E-钙黏蛋白在人非小细胞肺癌细胞多西他赛耐药中的作用

目的:探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)A549、H1299、H358、H441多西他赛(docetaxel,DTX)耐药细胞株与上皮间质转化(epithelial-mesenchymal transition,EMT)的关系,并初步研究逆转E-cadherin的表达对NSCLC细胞多西他赛耐药性的影响。方法:应用Real-time PCR和Western blot方法检测上皮间质转化相关标志物E-cadherin、Vimentin、N-cadherin、Snail在亲本细胞和多西他赛耐药细胞之间的表达差异;应用慢病毒载体介导构建稳定表达E-cadherin的NSCLC多西他赛耐药细胞;MTS法检测NSCLC细胞多西他赛耐药特性。结果:与A549、H1299、H358、H441四株亲本细胞相比,多西他赛耐药细胞(A549DTX,H1299DTX,H358DTX,H441DTX)形态呈长梭形,呈上皮间质转化(EMT)样改变。多西他赛耐药细胞中E-cadherin表达下调,Vimentin、N-cadherin、Snail表达上调。成功构建了过表达E-cadherin的NSCLC多西他赛耐药细胞。过表达E-cadherin细胞株细胞形态与空载对照细胞株以及亲本耐药细胞株相比呈间质上皮转化(MET)样改变。MTS结果显示四种不同E-cadherin过表达的NSCLC多西他赛耐药细胞对多西他赛的敏感性均强于其亲本耐药细胞和空载对照细胞。结论:NSCLC多西他赛耐药细胞发生EMT样改变,上调E-cadherin能增加NSCLC多西他赛耐药细胞对多西他赛的敏感性。

E-cadherin involves in the development of docetaxel resistance in human non-small cell lung cancer cells

Objective:To investigate the involvement of epithelia-mesenchymal transition (EMT) in four non-small cell lung cancer (NSCLC) cells (A549,H1299,H358,H441) docetaxel (DTX) resistance,in addition,to determine the effects of reversal of E-cadherin expression on NSCLC docetaxel resistance.Methods:The mRNA or protein expression of EMT-related markers such as E-cadherin,Vimentin,N-cadherin and Snail in parental cells and docetaxel-resistant cells were detected by Real-time PCR or Western blot.Overexpression of E-cadherin was mediated by lentivirus.Cell viability was detected by MTS assay.Results:Compared with the four parental cells of A549,H1299,H358 and H441,all docetaxel-resistant cells (A549DTX,H1299DTX,H358DTX,H441DTX) showed fibroblast-like morphology.The expression of E-cadherin was down-regulated in all docetaxel-resistant cells,however,the expression of Vimentin,N-cadherin and Snail were up-regulated.Overexpressing E-cadherin in NSCLC docetaxel-resistant cells showed mesenchymal-epithelial transition (MET)-like changes compared with the empty vector group and the parental drug-resistant cells.Moreover,MTS results showed that all the E-cadherin overexpressing NSCLC docetaxel-resistant cells were more sensitive to docetaxel than their parental resistant cells and control cells.Conclusion:NSCLC docetaxel-resistant cells occur EMT-like changes.Overexpression of E-cadherin can increase the sensitivity of NSCLC docetaxel-resistant cells to docetaxel.