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Detection of 5-Aza-2'-deoxycytidine induced p16 gene demethylation in the multiple myeloma cell line U266
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- Author:
- No author available
- Journal Title:
- JOURNAL OF LEUKEMIA & LYMPHOMA
- Issue:
- 2
- DOI:
- 10.3760/cma.j.issn.1009-9921.2007.02.001
- Key Word:
- 多发性骨髓瘤;细胞系;基因表达调控;基因,肿瘤,抑制
Abstract： 目的 探讨5-氮-2'-脱氧胞嘧啶(5-Aza-CdR)诱导骨髓瘤细胞系U266p16基因DNA5'CpG岛去甲基化作用及对U266细胞增生的影响.方法 采用巢式甲基特异性PCR法(n-MSP)、DNA序列分析、RT-PCR、细胞生长曲线、流式细胞仪DNA含量分析法检测5-Aza-CdR对U266细胞p16基因去甲基化作用及其对U266细胞的生长、增生及细胞周期的影响.结果 (1)5-Aza-CdR能够逆转U266细胞p16基因异常甲基化;(2)5-Aza-CdR能激活p16基因沉默的再转录;(3)5-Aza-CdR能下调U266细胞甲基转移酶DNMT1、DNMT3A、DNMT3B的表达并呈浓度依赖性;(4)5-Aza-CdR作用的U266细胞被阻滞于G0～G1期.结论 5-Aza-CdR可能通过抑制甲基转移酶直接对p16基因去甲基化,逆转U266细胞DNA异常甲基化,并有效地激活因高甲基化所致p16基因沉默的再转录.
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