摘要
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.
出处
《检验医学》
CAS
2015年第5期507-511,共5页
Laboratory Medicine
基金
湖北省十堰市科技局资助项目(ZD2012015)
湖北医药学院优秀中青年科技创新项目(2011CXG02)
