5-氮-2'-脱氧胞苷对肺癌A549细胞凋亡及范可尼贫血互补基团F表达的影响

Influence of 5-Aza-CdR on the apoptosis of lung cancer A549 cell and the expression of FANCF gene

目的探讨5-氮-2'-脱氧胞苷(5-Aza-CdR)对人肺癌A549细胞凋亡及抑癌基因范可尼贫血互补基团F(FANCF)基因表达的影响。方法以浓度为0.5、5、50μmol/L的5-Aza-CdR处理人肺癌细胞株A549,常规培养采用四唑盐(MTT)比色法观察细胞的生长活性,甲基化特异性聚合酶链反应(PCR)检测FANCF基因甲基化状态;以荧光定量PCR检测FANCF mRNA的表达,并用流式细胞仪检测细胞凋亡率。结果 5-Aza-CdR能明显抑制肿瘤细胞的生长,细胞增殖抑制率(CPIR)随5-Aza-CdR浓度和作用时间的不同而变化,呈剂量依赖性(P<0.005)和时间依赖性(P<0.001);药物处理后FANCF mRNA表达明显升高,细胞凋亡率与5-Aza-CdR剂量呈正相关(r=0.998,P<0.05)。结论 5-Aza-CdR能使FANCF基因去甲基化,促进细胞凋亡,增强抑癌功能,但同时有增加顺铂耐药的风险。

Objective To investigate the influence of 5-Aza-2′-deoxycytidine（5-Aza-CdR） on the apoptosis of lung cancer A549 cell and the expression of Fanconi anemia complementation group F（FANCF） gene.Methods A549 cells were treated with 5-Aza-CdR（0.5, 5 and 50 μmol/L, respectively）.The growth of A549 cells was observed by 3-（4, 5-dimethylthiazol）-3,5-diphenyltetrazolium bromide（MTT） assay.The methylation status of FANCF gene was observed by methylation specific polymerase chain reaction （ PCR）.The expression of FANCF mRNA was observed by fluorescence quantitation PCR.The apoptosis rate of A549 cells was analyzed by flow cytometry.Results A549 cells treated with 5-Aza-CdR displayed a slow growth.The rate of cell proliferation inhibiting （CPIR） for A549 cells changed with the concentration and treatment time of 5-Aza-CdR （P 〈0.005, P 〈0.001）.FANCF mRNA expression increased after treatmert.The apoptosis rates after treatment had a positive correlation with 5-Aza-CdR dose （r =0.998, P 〈0.05）.Conclusions 5-Aza-CdR can induce the apoptosis of A549 cells by inducing demethylation and thereby enchancing FANCF gene, enhancing tumor suppressor function, but it can increase the risk of resistance to cisplatin.