Retrovirus-mediated IL-18 gene expression in rat glioma cell C6

AIM: To set up rat C6 glioma cell line C6/IL-18 expressing IL-18 gene and explore the effect of IL-18 on the growth of C6 cells. METHODS: The IL-18 gene was transferred into the C6 cells by a retrovirus vector. After screening with G418, the C6/IL-18 cells were obtained. IL-18 mRNA expression in C6/IL-18 cells was detected with RT-PCR. The expression of IL-18 protein was detected by flow cytometry and immunocytochemical staining. In order to analyze the activity of the expressed IL-18 protein, ELISA was used to detect the ability to secrete IFN-γ by rat splenocytes induced with the culture supernatant of C6/IL-18 cells. The in vitro proliferation of C6/IL-18 cells was detected by MTT colorimetry and flow cytometry. The rat model was used to observe the tumorigenic activity of the C6/IL-18 cells. RESULTS: IL-18 mRNA and protein stably expressed in C6/IL-18 cells. The culture supernatant of C6/IL-18 cells induced secretion of IFN-γ by rat splenocytes. At the same time, the proliferation rate and in vivo tumorigenicity of C6/IL-18 cells were markedly reduced as compared with parental C6 cells. CONCLUSION: Exogenous IL-18 gene can inhibit the proliferation and in vivo tumorigenicity of C6 cells.