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目的探讨5-杂氮-2’-脱氧胞苷(5-aza-2dC)对HL-60细胞Galectin-1基因表达的影响,并进一步分析药物处理后Galectin-1基因启动子区域甲基化状态的改变。方法用5μmol/L的5-aza-2dC处理HL-60细胞,采用Real-TimeRT-PCR检测Galectin-1基因在药物处理与未处理组中的mRNA表达水平,甲基化特异性PCR(methylation-specific PCR,MSP)检测Galectin-1基因启动子区域在药物处理与未处理HL-60细胞中的甲基化水平。结果Real-TimeRT-PCR结果显示Galectin-1在处理后的HL-60细胞中表达上调,MSP结果显示,HL-60细胞Galectin-1基因启动子区存在高甲基化,经5-aza-2dC处理后能够使Galectin-1基因去甲基化。使其甲基化水平下降。结论启动子区域高甲基化是Galectin-1基因低表达的原因之一,5-aza-2dC能够使Galectin-1基因去甲基化,逆转Galectin-1基因的表达,5-aza-2dC可能在抗急性髓系白血病中发挥重要作用。
Objective To investigate the effect of 5-aza-2’-deoxycytidine (5-aza-2dC) on Galectin-1 gene expression in HL-60 cells and further analyze the methylation status of Galectin-1 gene promoter Change. Methods HL-60 cells were treated with 5μmol / L 5-aza-2dC. Real-time RT-PCR was used to detect the mRNA expression of Galectin-1 in drug-treated and untreated groups. Methylation- specific PCR (MSP) was used to detect the methylation level of Galectin-1 promoter in drug-treated and untreated HL-60 cells. Results Real-Time RT-PCR results showed that Galectin-1 was up-regulated in HL-60 cells treated with 5-aza-2dC, and MSP showed hypermethylation in Galectin-1 promoter region of HL-60 cells Galectin-1 gene can be demethylated. Its methylation level decreased. Conclusions Promoter hypermethylation is one of the reasons of low expression of Galectin-1 gene. 5-aza-2dC can demethylate Galectin-1 gene and reverse Galectin-1 gene expression. 5-aza- Acute myeloid leukemia plays an important role.