论文部分内容阅读
目的:检测4种人脑成胶质细胞瘤细胞株中成视网膜细胞瘤蛋白结合锌指结构基因1(retinoblastoma protein-interacting zinc finger gene1,RIZ1)基因启动子区的甲基化状态,进一步认识RIZ1在胶质瘤发病机制中的作用。方法:应用甲基化特异性PCR法(methylation specific PCR,MSP)检测4种人脑成胶质细胞瘤细胞株U87、U251、A172和T98中RIZ1基因启动子区的甲基化水平,其中U87细胞发生甲基化,被选为后续实验对象。RT-PCR检测U87细胞经5-Aza-CdR处理前后RIZ1 mRNA表达量的变化,MTT检测5-Aza-CdR对U87细胞生长增殖的影响。结果:4种人脑成胶质细胞瘤细胞株中U87和U251细胞中检测到RIZ1基因启动子区域发生甲基化;U87细胞经5-Aza-CdR处理后其RIZ1 mRNA的表达量上调;MTT法检测显示5-Aza-CdR能抑制U87的生长增殖,且与5-Aza-CdR的浓度和作用时间呈负相关。结论:RIZ1基因启动子高甲基化是人脑成胶质细胞瘤细胞株中RIZ1基因表达下调的重要机制。
AIM: To detect the methylation status of the promoter region of retinoblastoma protein-interacting zinc finger gene 1 (RIZ1) in four human glioblastoma cell lines, and to further understand the relationship between RIZ1 Role in the pathogenesis of glioma. Methods: Methylation-specific PCR (MSP) was used to detect the methylation levels of RIZ1 promoter in U87, U251, A172 and T98 cell lines of human glioblastoma. U87 Cells were methylated and selected as follow-up subjects. The changes of RIZ1 mRNA expression in U87 cells treated with 5-Aza-CdR were detected by RT-PCR. The effect of 5-Aza-CdR on the proliferation and proliferation of U87 cells was detected by MTT assay. Results: The methylation of RIZ1 promoter region was detected in U87 and U251 cells in 4 human glioblastoma cell lines. The expression of RIZ1 mRNA in U87 cells was up-regulated by 5-Aza-CdR treatment. MTT The results of the assay showed that 5-Aza-CdR inhibited the proliferation of U87 cells and was negatively correlated with the concentration and duration of 5-Aza-CdR. CONCLUSION: RIZ1 promoter hypermethylation is an important mechanism of down-regulation of RIZ1 gene expression in human glioblastoma cell lines.