摘要
目的:研究顺铂(DDP)对肺癌细胞FANCC、FANCF、FANCL表达水平的影响,探讨FA/BRCA途径DNA损伤修复功能在肺癌细胞对顺铂耐药机制中的作用。方法:采用CCK-8法测定经不同浓度顺铂处理24、48 h后两种肺癌细胞株(A549和Calu-1)的细胞增殖抑制率。应用实时荧光定量PCR技术(RFQ-PCR)和蛋白质印迹法检测两类肺癌细胞株中的FANCC、FANCF、FANCL mRNA和蛋白的表达。结果:肺癌A549细胞和Calu-1细胞的增殖抑制率均随顺铂浓度和作用时间的不同而变化,呈剂量依赖性(P<0.05)和时间依赖性(P<0.05),Calu-1细胞的半数抑制浓度(IC50)明显高于A549细胞。肺癌A549细胞除在10μg/ml浓度顺铂处理24 h后FANCF和FANCL mRNA表达水平增高外,2.5～5μg/ml和20μg/ml浓度顺铂时FANCF和FANCL mRNA呈无变化和低表达(P<0.05)。细胞Calu-1经不同浓度顺铂处理后的FANCC和FANCF mRNA表达水平先增高后降低(P<0.05)。肺癌A549细胞中的FANCC、FANCF和FANCL蛋白表达水平随顺铂浓度增高呈进行性降低。而肺癌Calu-1细胞的FANCF蛋白表达水平在顺铂浓度5～10μg/ml范围内较对照组明显增高。结论:Calu-1细胞对顺铂的耐药性显著高于A549细胞,其机制之一可能是FA/BRCA途径中FANCF蛋白高表达的结果。
Objective： To study the effect of cisplatin on expression levels of FANCC,FANCF,FANCL in lung cancers,and explore the effect of FA/BRCA pathway in the mechanism of cisplatin resistance of lung cancer cells.Methods： Inhibition rate of lung cancer cell lines（A549 and Calu-1）after treatment of variant concentration cisplatin was measured using the CCK-8 assay.The expression levels of FANCC,FANCF and FANCL mRNA was detected by real-time fluorogentic quantitative PCR（RFQ-PCR）.The expression of FANCC,FANCF,FANCL proteins was determined by Western blotting.Results： The rate of proliferation inhibiting of lung cancer cells is increased in line with the prolong time of cisplatin treatment（P〈0.05） and the increase of cisplatin concentration（P〈0.05）.The inhibitory concentration of 50 percent in Calu-1 cells is significantly higher than in A549 cells.After treatment of 5-10 μg/ml and 20μg/ml cisplatin,FANCF and FANCL mRNA expression levels in A549 cells are similar to control group and markedly lower compared with control group,respectively.However,FANCC and FANCF mRNA levels in Calu-1 cells are initiatory increased and then decreased after treatment of variant concentrations cisplatin.The expressions of FANCC,FANCF and FANCL protein in A549 cells were decreased during the treatment of 5-20 μg/ml cisplatin as compared with control group.In Calu-1 cells,the expression levels of FANCF protein were significantly increased compared with control group during the treatment of 5-10 μg/ml cisplatin.Conclusion： The cisplatin resistance in Calu-1 cells is significantly increased than in A549 cells.The higher expression of FANCF protein of FA/BRCA pathway may be one of mechanisms for increased resistant to cisplatin in Calu-1 cells.
出处
《江苏大学学报（医学版）》
CAS
2012年第3期236-241,共6页
Journal of Jiangsu University：Medicine Edition
关键词
肺癌细胞
顺铂
FA/BRCA途径
耐药
lung cancer cell
cisplatin
Fanconi anemia-BRCA pathway
drug resistance
