1.Expression of Exogenous r-Interferon Genes in Human HepatoceUuIar Carcinoma Cells Following Retroviral Transduction
Shubing QIAN ; Tengfei ZHANG ; Liang HU
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Retroviral vector pLXSN was employed to introduce human r-Interferon (IFN-?) gene into four human hepa-tocellular carcinoma cell lines (HCC). The G418-resistant colonies were isolated and cloned. PCR and RT-PCR analysis indicated that the integration and expression of IFN-? gene was shown only in the transduced cells. Using a bioassay method, we found that all genetically modified HCC cells can secrete varied amount of IFN-?. The results of flow cytome-try showed that the cell surface expression of HLA class I molecules significandy increased following transduction. Moreover , we firsdy indicated that the increase in the expression of one specific HLA class I antigen, HLA-A2, was almost in the same magnitude as that of the total HLA class I molecules after transduction with IFN-r gene.
2.Enhanced expression of HLA class Ⅰ molecules in human hepatocellular carcinoma cells transduced with γ-interferon gene
Shubing QIAN ; Tengfei ZHANG ; Shishu CHEN
Chinese Medical Journal 1998;111(4):319-322
Objective To investigate the expression of exogenous γ-interferon gene in human hepatocellular carcinoma cells following retroviral transduction and the effect on the expression of surface HLA class Ⅰ molecules.Methods Retroviral vector pLXSN was used to introduce human γ-interferon (IFN-γ) gene into four different human hepatocellular carcinoma cell lines (HCC). The G418-resistant colonies were isolated and cloned. The integration and expression of IFN-γ gene were determined by PCR and RT-PCR analysis. A bioassay method was used to test the amount of IFN-γ secreted by gene modified HCC cells. The expression of HLA class Ⅰ molecules in HCC cells were analyzed by flow cytometry using indirect fluorescence staining.Results Four different HCC cell lines were successfully transduced with human IFN-γ gene using retroviral vector. The integration and expression of IFN-γ gene were shown only in the transduced cells. All four genetically modified HCC cells can secrete varied amount of IFN-γ and demonstrate a significant up-regulation of surface HLA class Ⅰ antigens. One specific HLA class Ⅰ antigen, HLA-A2, has almost the same degree of increase as that of the total HLA class Ⅰ molecules after transduction with IFN-γ gene.Conclusions Gene modification with IFN-γ gene can significantly enhance the expression of HLA class Ⅰ molecules in HCC cells and may increase its immunogenicity. These gene modified tumor vaccines can be helpful in tumor biotherapy.