耐百草枯人肺腺癌细胞株的建立及其生物学特性  

Establishment and characterization of a Paraquat-resistant human lung adenocarcinoma cell line A549/PQ

作　　者：洪广亮[1] 谭佳平 丁颖威[1] 赵光举[1] 李萌芳[1] 卢中秋[1] 

机构地区：[1]温州医科大学附属第一医院急诊科,浙江温州325015

出　　处：《温州医科大学学报》2015年第11期781-784,共4页Journal of Wenzhou Medical University

基　　金：浙江省医学创新学科建设计划(11-CX26);浙江省十二五高校重点学科;浙江省中医药重点学科建设计划(2012-XK-A28)

摘　　要：目的:建立耐百草枯(PQ)的人肺腺癌细胞株(A549/PQ),并初步研究其生物学特性。方法:采用浓度递增诱导法建立A549/PQ,光镜观察细胞形态变化,以乳酸脱氢酶(LDH)细胞毒性检测法、CCK-8法检测A549/PQ和亲本A549对PQ的敏感性,计算A549/PQ的半数抑制浓度(IC50)和耐药指数(RI),细胞计数法绘制细胞生长曲线和倍增时间,流式细胞技术分析细胞周期。结果:光镜下A549/PQ细胞变大、形态不规则、细胞间隙变大。PQ诱导A549/PQ细胞LDH释放明显低于其亲本细胞,差异有统计学意义(P<0.05)。PQ诱导A549/PQ细胞IC50为1716.33 μmol/L明显高于A549细胞的215.03μmol/L(P<0.05),RI为7.98。A549/PQ细胞倍增时间明显高于A549细胞,差异有统计学意义(P<0.05)。A549/PQ细胞G0/G1期细胞明显增多,S期细胞明显减少(P<0.05)。结论:成功构建对PQ耐药的A549细胞,其生物学特征与亲本细胞存在明显差异,为PQ毒理机制研究提供了良好细胞模型。Objective： To establish Paraquat （PQ）-resistant A549 cell line （A549/PQ） and explore its biological characteristics. Methods： The A549 cells were periodically co-cultured with PQ at gradually increasing con- centrations, and its biological characteristics were observed. The morphological features were checked with light microscope. Sensitivity to PQ was detected by LDH assay. The cell growth curve and doubling time were studied by cell counting. CCK-8 assay was used to measure the median inhibitory concentration （ICs0） and resistance index （RI） of A549/PQ cells and flow cytometry was applied to evaluate the cells cycle distribution. Results： Compared with A549 cells, the changes for larger size, irregular shape and much wide cell gap were found in A549/PQ cells. LDH release induced by PQ in A549/PQ cells was significantly lower than that of A549 cells （P〈0.05）. CCK-8 assay showed higher ICs0 in A549/PQ cells than that in A549 cells （1716.33 ＆ 215.03 pmol/L, P〈0.05） and RI was 7.98 in A549/PQ cells. Longer doubling time, increased G0/G1 phase cells and decreased cells in S phase were found in A549 cells （P〈0.05）. Conclusion： We successfully constructed a PQ-resistant A549 cell line with significantly different biological characteristics and therefore a good cellular model for PQ toxicity research.

关 键 词：百草枯 耐药株 生物学特性 

分 类 号：R459.7[医药卫生—急诊医学]