miR-182-5p靶向SMAD4对前列腺癌PC-3细胞增殖的影响

Effects of miR-182-5p targeting SMAD4 on proliferation of prostate cancer PC-3 cells

目的探讨miR-182-5p对前列腺癌PC-3细胞增殖的影响及可能机制。方法将前列腺癌PC-3细胞分为四组,分为空白组(未进行转染)、miR-182-5p阴性对照组(转染无义序列)、miR-182-5p模拟物组(转染miR-182-5p模拟物)、miR-182-5p抑制物组(转染miR-182-5p抑制物),采用实时定量PCR验证转染效率,CCK-8法检测各组PC-3细胞的增殖能力,Western blot检测各组PC-3细胞SMAD4蛋白表达,应用生物信息学网站分析SMAD4是否为miR-182-5p的靶基因,应用双荧光素酶报告基因实验验证。结果CCK-8实验结果显示,miR-182-5p过表达后,PC-3细胞增殖能力增强,SMAD4蛋白表达下降;抑制miR-182-5p表达后,PC-3细胞增殖能力降低,SMAD4蛋白表达升高。生物信息学分析结果显示SMAD4基因3’UTR区存在miR-182-5p的结合位点,双荧光素酶实验证实了SMAD4为miR-182-5p的靶基因。结论miR-182-5p可能通过靶向SMAD4调控前列腺癌PC-3细胞的增殖。

Objective To investigate the effect and mechanism of miR-182-5 p on the proliferation of prostate cancer PC-3 cells.Methods The PC-3 cells were divided into blank control group(not transfected),miR-182-5 p negative control group(transfected unrelated sequence),miR-182-5 p mimics group(transfected miR-182-5 p mimics),and miR-182-5 p inhibitor group(transfected miR-182-5 p inhibitor).Real time PCR method was used to verify the transfection efficiency.The proliferation of PC-3 cells was detected by CCK-8,the expression of SMAD4 protein was detected by Western blot.Bioinformatics and dual luciferase reporter gene experiment was used to verify whether SMAD4 is a target gene of miR-182-5 p.Results The result of CCK-8 showed that after over expression of miR-182-5 p,the proliferation ability of PC-3 cells was increased and the expression of SMAD4 protein was decreased;after inhibition of miR-182-5 p expression,the proliferation ability of PC-3 cells was decreased and the expression of SMAD4 protein was increased.Bioinformatics analysis showed that there was a binding site of miR-182-5 p in the 3’UTR region of SMAS4 gene.Conclusion miR-182-5 p may regulate PC-3 cell proliferation by targeting SMAD4.