人胚胎骨髓MSC及其突变株F6细胞DAPK基因表达及启动子甲基化检测

摘要：目的：研究人胚胎骨髓间充质干细胞（fetal mesenchymal stem cells, FMSC）及其突变肿瘤细胞F6死亡相关蛋白激酶（deathassociated protein kinase,DAPK）基因的表达及启动子区域甲基化。 方法：用RT-PCR和甲基化特异性PCR（methylationspecific PCR, MS-PCR）技术对FMSC及F6细胞系DAPK 基因的表达和启动子甲基化状态进行检测，并对甲基化产物测序鉴定。 结果：DAPK基因在FMSC中表达而在F6中不表达。通过MS-PCR检测证实F6中DAPK基因发生甲基化，FMSC中DAPK基因发生未完全甲基化。 结论：DAPK基因启动子区域发生甲基化可能是促使其基因表达下调的重要机制，在FMSC突变成F6肿瘤细胞过程中发挥重要作用。

Detection of methylation of DAPK gene promoter in human bone marrow mesenchymal stem cells and its transformed mutant F6 cells

Objective To determine expression levels of death-associated protein kinase(DAPK) gene and analyze the methylation status of DAPK promoter in human fetal mesenchymal stem cells(FMSC) and its mutant F6 tumor cells.Methods The expression of DAPK gene was detected by RT-PCR.The methylation of DAPK promoter was detected by methylation specific PCR(MS-PCR).The products of methylation were confirmed by sequencing.Results DAPK was expressed in FMSC,but not in F6 cells.The promoter of DAPK gene was methylated in F6...