As_2O_3对Jurkat细胞株hdpr1基因去甲基化作用机制的研究  

Mechanism of As_2O_3 on hdpr1 Gene Demethylation in Jurkat Cell Line

作　　者：沈松菲[1] 沈建箴[1] 付海英[1] 吴淡森[1] 徐成波[1] 朱艺芳[1] 

机构地区：[1]福建医科大学附属协和医院血液病研究所,福建福州350001

出　　处：《中国实验血液学杂志》2010年第6期1484-1488,共5页Journal of Experimental Hematology

基　　金：福建省自然基金(编号:CO540014);福建医科大学重大科研项目计划课题任务书(编号:09ZD021)

摘　　要：本研究探讨三氧化二砷(arsenic trioxide,As2O3)对急性T淋巴细胞白血病Jurkat细胞株hdpr1抑癌基因去甲基化的影响及其作用机制。采用CCK8法检测As2O3对Jurkat细胞的增殖抑制作用,流式细胞术检测As2O3作用前后细胞周期的变化,甲基化特异性PCR(MSP)检测As2O3对hdpr1基因甲基化模式的影响,用半定量RT-PCR检测As2O3对hdpr1基因、DNA甲基转移酶基因dnmt1、dnmt3a、dnmt3b mRNA表达水平的影响。结果表明:As2O3可明显抑制Jurkat细胞的增殖并呈时间-浓度依赖性;As2O3阻滞Jurkat细胞周期于G0/G1期(p<0.05),呈浓度依赖性;As2O3能够逆转Jurkat细胞株hdpr1基因的高甲基化并诱导其mRNA重新表达,同时下调dnmt1、dnmt3a、dnmt3b mRNA的表达水平,亦呈浓度依赖性。结论:As2O3抑制Jurkat细胞的增殖,阻滞细胞周期于G0/G1期,其机制可能为下调dnmt1、dnmt3a、dnmt3b基因的表达,诱导Jurkat细胞中异常甲基化的hdpr1基因去甲基化并使其恢复表达。This study was purposed to investigate the effect of As2O3 on the demethylation of anti-oncogene-hdpr1 gene of acute lymphoblastic leukemia cell line Jurkat in vitro and its mechanism.The inhibitory effect of As2O3 on the proliferation of Jurkat cells was assayed by CCK-8; the change of Jurkat cell cycle was detected by flow cytometry before and after using As2O3; the effect of As2O3 on the methylation model of hdpr1 gene was analyzed by methylation-specific PCR,and the effect of As2O3 on the expression of hdpr1 mRNA was analyzed by semiquantitative RT-PCR. The results showed that the proliferation rate of Jurkat cells was decreased significantly after being treated with As2O3,and in dose-and time-dependent manner; As2O3 blocked Jurkat cell cycle in G0/G1 phase in dose-dependent manner. As2O3 could reverse hypermethylation of hdpr1 gene and induce its mRNA reexpression,and down-regulate the dnmt1,dnmt3a,dnmt3b mRNA expression level also in dose-dependent manner. It is concluded that the As2O3 suppresses the proliferation of Jurkat cells and blocks cell cycle is G0/G1,its possible mechanism may be down-regulating mRNA expression level of dnmt1,dnmt3a and dnmt3b,induc demethylation of hdpr1 gene from abnormal hypermethylation status and activates its reexpression.

关 键 词：甲基化 hdpr1 三氧化二砷 急性T淋巴细胞白血病 JURKAT细胞 

分 类 号：R733.71[医药卫生—肿瘤] R979.1[医药卫生—临床医学]