Overexpression of miR-34c-5p inhibits invasion and migration of gastric cancer cells / 中国药理学通报

ABSTRACT

Aim To study the expression of microRNA(miR-34c-5p)in gastric cancer tissues and the mechanism of regulating the invasion and migration of gastric cancer cells. Methods The expression of miR-34c-5p mRNA in human gastric cancer tissues and cells was detected by quantitative real-time polymerase chain reaction(RT-qPCR). An overexpression model of miR-34c-5p was constructed in human gastric cancer cells. The transfection of lentivirus was observed under inverted fluorescence microscope, and the transfection efficiency was verified by RT-qPCR. Scratch test and Transwell assay were used to detect the effects of overexpression of miR-34c-5p on invasion and migration of gastric cancer cells. Western blot was used to detect the expression of MAP2K1, E-Cadherin and Vimentin in gastric cancer cells of each group. Dual-luciferase reporter assay was employed to detect whether MAP2K1 was a target gene of miR-34c-5p. Results The expression of miR-34c-5p mRNA in 10 human gastric cancer tissues was significantly lower than that in adjacent tissues(P<0.05). The expression of miR-34c-5p mRNA in gastric cancer HGC-27 cell was significantly lower than that in normal gastric mucosa GES-1 cell(P<0.01). Compared with blank control group and miR-34c-5p-NC group, miR-34c-5pmimics group significantly increased its miR-34c-5p mRNA expression level(P<0.05). The results of scratch test and Transwell experiment showed that overexpression of miR-34c-5p significantly inhibited the migration and invasion ability of gastric cancer cells(P<0.05, P<0.01). Western blot results showed that the protein expression levels of MAP2K1 and Vimentin significantly decreased and E-Cadherin markedly increased in gastric cancer HGC-27 cells after overexpression of miR-34c-5p(P<0.05). In addition, compared with mimic-NC+miR-34c-5p-mimics group, the lucifase activity of pmiR-MAP2K1-WT plasmid decreased(P<0.01), while the lucifase activity of pmiR-MAP2K1-MUT plasmid was not changed. Conclusions The low expression of miR-34c-5p in gastric cancer tissues and cells may inhibit the invasion and migration of gastric cancer cells through targeted regulation of MAP2K1 expression.