【摘要】 目的:本研究拟从分子水平探讨IL-18基因转染对C6胶质瘤细胞周期及其相关基因表达的影响。方法:用流式细胞仪检测C6/IL-18细胞周期、细胞增殖指数的变化;以MTT法检测细胞的增殖活性及细胞抑制率,用RT-PCR、Western blot分析C6/IL-18细胞PCNAc、yclin D1c、yclin B1、p21 mRNA及蛋白的表达。结果:与C6/LXSN细胞及亲代C6细胞相比,C6/IL-18细胞周期有所改变,表现为G0/G1期细胞增多、G2/M期细胞减少,细胞增殖指数(PI)与细胞增殖活性降低。C6/IL-18细胞的PCNAc、yclin D1c、yclin B1 mRNA及蛋白表达降低,p21 mRNA及蛋白表达增高。结论:外源性IL-18基因表达可降低C6细胞的增殖活性,下调PCNA,cyclin D1,cyclin B1表达,上调p21基因表达,其作用机制有待进一步研究。更多还原

【Abstract】 Objective:The purpose of this study was to investigate the transfection of interleukin-18（IL-18） gene on the cell cycle and related gene expression of C6 glioma cells.Methods:The distribution of cell cycle and change in proliferation index（PI） were detected by flow cytometry.The proliferation of C6 glioma cells and the inhibitory effect of IL-18 gene transfection on C6 glioma cells were assessed by MTT assay.Reversed transcriptase-polymerase chain reaction and Western blotting analysis were applied to test the mRNA and protein expressions of proliferating cell nuclear antigen（PCNA）,cyclin D1,cyclin B1,p21 in C6/IL-18,C6/LXSN,and C6 cells,respectively.Results: Compared with C6/LXSN and parent C6 cells,the proportion of C6/IL-18 cells arrested at G₀/G₁ increased significantly but the proportion of C6/IL-18 cells at G₂/M phase decreased.The PI and the proliferation ability of C6/IL-18 cells were lower than that of C6/LXSN and C6 cells.Both the mRNA and protein level of PCNA,cyclin D1,and cyclin B1 of C6/IL-18 cells was down-regulated,while the mRNA and protein level of p21 was up-regulated.Conclusions:Exogenous IL-18 gene transfection could reduce the proliferation ability of C6 cells,down-regulate the expression of PCNA,cyclin D1,and cyclin B1,and up-regulate the expression of p21.The mechanism needs to be further studied.更多还原