miR-30b-5p靶向调节Snail蛋白抑制胃癌增殖、侵袭、转移和上皮-间质转化

miR-30b-5p targets Snail to inhibit proliferation,invasion,metastasis and epithelial-mesenchymal transformation of gastric cancer

目的探讨miR-30b-5p在胃癌组织中的表达及对胃癌细胞增殖、侵袭、转移的影响。方法RT-qPCR检测胃癌患者癌、癌旁组织、胃癌细胞系SGC-7901、BGC-823、AGS和HGC-27以及正常胃上皮细胞系GES中miR-30b-5p的水平;在HGC-27细胞中转染miR-30b-5p mimics后,CCK-8法测定细胞增殖情况;Transwell小室法测定细胞侵袭情况;划痕法测定细胞迁移情况;Western blot检测细胞中E-cadherin,N-cadherin以及Vimentin蛋白的表达;Targetscan.org网站预测miR-30b-5p的靶基因,荧光素酶报告基因实验对靶基因进行验证;Western blot检测靶蛋白和下游调控蛋白的表达情况。结果miR-30b-5p在癌组织中表达显著低于癌旁组;在多株胃癌细胞中的表达显著低于胃上皮细胞,其中HGC-27表达最低。HGC-27细胞转染miR-30b-5pmimics后,细胞增殖、侵袭和迁移能力受到抑制,同时E-cadherin表达上调,N-cadherin表达下调,Vimentin表达下调。miR-30b-5p能够与Snail蛋白的3′UTR区域靶向结合,miR-30b-5p的水平与Snail蛋白表达负相关。结论miR-30b-5p在胃癌组织和细胞中表达下调,miR-30b-5p过表达能抑制胃癌细胞的增殖、迁移、侵袭以及EMT。

Objective To investigate the expression of miR-30b-5p in gastric cancer tissues and its effects on proliferation,invasion and metastasis of gastric cancer cells.Methods The levels of miR-30b-5p in gastric cancer patients,adjacent tissues,gastric cancer cell lines SGC-7901,BGC-823,AGS and HGC-27,and normal gastric epithelial cell lines GES were detected by RT-qPCR.After transfection of miR-30b-5p mimics into HGC-27 cells,proliferation of cells was measured by CCK-8 method.Invasion of cells was deter mined by Transwell assay.Migration of cells was measured by scratch method.The expressions of E-cadherin,N-cadherin and Vimentin were detected by Western blot.Target genes of miR-30b-5p were predicted by Targetscan.org,and the target genes were verified by luciferase reporter gene assay.The expressions of target protein and downstream regulatory protein were detected by Western blot.Results The expression of miR-30b-5p in cancer tissue was significantly lower than that in paracancer group.The expression of HGC-27 in gastric cancer cells was significantly lower than that in gastric epithelial cells,and the expression of HGC-27 was lowest.After transfection of miR-30b-5p mimics in HGC-27 cells,proliferation,invasion and migration of cells were inhibited,while the expression of E-cadherin was up-regulated,the expression of N-cadherin was down-regulated,and the expression of Vimentin was down-regulated.miR-30b-5p can bind to the 3′UTR region of Snail protein,and the level of miR-30b-5p is negatively correlated with Snail protein expression.Conclusion The expression of miR-30b-5p was down-regulated in gastric cancer tissues and cells,and overexpression of miR-30b-5p could inhibit the proliferation,migration,invasion and EMT of gastric cancer cells.