In vitro experimental study on impacts of miR-214 on the proliferation of human hepatocellular carcinoma

Abstract

Objective To investigate the impact of micro ribonucleic acid -214 (miR-214) on the proliferation of hepatocellular carcinoma (HCC) and its mechanism. Methods MHCC97L cells were respectively transfected using miR-214 mimics and negative control mimics to establish M214 group, negative control (NC214) group. And untransfected control (Ctrl) group was established. The contents of miR-214 of MHCC97L cells in three groups were detected by fluorescence quantitative polymerase chain reaction (PCR). Cell counting kit (CCK)-8 assay was used to define the cell proliferation inhibition rate. Plate cloning formation assay was used to define the cell clonality. Cell apoptosis rate was detected by flow cytometery. The expressions of protein c-myc, Bax and B-cell lymphoma (Bcl)-2 were detected by Western blot assay. The comparison of three groups was conducted using one way analysis of variance and pairwise comparison using LSD-t test. Results The miR-214 content of MHCC97L cells in M214 group was (2 536±7) times of that in Ctrl group, where significant difference was observed (LSD-t=58.75, P<0.05). The cell proliferation inhibition rates were (15.33±0.62) %, (24.07±0.75) %, (41.02±0.91) % at the 24, 48, 72 h in M214 group, and significant difference was observed compared with that in Ctrl group (LSD-t =14.64, 19.87, 31.86; P<0.05). The clone formation quantity was (5.3±0.7) /well in M214 group, which was significantly lower compared with that in Ctrl group [(37.1±1.0)/well] (LSD-t =-12.51, P<0.05). The cell apoptosis rate was (42.1±3.2)% in M214 group, which was significantly higher compared with that in Ctrl group [(7.0±0.7) % ] (LSD-t=35.66, P<0.05). In M214 group, the expressions of protein c-myc, Bcl-2 decreased and Bax increased. Conclusion The miR-214 can inhibit the proliferation of human HCC cells through down-regulating the protein c-myc expression and Bcl-2/Bax ratio. Key words: Carcinoma, hepatocellular; MicroRNAs; Cell proliferation; Apoptosis