Inhibition of the proliferation of hepatocellular carcinoma cells by miR-214 via regulation of E2F3 expression / 中国比较医学杂志

ABSTRACT

Objective To explore the effect of inhibition of miR-214 expression on the proliferation of hepatocellular carcinoma cells via regulation of E2F3 expression.Methods The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was examined by RT-PCR.Hepatocellular carcinoma cells were transfected with miR-214 NC and miR-214 mimics with liposomes.The expression of miR-214 was detected by RT-PCR.The cell viability was detected by MTT assay.Cell apoptosis was detected by Hoechst staining.Cell cycle was detected by flow cytometry.Western blot, RT-PCR and dual luciferase reporter gene assay were used to detect whether E2F3 was a downstream target gene of miR-214.Results The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was 0.83±0.08, 0.32±0.03, 0.33±0.03, and 0.08±0.01, respectively.The expression of miR-214 in the HepG2 cells was the lowest, so HepG2 cells were selected as the subsequent experimental cell line.Compared with the miR-214 NC group, the expression of miR-214 (0.65±0.06 vs.0.14±0.01) was up-regulated, the cell viability (0.35±0.03 vs.0.69±0.06) was decreased, cell apoptosis rate [(36.37±3.43)% vs.(3.74±0.34)%] was increased, the G1 phase of the cell cycle (57.79±5.78 vs.45.319±4.53) was prolonged, the expression of E2F3 protein (0.23±0.02 vs.0.98±0.09) and mRNA (0.24±0.02 vs.0.99±0.10) was significantly down-regulated in the miR-214 mimics group (P<0.01).Conclusion miR-214 mimics suppress the HepG2 cell proliferation via targeted down-regulation of E2F3 expression.