Raji细胞Id4基因甲基化检测及As2O3对其甲基化影响的研究  

Methylation of Id4 gene and inhibitive effect of arsenic trioxide on it in Raji cells

作　　者：曲凡[1] 

机构地区：[1]河北医科大学第四医院儿科,石家庄050011

出　　处：《中华血液学杂志》2010年第12期821-825,共5页Chinese Journal of Hematology

基　　金：河北省卫生厅医学科学研究重点课题计划（20090493）

摘　　要：目的 研究淋巴瘤细胞DNA结合抑制因子4（inhibitor of DNA binding,Id4）基因甲基化及As2O3去甲基化效应.方法 体外培养人Burkitt＇s淋巴瘤细胞系Raji细胞,设对照组和用不同浓度As2O3处理Raji细胞不同时间的实验组.用甲基化特异性聚合酶链反应（MS-PCR）检测各组Raji细胞Id4基因甲基化程度;逆转录聚合酶链反应（RT-PCR）检测不同浓度As2O3处理前后Raji细胞Id4mRNA的表达情况.应用MTT比色法检测细胞增殖情况;流式细胞术（FCM）检测细胞凋亡并分析As2O3对细胞周期的影响.结果 ①Raji细胞Id4基因呈完全甲基化状态,Raji细胞Id4基因高甲基化状态可被As2O3逆转,且与As2O3的剂量有关.②不同浓度As2O3处理Raji细胞48 h后,随着药物浓度增加,Raji细胞中Id4基因mRNA的相对表达量逐渐增加,呈一定剂量依赖关系.③不同浓度As2O3处理Raji细胞24、48、72 h后细胞增殖抑制率为12.15%～92.17%,且作用呈时间剂量依赖性（P＜0.05）;④FCM检测细胞凋亡结果显示,As2O3处理Raji细胞48 h后均出现明显的凋亡峰,对照组未见凋亡峰或仅有低平的凋亡峰;⑤FCM细胞周期分析结果显示,不同浓度As2O3处理Raji细胞48 h后,细胞阻滞于G0/G1期,且随着As2O3浓度的增加,阻滞作用增强,呈一定剂量依赖关系.结论 ①人Burkitt＇s淋巴瘤细胞系Raji细胞中Id4基因呈完全甲基化状态.②As2O3可以逆转Raji细胞Id4基因的高甲基化状态,使Id4 mRNA重新表达.③Raji细胞Id4基因呈高甲基化状态是其恶性增殖的原因之一.Objective To study methylation of Id4 gene and demethylation effect of arsenic trioxide （ATO） in Raji cells. Methods Human Burkitt＇s Raji lymphoma cells were cultared and treated with ATO at different concentrations and different time points. Methylated degree of Id4 gene was detected by methylation specificity polymerase chain reaction （ MS-PCR）, Id4 mRNA expression in Raji cell by reverse transcription polymerase chain reaction （RT-PCR）, the growth of cell by MTT assay, and cell apoptosis and cycle dispression in Raji cells treated with ATO for 48 h increased gradually with ATO concentration increasing in exATO treatment for 48 h in experiment group, the cells were arrested at G0/G1 phase in a dose-dependent manner. Conclusion Id4 gene presents exhaustive methylation in Raji cells. ATO can reverse the hypermethylation of Id4 gene and recover the expression of Id4 mRNA. Hypermethylation of ld4 gene is one of the reasons of Raji cells malignant proliferations.

关 键 词：淋巴瘤 砷剂 DNA甲基化 基因 Id4 聚合酶链反应 

分 类 号：R686[医药卫生—骨科学]