Li Wen-ling, Yan Yun-li, Shan Bao-an, Zheng Li-fen, Li Qiao-xia, Zhou Na-jing
Cell Biology Division, Institute of Basic Medicine, Hebei Medical University, Shijiazhuang 050017, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep;20(5):522-5.
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.
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.
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.
Exogenous IL-18 gene can inhibit the proliferation and in-vivo tumorigenicity of C6 cells.
建立表达白细胞介素 -18（IL -18）基因的大鼠C6胶质瘤细胞系C6/IL -18，并探讨IL -18对C6细胞生长的影响。
通过逆转录病毒载体将IL -18基因导入C6细胞。经G418筛选后，获得C6/IL -18细胞。采用逆转录 -聚合酶链反应（RT -PCR）检测C6/IL -18细胞中IL -18 mRNA的表达。通过流式细胞术和免疫细胞化学染色检测IL -18蛋白的表达。为分析所表达的IL -18蛋白的活性，采用酶联免疫吸附测定（ELISA）检测用C6/IL -18细胞培养上清诱导大鼠脾细胞分泌γ-干扰素（IFN -γ）的能力。采用噻唑蓝（MTT）比色法和流式细胞术检测C6/IL -18细胞的体外增殖情况。利用大鼠模型观察C6/IL -l8细胞的致瘤活性。
IL -18 mRNA和蛋白在C6/IL -18细胞中稳定表达。C6/IL -18细胞的培养上清可诱导大鼠脾细胞分泌IFN -γ。同时，与亲代C6细胞相比，C6/IL -18细胞的增殖率和体内致瘤性明显降低。
外源性IL -18基因可抑制C6细胞的增殖和体内致瘤性。
