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目的检测白血病细胞雌激素受体(estrogen receptors,ER)基因启动子区CpG岛甲基化状态情况,探讨雌激素受体基因多个启动子区包括ERα(ERα-A,-Band-C)和ERβ甲基化与其表达异常的联系。方法应用RT-PCR检测6种白血病细胞株以5′-杂氮-2-脱氧胞苷(5′-Aza-Dc)去甲基化前后ER基因表达活性并应用甲基化特异性PCR技术(methylation specific PCR,MSP)检测10例正常人白细胞和6种白血病细胞株以5′-Aza-Dc去甲基化前后ER基因甲基化状态。结果ER各启动子在白血病细胞株中均出现甲基化或半甲基化带,但只有ERα-A基因表达沉默而且在正常人白细胞中呈未甲基化状态,在以5′-Aza-Dc处理细胞株后ERα-A基因表达恢复。结论白血病细胞的雌激素受体基因启动子区存在高甲基化现象并导致其基因表达沉默,可能为白血病的致病机制分析提供了一个新的方向,提示ERα-A基因启动子CPG岛的甲基化有可能成为白血病的新的参考标记物。
Objective To detect the methylation status of CpG island in estrogen receptor (ER) gene promoter region of leukemia cells and to explore the relationship between estrogen receptor (ERα-A, -Band-C) ERβ methylation and its abnormal expression. Methods RT-PCR was used to detect ER gene expression in 6 leukemia cell lines before and after demethylation with 5’-aza-2-deoxycytidine (5’-Aza-Dc). Methylation-specific PCR methylation specific PCR (MSP) was used to detect the methylation status of ER gene in 10 normal human leukemia cells and 6 leukemia cell lines before and after demethylation of 5’-Aza-Dc. Results All the ER promoters showed methylation or semi-methylation in leukemia cell lines. However, only the ERα-A gene was silenced and was unmethylated in normal human leukocytes. In the presence of 5’-Aza- ERα-A gene expression recovered after Dc treatment. Conclusion The hypermethylation of estrogen receptor gene promoter region in leukemia cells leads to silent gene expression, which may provide a new direction for the pathogenesis analysis of leukemia. It suggests that the methylation of ERα-A promoter on CPG island It may become a new reference marker for leukemia.