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

摘要

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.%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。