LRP15基因启动子区甲基化与其表达的关系

Analysis of the Methylation in the Promoter of LRP15 Gene and Its Expression

为了研究LRP15基因启动子区的甲基化状况及其与基因表达的关系,探讨LRP15基因的甲基化调控机 制,采用甲基化特异性聚合酶链反应(MS-PCR)检测LRP15基因启动子区的甲基化状况,应用甲基化抑制剂5-杂 氮-2'-脱氧胞嘧啶(CdR)及去乙酰化酶抑制刑曲古抑菌素(TSA)诱导CpG岛去甲基化及组蛋白乙酰化,观察启动 子区CpG岛甲基化与基因表达的关系。结果显示:白血病细胞系K562中LRP15基因启动子区呈高甲基化状态, 并抑制了该基因的表达。单独应用或联合TSA应用CdR均可去除该基因启动子区的高甲基化状态.并诱导该基 因的表达。这表明LRP15基因的表达受到启动子区甲基化的调控;该基因在K562细胞系中不表达与其启动子区 高甲基化及组蛋白去乙酰化密切相关。结论:甲基转移酶抑制剂与去乙酰化酶抑制剂在白血病治疗中可能具有一 定的价值。

To study the methylation in the promoter of LRP15 gene and its relationship with gene expression and to explore the possible mechanism of regulating LRP15 gene methylation, the methylation in the promoter of LRP15 gene in K562 cell line was detected by MS-PCR. Then K562 was exposed to 5-aza-2'-deoxycytidine (CdR) and trichostatin (TSA) , to determine whether the silencing of LRP15 gene by de novo methylation could be reversed. As a result, it was confirmed by MS-PCR that the promoter of LRP15 was hypermathylated in K562 cell line, and lost its transcription activity. After CdR, with or without TSA, the silencing of LRP15 gene by de novo methylation can be reversed. Observation demonstrated that the expression of LRP15 was controlled by methylation in its promoter in K562. It is suggested that methyltransferase inhibitor and deacetylase inhibitor may be effective agents in leukemia therapy.