过表达miR-138-5p对胃癌MKN-45细胞增殖和侵袭的影响及可能机制

Effect of over-expression of miR-138-5p on proliferation and invasion of gastric cancer MKN-45 cells and its possible mechanism

目的分析过表达miR-138-5p对胃癌MKN-45细胞增殖、侵袭的影响及机制。方法体外培养胃癌MKN-45细胞,分为空白对照组(未转染)、miR-138-5p阴性对照组(转染miR-NC无意义序列)和miR-138-5p激动剂组(转染miR-138-5p模拟物)。实时荧光定量PCR检测各组细胞中miR-138-5p、KLF12的相对表达水平;Western blot检测各组细胞中KLF12蛋白表达;Targetscan和双荧光素酶实验分析KLF12与miR-138-5p的靶向关系;MTT法检测各组细胞增殖能力;Transwell实验检测各组细胞侵袭能力。结果miR-138-5p过表达使MKN-45细胞中KLF12蛋白表达下降。Targetscan网站分析及双荧光素酶实验证实KLF12为miR-138-5p的靶基因。MTT及Transwell实验结果显示,过表达miR-138-5p使MKN-45细胞增殖能力和侵袭能力下降。结论过表达miR-138-5p可以抑制胃癌MKN-45细胞增殖及侵袭能力,其机制可能与其靶向调控KLF12的表达有关。

Objective To investigate the effect and mechanism of over-expression of miR-138-5p on the proliferation and invasion of MKN-45 cells.Methods Gastric cancer MKN-45 cells were cultured in vitro and divided into blank control group(non transfected),miR-138-5p negative control group(transfected with miR-NC)and miR-138-5p agonist group(transfected with miR-138-5p mimics).The relative expression of miR-138-5p and KLF12 in each group were detected by qRT-PCR.The expression of KLF12 was detected by Western blot.The targeting relationship between KLF12 and miR-138-5p was analyzed by targetscan and double luciferase experiment.The proliferation of MKN-45 cells was detected by MTT.The invasion of MKN-45 cells was detected by Transwell.Results Overexpression of miR-138-5p decreased the expression of KLF12 in MKN-45 cells.Targetscan website analysis and double luciferase experiment confirmed that KLF12 was the target gene of miR-138-5p.MTT and Transwell experiments showed that over-expression of miR-138-5p could inhibit the proliferation and invasion of MKN-45 cells.Conclusion Overexpression of miR-138-5p could inhibit the proliferation and invasion of MKN-45 cells by targeting KLF12.