K562和K562/DNR细胞中mdr1基因启动子甲基化状态的分析

Analysis of methylation status of the promoter of mdr1 gene in K562 and K562/DNR cells

目的 分析K5 6 2和K5 6 2 /DNR细胞中mdr1基因启动子甲基化模式及其与P gp表达的关系。方法 用流式细胞术和RT PCR方法检测两个细胞系中mdr1基因的表达 ,用亚硫酸氢钠脱氨基 DNA测序的方法分析mdr1基因启动子甲基化模式。结果 K5 6 2细胞不表达P gp ,mdr1基因启动子甲基化 ;K5 6 2 /DNR细胞P gp表达阳性 ,mdr1基因启动子非甲基化。 结论 K5 6 2和K5 6 2 /DNR细胞mdr1基因启动子甲基化模式不同 ,前者有甲基化修饰 ,后者无甲基化修饰 。

Objective To analyze the methylation patterns of mdr1 gene promotor in both K562 cell and K562/DNR cells and the relationship between promotor methylation and P gp expression. Methods mdr1 gene expression was analyzed by flow cytometry and RT PCR, methylation status of the promotor of mdr1 gene by bisulfite sequencing (including two GC box). Results mdr1 gene was found methylated at GC box and not expressed in K562 cells, but unmethylated and expressed respectively in K562/DNR cells. Conclusion The methylation patterns of mdr1 gene promotor in K562/DNR and K562 cells were different. mdr1 gene silencing was associated with the promotor methylation.