构建可诱导表达MST1基因的慢病毒载体及其在肝癌细胞系Huh-7中的表达

Construction of inducible lentiviral expression vector containing MST1 gene and its expression in Huh-7 cell line

目的构建可诱导表达MST1基因的慢病毒载体,为进一步研究MST1与肝癌之间的关系提供理想的模型。方法克隆MST1基因到慢病毒载体pLVPT-tTRKRAB中,2种慢病毒载体(pLVPT-tTRKRAB-MST1和pLV-tTRKRAB-Red)与包装质粒通过293FT细胞系介导,分别包装成病毒颗粒,先后2次去感染肝癌细胞系Huh-7。培养7 d后,使用强力霉素(doxcycline)进行诱导,用Western blot检测MST1表达情况。结果构建了可诱导MST1真核表达病毒载体并获得相应的病毒,病毒可以高效感染目的肝癌细胞系Huh-7。在强力霉素诱导条件下,肝癌细胞系Huh-7可以表达MST1。结论成功构建出可诱导表达MST1基因的慢病毒载体,并获得在强力霉素诱导条件下可以表达MST1的肝癌细胞亚系。

Objective To construct inducible lentiviral vector containing MST1 gene,which provides an ideal model for further study of the relationship between liver cancer and MST1 gene. Methods We cloned MST1 into inducible lentiviral vector.Two lentiviral vectors（pLVPT-tTRKRAB-MST1 and pLV-tTRKRAB-Red） with package plasmids were contransfected into 293FT respectively,and the lentiviral viruses were harvested from 293FT.Viruses were used to infect liver cancer cell line（Huh-7） in tandem.We used doxcycline to induce the expression of target gene MST1 which was indentified by Western blot after 7-day cell culture. Results The recombinant inducable lentiviral vector containing MST1 gene was successfully constructed.The lentiviruses were also obtained and mediated by 293FT,which were highly efficient to infect liver cell lines Huh-7.The expression of MST1 was identified under Dox induction. Conclusions The recombinant inducable lentiviral vector containing MST1 gene has been successfully constructed.It is viable to obtain inducible cell lines Huh-7 with MST1 gene expression under Dox induction.