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miR-875-5p inhibits the proliferation, migration and invasion of gastric cancer cells by targeting USF2
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Abstract:
Objective: To investigate the effects of miR-875-5p on proliferation, migration and invasion of gastric cancer (GC) cells and its mechanism. Methods: The expression level of miR-875-5p was detected by qPCR in GC cell lines (BGC-823, HGC-27,MGC-803, SGC-7901, AGS, MKN-45) and gastric epithelial GES-1 cells. MiR-875-5p mimics/inhibitors or their negative control plasmids (miR-NC/anti-miR-NC) were transfected into AGS or MKN-45 cells by liposome transfection technique to construct miR-875-5p overexpression or knockdown cell model. Cells in blank control group (Control group) were not transfected. The effects of miR-875-5p on cell proliferation, clone formation, migration and invasion were detected by CCK-8 assay, colony formation assay and Transwell assay, respectively. The targeting relationship between miR-875-5p and upstream stimulatory factor 2 (USF2) was verified by dual-luciferase reporter gene assay. The regulation of miR-875-5p on USF2 as well as the expression of USF2 protein was confirmed by WB assay. After the construction of MKN-45 cell transplanted tumor model in nude mice, the effect of miR-875-5p overexpression on tumorigenesis of MKN-45 cells was detected. Results: The expression level of miR-875-5p in 6 GC cells was significantly lower than that in gastric epithelial GES-1 cells (all P<0.01). Compared with Control group and miR-NC group, the proliferation, clone formation rate, number of migrated and invaded cells, and USF2 protein expression level in AGS cells were significantly decreased in miR-875-5p mimic group (P<0.05 or P<0.01), while those were significantly increased in MKN-45 cells of miR-875-5p inhibitor group (P<0.05 or P<0.01). Dual-luciferase reporter gene assay demonstrated that miR-875-5p could directly target USF2 gene. In vivo tumorigenesis experiment results showed that overexpression of miR-875-5p significantly inhibited the growth of MKN-45 cell transplanted tumors (all P<0.01). Conclusion: miR-875-5p inhibits proliferation, migration and invasion of GC cells by targeting USF2.
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