliferation and migration activity of cells in the miR-29c-5p inhibitor group was significantly higher than that in the miR-NC group(P<0.05).The proliferation and migration activity of cells in the miR-29c-5p inhibitor group was significantly higher than that in the miR-NC group(P<0.05).The expression levels of CPEB4 in the miR-29c-5p mimics group were significantly lower than those in the miR-NC group and the miR-29c-5p inhibitor group(P<0.05).The expression levels of CPEB4 in the miR-29c-5p inhibitor group were significantly higher than those in the miR-NC group and the miR-29c-5p mimics group(P<0.05).After silencing the expression of CPEB4 gene,the expression of miR-29c-5p of miR-29c-5p inhibitor group increased significantly(P<0.05).Compared with the miR-29c-5p inhibitor group,the proliferation and migration activity of cells in the si-CPEB4+miR-29c-5p inhibitor group decreased significantly(P<0.05).Conclusion There are significant evidences that miR-29c-5p could inhibit the proliferation of clear cell renal cell carcinoma by down-regulation CPEB4 expression.