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Objective:To study the antitumor effect and mechanism of cocultured CIK cells modified with IL-24 gene and autologous DCs on HL-60 cells in vitro.Methods:DCs and CIK cells were prepared routinely from human peripheral blood mononuclear cells ( PBMC).IL-24 gene was transferred into CIK cells via electroporation.The cells obtained were named CIK-IL24.RT-PCR and ELISA were used to evaluate expression of IL-24 gene in transfected CIK cells.The phenotypic changes of CIK cells were identified by flowcytometry analysis.The concentration of IFN-γ and TNF-α in supernatant of CIK was determined by ELISA.FCM was used to determine the cytotoxicity of cocultured CIK cells modified with IL-24 gene and autologous DCs against HL-60 cells.Results:Eukaryotic expressing plasmid pcDNA3.0-IL24 was transferred into CIK cells successfully via electroporation.The expressing rate of CD3~+、CD3~+CD56~+ cells had no significant change in CIK cells.However,the rate of CD4~+CD25~+ cells was significantly decreased compared with that of the control group.Expression of adhesion molecules CD54,CXCR4 were significantly increased on CD3+CD56+ cells.CIK-IL24 cells produced markedly higher levels of IFN-γ and TNF-α as compared with the CIK cells.By comparison with non-transfected CIK cells co-cultured with DCs,transfected CIK cells co-cultured with DCs had a significantly higher lytic activity against HL-60 cells.Conclusion:IL-24 gene modification can enhance the anti-tumoral immunity of CIK cells,the mechanism of which might be related to the increased secretion of IFN-γ,TNF-α,up-regulation of adhesion molecule expression,and reduction of the rate of CD4~+CD25~+ cells in CIK cells.
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Objective: To evaluate the apoptosis-inducing effect of HPV-16 E6 siRNA associated with hIL-24 gene on human cervical cancer Ca Ski cells. Methods: HPV-16 E6 siRNA and hIL-24 gene eukaryotic expression vector alone or in combination were transfected into human cervical cancer Ca Ski cells. The expression level of HPV-16 E6 mRNA was detected by RT-PCR. The expression of tumor suppressor protein p53 was analyzed by western blot. The apoptosis of Ca Ski cells was analyzed by flow cytometry analysis. Results: The expression level of HPV-16 E6 mRNA decreased in Ca Ski cells in all the three groups after transfection with HPV-16 E6 siRNA and hIL-24 gene eukaryotic expression vector. The decrease was significant in co-transfection group ( P < 0. 05 ). The expression of tumor suppressor protein p53 and the apoptosis rate of Ca Ski cells increased in the three groups and the increase was significant in co-transfection group ( P < 0.05 ). Conclusion: Co-transfection of Ca Ski cells with HPV - 16 E6 siRNA and hIL-24 gene inhibits the expression of tumor gene HPV E6, recovers the activity of tumor suppressor protein p53, and induces the apoptosis of Ca Ski cells. HPV - 16 E6 siRNA associated with hIL-24 gene significantly increases the apoptotic rate of Ca Ski cells. They have synergistic effects.