β-catenin对非小细胞肺癌细胞A549顺铂耐药作用及机制的探讨  

Role and mechanism of β-catenin-mediated resistance to cisplatin in human non-small cell lung cancer A549 cells

作　　者：钱海云[1,2] 黄江平[2] 崔丰和[2] 黄杰[1] 

机构地区：[1]武汉大学人民医院胸外科,湖北武汉430060 [2]荆州市中心医院胸心外科,湖北荆州434020

出　　处：《中华肿瘤防治杂志》2012年第21期1620-1624,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨β-catenin介导的信号通路在人非小细胞肺癌顺铂(DDP)耐受中的作用及可能机制。方法:MTS法检测人非小细胞肺癌A549及其衍生的DDP耐受细胞A549/DDP对DDP的敏感性;蛋白质印迹法检测A549和A549/DDP细胞内β-catenin及其靶基因Survivin和Bcl-2的蛋白表达,并以TCF/LEF报告基因检测β-catenin的活性;实时定量PCR检测Survivin和Bcl-2的转录水平;将β-catenin反义寡核苷酸片段转染入A549/DDP细胞沉默β-catenin的表达,MTS检测干扰前后细胞对DDP的敏感性,实时定量PCR及蛋白质印迹法检测相关基因的表达。结果:A549/DDP对DDP的耐受性是A549细胞的3.14倍,P=0.006 9;A549/DDP细胞中β-catenin蛋白含量上调且其活性上调4.28倍(P=0.003 6),β-catenin的靶基因Survivin及Bcl-2在mRNA及蛋白水平均上调;干扰下调A549/DDP细胞中β-catenin明显下调了Survivin及Bcl-2的mRNA及蛋白水平,增加了对DDP的敏感性。结论:β-catenin在耐药细胞A549/DDP对DDP的耐受中起着重要作用,是逆转肺癌DDP耐受的可能潜在靶标。OBJECTIVE：To investigate the role and its possible mechanisms of β-catenin signaling pathway in the cis platin （DDP） resistance in human non small cell lung cancer cells. METHODS： MTS assay was used to detect the cytotoxie activity of DDP in A549 and A549/DDP cells. The protein expressions of β-eatenin, Survivin and Bcl-2 in A549 and A549/DDP cells were analyzed by western blot. β-catenin transcriptional activity was measured using/3 catenin-Tcf reporter gene assay. The transcriptional levels of Survivin and Bcl-2 were detected by Real time PCR. β-catenin was Knockdowned by specific antisense oligonucleotide （AS（）） in A549/DDP cells and subsequent chemosensitivity to DDP was detected by MTS assay. RESULTS：Compared to the A549 cells,the resistance index （RI） of A549/DI）P cells to DDP was 3. 14 （P= 0. 006 9）. The expression of β-catenin was increased and its activity also increased about 4. 28 folds （P= 0. 003 6） in A549/DDP ceils. So as its downstream targets of β- catenin,Survivin and Bcl-2 at both mRNA and protein level were upregulated. Knockdown of β- catenin significantly downregulated Survivin and gel 2 expression and enhanced the chemosensitivity of A549/DDP cells to DDP. CONCLUSION： The β-catenin pathway plays an important role in the che moresistance of A549 /DDP cells to DDP,as a potential target to prevent DDP resistance.

关 键 词：癌 非小细胞肺 顺铂 抗药性 肿瘤 

分 类 号：R734.2[医药卫生—肿瘤]