5-Aza-CdR对CEM/C1白血病细胞KLF4基因的去甲基化作用

Effect of KLF4 gene of acute lymphoblastic leukemia cells treated with demethylated drug 5-Aza-CdR

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 approach the effect of promoter methylation to KLF4 gene of acute lymphoblastic leukemia cell line CEM/Cl and 5 - Aza - CdR impact on promoter methylation above. Methods Using different concentrations of 5 - Aza - CdR treatment of acute lymphoblastic leukemia cell line CEM/Cl, by thiazolyl blue （MTT） to detect the methylation drug 5 - Aza - CdR on CEM/C1 cell proliferation effect, and semi quantitative reverse transcriptase polymerase reaction （ RT - PCR） was used to detect the expression of KLF4 mRNA in CEM/Cl cells after the demethylating drugs 5 - Aza - CdR treatment. Results After 5 - Aza - CdR treatment, the proliferation of acute lymphoblastic leukemia cell line CEM/Cl is inhibited. Conclusion 5 -Aza- CdR can reactivate the KLF4 gene transcription and mRNA expression by demethylation, slow the growth of acute lymphoblastic leukemia cell line.