摘要
目的 检测白血病细胞株HL-60的死亡相关蛋白激酶1（death-associated protein kinase 1,DAPKl）基因启动子甲基化状态及其在白血病发病机制中的应用.方法 分别采用重亚硫酸盐测序PCR（bisulfite sequencing PCR,BSP）和逆转录PCR（reverse transcription PCR,RT-PCR）检测正常健康儿童（对照组）外周血单核细胞（peripheral blood mononuelear cell,PBMC）和白血病细胞株HL-60（研究组）的DAPKl基因启动子甲基化状态和DAPK1基因mRNA表达.结果 正常健康儿童外周血单核细胞DAPKl基因启动子CpG岛CG位点呈无或低甲基化状态（甲基化率≤10%）,DAPK1基因mRNA表达正常.白血病细胞株HL-60的DAPK1基因启动子表现呈高甲基化状态（甲基化率为60%～90%）,显著高于正常健康儿童外周血单核细胞,DAPKl基因mRNA表达呈缺失状态.结论 DAPKl基因启动子在白血病HL一60细胞株呈高甲基化状态,与白血病的发病机制存在关联.
Objective To study the methylation status of death associated protein kinase 1 （DAPK1） gene＇s promoter and its role in the pathogenic mechanism of leukemia. Methods DNA and RNA were extracted from a healthy child＇s peripheral blood mononuclear cells （PBMC） and leukemia cell line HL 60 and were amplified with bisulfite sequencing PCR （BSP） and reverse transcription PCR （RT PCR）, respectively. Methylation status and mRNA expression of DAPK1 gene were compared. Results Promoter of DAPK1 gene in healthy child＇s peripheral blood mononuclear cells displayed unmethylated or low methylated status with normal mRNA expression （methylation rate ≤ 10 %）, while it was highly methylated in leukemia cell line HL-60 with no mRNA expression （methylation rate 60%- 90%）. Conclusion Promoter of DAPK1 gene in leukemia cell line HL-60 was hypermethylated. It may correlate with pathogenic mechanism of leukemia.
出处
《中华妇幼临床医学杂志（电子版）》
CAS
2011年第3期189-191,共3页
Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)
关键词
外周血单核细胞
DAPK1基因
启动子
甲基化
白血病
儿童
peripheral blood mononuclear ceils （PBMC）
death-associated protein kinase 1 （DAPK1）
promoter
methylation
leukemia
child
