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目的探讨启动子甲基化对CEM/C1白血病细胞KLF4基因的作用及5-Aza-CdR干预对启动子甲基化的影响。方法使用不同浓度的5-Aza-CdR处理急性淋巴细胞白血病CEM/C1细胞,采用噻唑蓝(MTT)检测去甲基化药物5-Aza-CdRa对CEM/C1细胞增殖的影响,并利用半定量逆转录聚合反应(RT-PCR)检测去甲基化药物处理后CEM/C1细胞中KLF4基因mRNA的表达水平。结果 5-Aza-CdR抑制体外培养的人急性淋巴细胞白血病CEM/C1细胞增殖,呈浓度依赖性。经5-Aza-CdR处理CEM/C1细胞后,RT-PCR检测KLF4 mRNA恢复了表达。结论启动子区高甲基化是急性淋巴细胞白血病CEM/C1细胞中KLF4基因失活的主要原因,5-Aza-CdR能逆转KLF4基因甲基化状态,从而调控KLF4基因表达并抑制细胞生长。
Objective To investigate the effect of promoter methylation on KLF4 gene in CEM / C1 leukemia cells and the effect of 5-Aza-CdR intervention on promoter methylation. Methods The acute lymphoblastic leukemia CEM / C1 cells were treated with different concentrations of 5-Aza-CdR. The effect of 5-Aza-CdRa demethylation drug on the proliferation of CEM / C1 cells was detected by MTT assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of KLF4 in CEM / C1 cells after demethylation treatment. Results 5-Aza-CdR inhibited the proliferation of human acute lymphoblastic leukemia CEM / C1 cells in a concentration-dependent manner. After CEM / C1 cells were treated with 5-Aza-CdR, the expression of KLF4 mRNA was restored by RT-PCR. Conclusions Hypermethylation of promoter region is the main reason of KLF4 gene inactivation in acute lymphoblastic leukemia CEM / C1 cells. 5-Aza-CdR can reverse KLF4 methylation status and regulate KLF4 gene expression and inhibit cell growth.