砷剂诱导多发性骨髓瘤细胞socs-1基因去甲基化作用的研究  

Arsenic Trioxide Induces socs-1 Gene Demethylation in Myeloma Cell Lines

作　　者：王鸣明 朱琦 任志宏 邹丽芳 窦红菊 胡钧培 

机构地区：[1]上海第九人民医院血液科,上海血液研究所,上海200011

出　　处：《中国实验血液学杂志》2008年第5期1064-1068,共5页Journal of Experimental Hematology

基　　金：上海交通大学医学院院基金课题,编号05XJ21023

摘　　要：本研究探讨三氧化二砷(As2O3)对人多发性骨髓瘤细胞株U266、RPMI8226细胞内socs-1基因甲基化状态的影响。用MTT法观察As2O3对骨髓瘤细胞增殖情况的影响,采用甲基特异性PCR法检测As2O3作用前后骨髓瘤细胞株U266、RPMI8226socs-1基因的甲基化状态,以实时定量PCR技术定量检测细胞株给药前后socs-1基因mRNA的表达变化,流式细胞技术检测As2O3诱导的骨髓瘤细胞的凋亡情况。结果表明:人骨髓瘤细胞株U266、RPMI8226均存在不同程度的socs-1基因CpG岛甲基化,socs-1基因不表达的现象。As2O3作用72小时后socs-1基因甲基化程度明显减弱或消失,socs-1基因在mRNA水平上表达明显增强,与各野生型细胞株相比,差异具有显著性p<0.05),且细胞生长抑制明显,早期、晚期细胞凋亡比率明显升高,并呈现剂量依赖性。结论:As2O3可诱导socs-1基因甲基化状态的改变,使基因表达上调,恢复其活性,这为进一步阐明As2O3诱导骨髓瘤细胞凋亡的可能机制和As2O3治疗多发性骨髓瘤的可能机制提供新的思路和新的研究方向。The aim of this study was to explore the effect of arsenic trioxide （ As2O3 ） on the methylation status of socs-1 gene in multiple myeloma cell lines U266, RPMI8226. The cell viability was assayed by MTT method. The methylation status of socs-1 gene was detected by methylation specific PCR. The expression of socs-1 gene mRNA was determined with real-time PCR. The cell apoptosis was analyzed by flow cytometry. The results indicated that hypermethylation of CpG island of socs-1 gene was observed without expression of socs-1 in myeloma cell lines U266. RPMI8226. The expression of socs-1 gene mRNA in each myeloma cell line increased significantly after exposure to As2O3 for 72 hours as compared with the cell lines of wild type （p 〈 0.05 ）. And cell proliferation was significantly inhibited, both early apoptosis and later apoptosis ratios increased in dose-dependent manner. It is concluded that As2O3 may induce socs-1 demethylation and up-regulate the expression of the gene. This study provides a new thought and direction for exploring possible mechanism of cell apoptosis induced by As2O3 and multiple myeloma treatment by As2O3.

关 键 词：三氧化二砷 骨髓瘤 SOCS-1基因 基因甲基化 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤] R979.1[医药卫生—临床医学]