Abstract:Objective:To investigate the feasibility of constructing hepatocellular carcinoma cell strain in which mitofusin-2 gene is stably expressed.
Methods :Mitofusin-2 gene was inserted into the eukaryotic expressing vector pEGFP-N2 to construct expressing plasmid pEGFPmfn2. pEGFPmfn2 was transfected into hepatocellular carcinoma cell strain HepG2 by Lipofectamine 2000 and was selected with G418 for 30 d. The mRNA expression was detected by RT-PCR and protein expression by Western-blot.
Results:(1) pEGFPmfn2 expression plasmid was constructed succesfully; (2) pEGFPmfn2 was transfected into cultured HepG2 succesfully and positive cell clone was obtained; (3) mitofusin-2 gene was stably expressed in transfected HepG2/mfn2.
Conclusions:Hepatocellular carcinoma cell strain HepG2 with stable mitofusin-2 gene expression was successfully constructed, which could be applied to explore the role of mitofusin-2 gene in the genesis and development of hepatic carcinoma.