Establishment of retroviral vector inducibly expressing RN181 and its expression in hepatocellular carcinoma / 中华肝胆外科杂志

ABSTRACT

Objective To study the expression and biological functions of RN181 in SMMC7721 cells,the retroviral vector was constructed.Methods Gene cloning techniques were used to construct pRetrox-TRE3G/RN1S1 recombinant vector.The regulating plasmid pRetroX-TRE3G/RN181 or the response plasmid of pRetroX-Tet3G were respectively cotansfected into GP2-293 cells with Envelope Vector plasmid to package retrovirus after routine identification.Both viruses co-infected target cells SMCC7721,and then were selected by G418 to obtain stable cell lines.The stable cell lines were induced by doxycycline (DOX),and then verified by RT-PCR and Western blotting.CCK-8 was used to evaluate the effect of RN181 on growth of SMMC7721 cells.Results We constructed the recombinant plasmid.Stable recombinant plasmid were verified by screening.And there were significant differences of RN181 between the induced and uninduced cell lines through RT-PCR and Western blot.Conclusions We have successfully constructed the inducible stable RN181 expression SMCC7721 cell,which can be used as an effective cell model to study the biological functions of RN181.We found RN181 could suppress the proliferation and invasion in SMMC7721 cells in vitro.