Effects of exogenous mitofusin-2 gene transfection on the biological behaviors of hepatic carcinoma cell in vitro

Objective To investigate the effects of mitofusin-2 gene(mfn2) on the biological behaviors of hepati carcinoma cell line HepG2 in vitro.Methods The recombinant eukaryotic expressing plasmidc pEGFPmfn2 was constructed,identified by using recombinant DNA technique,and then transfected into HepG2 cells.After the transfected cells were screened with G418,the mRNA expression of the cell was detected by RT-PCR,so was the protein expression by Western-blot.The proliferation of HepG2 was measured by MTT and apoptosis was assessed by Annexin-V /PI double staining flow cytometry.Results The plasmid pEGFPmfn2 was digested by restriction enzyme and electrophoresis of the products showed a mfn2 fragment of 2 300 bp and a pEGFP-N2 of 4 700 bp.The expressions of mfn2 mRNA and protein were detected by RT-PCR and Western-blot.Cell proliferation was inhibited following mfn2 transfection.Flow cytometry assay indicated that the apoptotic rate was increased in the transfected group,as compared with other groups.Conclusions The recombinant eukaryotic expressing plasmid pEGFPmfn2 has been successfully constructed.Exogenous mfn2 gene can inhibit the proliferation of hepatic carcinoma cell and induce apoptosis of the cell.