COMMD7 gene promotes HepG2 proliferation and migration by activiating the PI3K/Akt signal pathway / 局解手术学杂志

ABSTRACT

Objective To investigate the mechanism of COMMD7,a hepatocellular carcinoma gene,promoting human hepatocellular carcinoma cell line (HePG2 cells)proliferation and migration.Methods The specific siRNA (small interference RNA)was designed for COMMD7 gene,siRNA transfected HepG2 cells was set as Si-HePG2 group,and the PTEN inhibitor treating group (Si +BpV-HePG2),the control group (HePG2),the empty vector group (HePG2 was infected empty vector,N-HePG2 group)were also set up.qRT-PCR was per-formed to evaluate the mRNA expression changes of COMMD7 and PTEN,and Western blot was performed to test the expression changes of COMMD7,PTEN,p-AKT,and AKT.MSP method was performed to detect methylation level.CCK8 was performed to test cell proliferation a-bility.Transwell was performed to detect the invasion of cells.Results The results of qRT-PCR showed that the expression of COMMD7 gene in Si-HePG2 group was lower (0.101 times)than the control group and the N-HePG2 group,and the expression of PTEN gene was higher (2.841 times)than the control group and the N-HePG2 group,and all the results above were of statistically singificant difference (P <0.05). The results of Western blot showed that in Si-HePG2 group,the expression of COMMD7 reduced obviously,the expression of PTEN increased and the expression of p-AKT was significantly inhibited.In Si +Bpv-HePG2 group,the expression of PTEN was significantly inhibited and the expression of p-AKT was increased.The result of MSP showed that compared with the control group and the N-HePG2 group,Si-HePG2 group was more obviously demethylated,which indecated that the expression of COMMD7 gene could induce PTEN demethylation.The results of CCK8 showed that the proliferation in Si-HePG2 group was decreased compared with the control group,N-HePG2 group and Si +BpV-HePG2 group,and the difference was singificant (P <0.05).The results of Transwell showed that the numbers of cell permeating septum in Si-HePG2 group,N-HePG2 group,control group and PTEN group were (17.4 ±2.7),(36.2 ±3.2),(41.6 ±4.5)and (47.6 ±1.8)respec-tively,which were obviously decreased with a significant difference (P <0.05).Conclusion siRNA interference decreased the expression of COMMD7.It can induced the demethylation of PTEN gene in HePG2 and increase the expression of PTEN gene to inhibit PI3K/AKT signaling pathway,thus decreasing the proliferation,migration and invasion ability of HePG2 cells.