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目的:观察甲基化转移酶抑制剂5-杂氮-2’-脱氧胞苷(5-aza-2’-deoxyc ityd ine,5-aza-2dC)对人急性髓系白血病细胞系HL-60细胞分化及对膜联蛋白A1/A2(Annexin A1/A2)表达和甲基化状态的影响。方法:瑞氏染色和流式细胞术检测5-aza-2dC对HL-60细胞分化的影响;RT-PCR法检测药物处理HL-60细胞前后Annexin A1和A2基因mRNA的表达水平;甲基化特异性PCR(m ethylation-spec ific PCR,MSP)检测药物处理HL-60细胞前后An-nexin A1和A2基因启动子区域CpG岛的甲基化水平。结果:5-aza-2dC处理后HL-60细胞的髓系分化抗原CD11b的表达增强,细胞向成熟分化,且在0.5μmol/L时其促分化作用最明显;Annexin A1和A2基因在HL-60细胞中低表达,0.5μmol/L 5-aza-2dC处理HL-60细胞72 h后,Annexin A1和A2基因mRNA表达水平明显上调,而其启动子区域CpG岛甲基化水平明显降低。结论:5-aza-2dC具有促进白血病细胞分化的作用,Annexin A1和A2基因启动子去甲基化可能与5-aza-2dC诱导白血病细胞分化有关。
AIM: To investigate the effect of 5-aza-2’-deoxycitydine (5-aza-2dC) on human acute myeloid leukemia cell line HL-60 Cell Differentiation and Its Influence on Annexin A1 / A2 Expression and Methylation Status. Methods: Wright’s staining and flow cytometry were used to detect the effect of 5-aza-2dC on the differentiation of HL-60 cells. The mRNA expression of Annexin A1 and A2 was detected by RT-PCR before and after treatment with HL-60 cells. The methylation levels of CpG island in the promoter region of An-nexin A1 and A2 gene in drug-treated HL-60 cells were detected by m ethylation-specific PCR (MSP). Results: After treatment with 5-aza-2dC, the expression of myeloid differentiation antigen CD11b in HL-60 cells was enhanced and the cells differentiated to mature stage. The differentiation of HL-60 cells was most obvious at 0.5μmol / L. 60 cells, the expression of Annexin A1 and A2 gene mRNA was up-regulated in HL-60 cells treated with 0.5μmol / L 5-aza-2dC for 72 h, while the methylation of CpG island in promoter region was significantly decreased. Conclusions: 5-aza-2dC can promote the differentiation of leukemia cells. Demethylation of Annexin A1 and A2 promoters may be related to the differentiation of leukemic cells induced by 5-aza-2dC.