Retrovirus mediated stable expression of human GW112 gene in human gastric cancer cells SGC-7901

Objective To construct pLEGFP-N1-GW112 retrovirus expression vector and increase the expression of human GW112 gene in human gastric cancer SGC-7901 cells with retrovirus-mediated method.Methods The retrovirus vector pLEGFP-N1-GW112 containing signal peptide sequence was constructed by DNA recombinant technology and then transfected into RetroPacked PT67 cell line with Lipofectamine 2000.The viral supernatants were obtained from the PT67 clones and used to infect SGC-7901 cells.SGC-7901-GW112 and control SGC-7901-GFP cells were established after selected by G418.Real Time-PCR were used to analyze GW112 mRNA expression.Results The recombinant retrovirus vector pLEGFP-N1-GW112 containing signal peptide was successfully constructed,meanwhile virus package cell line were established and the high titer recombined retrovirus virus were obtained from PT67 clones and infected SGC-7901 cell successfully.Real-Time PCR results revealed GW112 mRNA levels were increased 4.2 fold in SGC-7901-GW112 cell compaired with its control.Conclusion SGC-7901-GW112 cells stablly overexpressing GW112 and SGC-7901-GFP were established,which may provide good basic for functional research of GW112 in gastric cancer.