雷公藤内酯醇对急性T淋巴细胞白血病Jurkat细胞系APC基因去甲基化诱导表达机制的研究  

Experimental study on triptolide reversing hypermethylation of APC in acute lymphoblastic leukemia cell line Jurkat

作　　者：吴雪梅[1] 沈建箴[1] 喻爱芳[1] 范丽萍[1] 付海英[1] 沈松菲[1] 吴淡森[1] 

机构地区：[1]福建医科大学附属协和医院血液病研究所,福州350001

出　　处：《白血病．淋巴瘤》2009年第6期327-330,共4页Journal of Leukemia & Lymphoma

基　　金：基金项目：福建省自然科学基金（C0540014）;福建省百千万人才工程基金（303052801）

摘　　要：目的以抑癌基因即腺瘤性结肠息肉病相关基因（APC）存在异常甲基化的急性T淋巴细胞白血病Jurkat细胞为研究对象，探究雷公藤内酯醇（TPL）对APC基因的影响，并对其机制进行初步探讨。方法运用四甲基偶氮唑盐（MTF）法等检测TPL对Jurkat细胞株增生的影响。巢式甲基特异性PCR（n—MSP）检测TPL对Jurkat细胞APC基因甲基化模式的影响。半定量RT—PCR检测经TPL作用后Jurkat细胞APC基因、甲基转移酶DNMT3A、DNMT3BmRNA的表达。Western blotting检测TPL作用后APC蛋白的表达。结果TPL对Jurkat细胞生长有明显的抑制作用，并有时间及剂量依赖性，48h IC50韧为19．7ng／ml；TPL可逆转APC基因的高甲基化；TPL能够诱导Jurkat细胞APC基因mRNA的表达，具剂量依赖性；TPL能够诱导Jurkat细胞APC蛋白重新表达，亦有剂量依赖性。结论小剂量TPL可明显抑制Jurkat细胞的增生，其可能机制为通过诱导Jurkat细胞中异常甲基化的APC基因去甲基化，使APC基因恢复表达。Objective To observe the effects of triptolide（TPL） on the anti-oncogene--APC gene of acute lymphoblastic leukemia cell line Jurkat in vitro. Methods The effects of TPL on proliferation Jurkat cells were assayed by using cell culture, MTT, The effects of TPL on APC gene of Jurkat cells were analyzed by nested methylation specific PCR and RT-PCR. The effects of TPL on the proteinum expression of APC gene were detected by Western blotting analysis. Results Following the treatment of TPL, the cell proliferation rate was degraded as the treatment concentration increased and the culture time extended. The effects were close and time-dependent. The 48 hour IC50 was 19.7 ng/ml. TPL can reverse hypermethylation of APC gene,and induce the expression of the mRNA and the proteinum. Conclusion Low dose TPL could depress the proliferation rate of Jurkat. The possible mechanism might be its reversing the hypermethylation of APC gene and activiting the expression of APC gene.

关 键 词：甲基化 基因 APC 雷公藤内酯醇 白血病 T细胞 急性 JURKAT细胞 

分 类 号：R733[医药卫生—肿瘤]