【摘要】 本研究探讨白血病细胞系WT1基因的甲基化状况及其与表达的关系。用甲基化抑制剂和组蛋白脱乙酰基酶抑制剂成功诱导WT1表达。用地西他滨、曲古抑菌素处理白血病U937、HL-60、K562细胞系,用RT-PCR、改良的硫化PCR结合限制性内切酶技术检测mRNA表达。结果表明,U937细胞不表达WT1基因,而HL-60、K562和KG1细胞高表达WT1基因;HL-60细胞WT1基因没有甲基化,而K562和U937细胞发生了甲基化。地西他滨、曲古抑菌素单药处理U937细胞不能诱导WT1基因表达,而二药联合作用可以诱导U937细胞系WT1基因重新表达。结论:单纯DNA甲基化不能抑制白血病细胞WT1基因表达;DNA甲基化与组蛋白脱乙酰基共同抑制了WT1基因表达;地西他滨、曲古抑菌素联合作用可以重新诱导WT1基因表达。更多还原

【Abstract】 This study was aimed to investigate the methylation status of WT1 gene in leukemia cell lines and its relation with expression of WT1 gene.The WT1 gene was silenced by DNA methylation or histone deacetylation,and the expression of WT1 gene was induced by using HDAC inhibitor and/or demethylation agent of DNA.Some leukemia cell lines(U937,HL-60,K562,KG1) were detected by RT-PCR,MS-PCR,restriction analysis,and DNA sequencing.U937 leukemic cells without WT1 mRNA expression were incubated with HDAC inhibitor Trichostatin A(TSA) and/or demethylation agent decitabine.The results showed that the U937 cells did not express WT1 gene,but HL-60,K562 and KG1 cells hyghly expressed WT1 gene;WT1 gene was unmethylated in HL-60 cells,but methylated in K562 and U937 cells.WT1 expression could be reactivated by co-incubation with TSA and decitabine,but not was observed by using single drug.It is concluded that WT1 promoter is methylated in some leukemia cells,however,the methylation can not affect its expression.DNA methylation and deacetylation of histones are synergistic to inhibit the expression of WT1 in leukemic U937 cells,the combination of TSA with decitabine can induce expression of WT1 gene.更多还原