过表达miR-185通过靶向DNA甲基转移酶1抑制胃癌细胞增殖和转移

Overexpressed miR-185 inhibits gastric cancer proliferation and metastasis via targeting DNA methyltransferase

目的探讨miR-185对胃癌细胞中的抑制作用及可能机制。方法Real time PCR检测miR-185在患者胃癌组织、癌旁组织以及胃癌细胞株中的表达情况;并在胃癌细胞系中过表达miR-185后,CCK-8法检测细胞增殖;Transwell法检测细胞迁移能力;生物信息学网站预测miR-185的靶点,并用荧光素酶报告基因实验验证miR-185的靶基因,用Western blot检测miR-185转染胃癌细胞系后靶蛋白的表达。结果miR-185在胃癌组织表达水平低于癌旁组织,在胃癌细胞系HGC-27和MGC-803中的表达水平低于胃上皮黏膜细胞GES-1。过表达miR-185能够抑制胃癌细胞的增殖和迁移。DNA甲基转移酶1(DNMT1)是miR-185直接作用靶点,miR-185过表达能够直接下调胃癌细胞中DNMT1的表达,通过抑制DNMT1表达发挥抑制胃癌细胞增殖及迁移的作用。结论过表达miR-185抑制胃癌细胞的增殖和迁移,通过靶向抑制DNMT1蛋白发挥作用。

Objective To investigate the inhibitory effect and mechanism of miR-185 in gastric cancer cells.Methods Real-time PCR was used to detect the expression of miR-185 in gastric cancer,adjacent tissues,and gastric cancer cell lines.After miR-185 transfection in cells,CCK-8 was used to detect the cell proliferation;Transwell assay was used to detect the cell migration ability.The target of miR-185 was predicted by bioinformatics website and verified by luciferase reporter gene experiment.Western blot was used to confirm the expression of target protein in miR-185 transfected cells.Results miR-185 expression levels were lower in gastric cancer tissue than in adjacent normal tissue,and lower in HGC-27 and MGC-803 cell lines than in gastric epithelial mucosa GES-1 cells.Transfection of miR-185 inhibited the proliferation and migration of gastric cancer HGC-27 cells,DNA methyltransferase 1(DNMT1)is a direct target of miR-185,overexpressed MiR-185 directly downregulated DNMT1 expression level,furthermore to inhibit the proliferation and migration of gastric cancer HGC-27.Conclusion The overexpression of miR-185 inhibits the proliferation and migration of gastric cancer cells via targeting DNMT1.