Effects and mechanism of miR-214 on hepatocellular carcinoma

ABSTRACT

Objective:To explore the role of miR-214 in the progression of hepatocellular carcinoma (HCC) and its inhibitory mechanisms in depressing the signaling pathway ofβ-catenin, this study was conducted.Methods:We ectopically expressed miR-214 in HepG2 cells to obtain cell lines Lv-miR-214-HepG2 and their control Lv-control-HepG2. Differences between the two cell lines were compared in cell growth, proliferation, colony forming ability and cell cycles. RT-PCR method was applied for the quantification ofβ-catenin mRNA expression. Western-blot method was applied for the determination of the protein level ofβ-catenin and their downstream targets (ie. Cyclin D1, c-Myc and TCF-1). The effect of miR-214 on cells was further explored through RNA interference and restoring miR-214 expression.Results:In comparison with negative (Lv-control-HepG2) and blank (HepG2) control, a significant inhibition of cell growth and proliferation caused by miR-214 was observed after 48~72h of cell culture experiments (P0.05). By comparing to the RT-PCR and Western-blot results of control, the miR-214 treatment led to a slightly decrease in theβ-catenin mRNA expression (P>0.05), but an extremely inhibition in the protein level ofβ-catenin and its downstream targets Cyclin D1, c-Myc, and TCF-1 (P<0.05).Conclusions:miR-214 functions as a suppressor during the progression of HCC, and its inhibitory role was achieved by down-regulatingβ-catenin signaling pathway.