人肺腺癌顺铂耐药细胞A549/CDDP对铂类化疗药的敏感性及耐药基因表达改变的研究  

Sensitivities to Platinum Drugs and the Expressions of Drug Resistance Genes in the Human Lung Adenocarcinoma CDDP-resistant Cell Sublines A549/CDDP

作　　者：李壮华[1] 贾筠[1] 陈镜塘[1] 徐瑞华[2,3] 

机构地区：[1]东莞市人民医院肿瘤内科,广东东莞523000 [2]华南肿瘤学国家重点实验室,广东广州510060 [3]中山大学肿瘤防治中心内科,广东广州510060

出　　处：《赣南医学院学报》2016年第1期38-41,共4页JOURNAL OF GANNAN MEDICAL UNIVERSITY

摘　　要：目的:探讨人肺腺癌细胞系A549/CDDP对铂类化疗药耐药的机制。方法:应用MTT法检测人肺腺癌细胞A549及其顺铂耐药细胞A549/CDDP对铂类化疗药(CDDP、CBD及L-OHP)的敏感性,应用RT-PCR法比较两细胞系多药耐药基因(包括MDR1,ABCG2,MRP1,LRP),DNA-polβ和GST-pi等在mRNA水平的表达差异,筛选出差异基因,用Western blotting法验证该基因在蛋白水平的表达差异。结果:MTT实验发现,与人肺腺癌细胞系A549相比,顺铂耐药细胞A549/CDDP对CDDP的耐药系数(RI)为7.877,而A549/CDDP细胞也对CBD及LOHP交叉耐药(RI分别为4.486及2.352);RT-PCR发现两细胞系的MDR1,ABCG2,MRP1,LRP及GST-pi等耐药基因在mRNA水平无明显差异,但A549/CDDP细胞中DNA-polβ在mRNA及蛋白表达水平均较A549细胞明显升高。结论:A549/CDDP细胞内DNA-polβ表达升高可能与铂类耐药有关。Objective： To explore the mechanism for platinum-resistance in the human lung adenocarcinoma CDDP-resistant cell sublines A549 / CDDP. Methods： In CDDP-sensitive cell lines A549 and the corresponding CDDP-resistant cell subline A549 / CDDP,platinum drugs＇ sensitivities were determined by MTT assay,then multidrug resistance（ MDR）-related genes（ such as MDR1,ABCG2,MRP1,LRP）,DNA-polβand GST-pi were analyzed by RT-PCR,and then the changes at mRNA level were confirmed by Western-blotting at protein-levels. Results： CDDP-resistant cell sublines A549 /CDDP which was 7. 877 folds more resistant than the parental cell line A549 showed cross resistance to CBD and L-OHP（ 4. 486 and 2. 352 folds respectively）. Between A549 and A549 /CDDP cells,there were no significant differences in expressions of MDR1,ABCG2,MRP1,LRP and GST-pi; however,DNA-polβexpression in A549 / CDDP was significantly higher than A549 cell at both mRNA and protein levels. Conclusion： The results indicates that DNA-polβmay be related to the platinum-derivates resistance in A549 / CDDP.

关 键 词：铂类化疗药 多药耐药 DNA-polβ GST-PI 

分 类 号：R734.2[医药卫生—肿瘤]