逆转录病毒介导IL-18基因在大鼠胶质瘤细胞C6中的表达

目的 :建立表达IL 18基因的大鼠胶质瘤细胞C6 /IL 18,并探讨外源性IL 18基因对C6细胞生长的影响。方法 :应用逆转录病毒载体 ,将IL 18基因导入C6细胞。经G4 18筛选后 ,获得表达IL 18分子的细胞克隆C6 /IL 18。用RT PCR法检测目的基因mRNA的表达 ;用流式细胞和免疫细胞化学染色法检测目的蛋白的表达。用ELISA法检测C6 /IL 18细胞培养上清诱导脾细胞分泌IFN γ的能力 ,以确定IL 18的生物学活性。用MTT比色法检测细胞体外增殖的状况 ,用流式细胞技术检测细胞的增殖活性 ,并建立大鼠胶质瘤模型 ,观察C6 /IL 18细胞体内致瘤性的改变。结果 :外源IL 18基因于mRNA水平和蛋白水平可获得稳定表达 ,并可诱生大鼠脾细胞分泌IFN γ。同时 ,该细胞系的体外增殖率及体内致瘤性 ,均较亲代C6胶质瘤细胞明显下降。结论 :外源性IL 18基因能部分地抑制C6细胞的体外增殖率和体内致瘤性。建立了可进一步用于相关肿瘤基因免疫和基因治疗研究的大鼠胶质瘤细胞系C6 /IL 18。

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-gamma 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 were stably expressed in C6/IL-18 cells. The culture supernatant of C6/IL-18 cells induced secretion of IFN-gamma 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.