人肺腺癌多药耐药细胞系A549/cDDP的建立及相关基因表达检测  

Establishment of a Multidrug Resistance Cell Line A549/cDDP of Human Lung Adenocarcinoma and Expression Analysis of Multidrug Resistance-Associated Genes

作　　者：潘永成[1] 赵伟刚[1] 王建军[1] 刘全[1] 杨光海[1] 李劲松[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胸外科,武汉430030

出　　处：《中国肺癌杂志》2009年第3期187-191,共5页Chinese Journal of Lung Cancer

基　　金：湖北省科技攻关项目(No.2007AA301B32-2)资助~~

摘　　要：背景与目的现有研究表明,肺癌细胞多药耐药造成的化疗失败是影响患者生存率的主要原因。本研究旨在建立人肺癌多药耐药细胞系,研究其耐药特性和机制。方法应用人肺腺癌细胞株A549,采用顺铂(cDDP)大剂量间歇诱导法,建立多药耐药细胞系A549/cDDP。相差显微镜观察细胞形态和结构差异,绘制两种细胞的体外生长曲线。用MTT法检测该耐药细胞株对多种化疗药物的多药耐药性,采用流式细胞术检测该耐药细胞表面多耐药基因(MDR)的表达产物P糖蛋白(P-gp)、多药耐药相关蛋白(MRP)及谷胱甘肽硫转移系统(GSH/GST)的表达情况,应用RT-PCR检测MDR、MRP基因表达水平。结果A549/cDDP对多种抗癌药物耐药,各种抗癌药物IC_(50)显著提高:(70.5±4.9)%的A549/DDP细胞表面P-gp表达阳性,而对照细胞A549表达仅为(42.4±5.6)%;(29.4±2.9)%的A549/DDP细胞表面MRP表达阳性,而对照细胞A549表达仅为(21.4±3.5)%,二者有差异(t=11.94; t=5.56,P均<0.01);A549/cDDP和A549细胞的GSH/GST表达无明显变化(P>0.05)。RT-PCR检测发现A549/cDDP中的MDR mRNA及MRP mRNA高表达。结论建立的A549/cDDP具有明确的多药耐药性,其多药耐药基因MDR mRNA及MRPmRN4表达升高。Background and objective It has been proven that chemotherapy failure caused by muhidrug resistance in lung tumor cells is the main cause for the patient＇s survival rate. The aim of this study is to establish a muhidrug resistance cell line of human lung adenoeareinoma and study the mechanism of muhidrug resistance. Methods Human lung adenocarcinoma cell line A549 was induced to muhidrug resistance cell line A549/cDDP by intermittent administration of high dose of cisplatin （cDDP）. The multidrug resistance was detected by using MTT assay. The levels of expression of MDR-1 gene-coded P-glyeoportein （P-gp）, muhidrug resistance-associated protein （MRP）, and GSH/GST were examined by flow cytometric assay. The levels of expression of MDR and MRP gene were also detected by RT- PCR in both A549/cDDP and A549 cell lines. Results A549/cDDP was resistant to many anti-tumor agents. The IC50 of A549/cDDP was 16.87 times higher than that of A549. The expressions of P-gp and MRP in A549/cDDP were increased significantly to （70.5±4.9）% and （29.4±2.9）%, respectively, vs （42.4±5.6）% and （21.4±3.5）% in A549. There was no difference of the GSH/GST expression between A549/cDDPand A549 cells. Conclusion A549/cDDP is a model with muhidrug resistance and the levels of MDR and MRP mRNA expressions are remarkably higher in A549/cDDP than those in A549.

关 键 词：肺肿瘤 多药耐药 基因表达 

分 类 号：R734.2[医药卫生—肿瘤]