上皮-间质转化和胰岛素样生长因子Ⅰ型受体在非小细胞肺癌EGFR-TKIs获得性耐药中的作用  

The role of epithelial-mesenchymal transition and insulin-like growth factorⅠreceptor in acquired resistance to epidermal growth factor-tyrosine kinase inhibitors in non-small cell lung cancer

作　　者：王津京[1] 张为民[2] 陈蓓[2] 林建光[1] 

机构地区：[1]南方医科大学广州临床医学院,广东广州510515 [2]广州军区广州总医院肿瘤科,广东广州510010

出　　处：《肿瘤》2013年第2期103-110,共8页Tumor

基　　金：国家自然科学基金资助项目(编号:81172225)

摘　　要：目的:探讨上皮-间质转化(epithelial-mesenchymal transition,EMT)和胰岛素样生长因子Ⅰ型受体(insulin-like growth factorⅠreceptor,IGF-1R)在非小细胞肺癌(non-smallcelllungcancer,NSCLC)对表皮生长因子受体-酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitors,EGFR-TKIs)获得性耐药中的作用。方法:选用EGFR基因突变型和野生型的人肺腺癌细胞PC-9和H460,分别用吉非替尼和厄洛替尼将其诱导成耐药的PC-9/ZD和H460/ER细胞。MTT法检测各组细胞的增殖情况,划痕实验及Transwell侵袭实验检测细胞迁移和侵袭能力,蛋白质印迹法和RT-PCR法分别检测IGF-1R及EGFR信号通路分子及EMT相关分子的蛋白和mRNA表达。结果:PC-9/ZD和H460/ER为EGFR-TKI获得性耐药细胞,分别对吉非替尼和厄洛替尼的敏感性显著下降,差异有统计学意义(P<0.05)。与PC-9和H460细胞相比,耐药的PC-9/ZD和H460/ER细胞形态呈现间质细胞表型,细胞侵袭和迁移能力增加(P<0.05)。PC-9/ZD和H460/ER细胞中间质标志分子vimentin的mRNA和蛋白表达量均显著增加(P<0.05),PC-9/ZD细胞的E-cadherin表达量显著减少(P<0.05)。与PC-9和H460细胞相比,PC-9/ZD与H460/ER细胞的IGF-1R及其磷酸化蛋白表达量显著增加(P<0.05),AKT和ERK磷酸化水平明显上调(P<0.05)。PC-9/ZD细胞的EGFR磷酸化水平与PC-9细胞相比未见明显差异(P>0.05),而H460/ER细胞的EGFR磷酸化水平较H460细胞显著降低(P<0.05)。结论:EGFR突变型PC-9细胞和野生型H460细胞的获得性耐药细胞中均发生了EMT,并且IGF-1R信号蛋白表达增多,提示EMT及IGF-1R信号转导通路均可能在NSCLC的EGFR-TKIs获得性耐药中起重要作用。Objective： To investigate the role of EMT （epithelial-mesenchymal transition） and IGF- 1R （insulin-like growth factor Ⅰ receptor） in acquired resistance to EGFR-TKIs （epidermal growth factor receptor-tyrosine kinase inhibitors） in NSCLC （non-small cell lung cancer）. Methods： The EGFR mutant human lung adenocarcinoma PC-9 cell line and the EGFR wild-type H460 cell line were used to generate gefitinib-resistant PC-9 cells （named as PC-9/ZD cells） and erlotinib-resistant cells （named as H460/ER cells）, respectively. MTT assay was used to measure the cell proliferation of PC9, PC-9/ZD, H460 and H460/ER cells. Wound-healing assay and Transwell assay were used to determine the migration and invasion capabilities of the cells. The protein and mRNA expressions of E-cadherin, vimentin, EGFR, ERK （extracellular signal-regulated kinase）, AKT （protein kinase B） and IGF-1R were determined by Western blotting and RT-PCR （reverse transcription PCR）, respectively. Results： Both PC-9/ZD and H460/ER cellsacquired resistance to EGFR-TKIs which was to say that the sensitivities to gefitinib and erlotinib in PC-9/ ZD and H460/ER cells were significantly decreased, respectively （P 〈 0.05）. Compared with PC-9 and H460 cells, the PC-9/ZD and H460/ER cells displayed mesenchymal phenotypes, and their capabilities of invasion and migration were enhanced （P 〈 0.05）. The expression of mesenchymal cell marker vimentin was increased in both PC-9/ZD and H460/ER cells （P 〈 0.05）, and the expression of E-cadherin was decreased in PC-9/ZD cells （P 〈 0.05）. The expressions of IGF-1R and its phosphorylated form in both PC-9/ZD and H460/ER cells were significantly increased （P 〈 0.05） as compared with those in the PC-9 and H460 cells, accompanied by up-regulation of phosphorylation levels of AKT and ERK （P 〈 0.05）. No significant difference was found in phosphorylation level of EGFR between PC-9 and PC-9/ZD cells （P 〈 0.05）, while the phosphorylation level of EGFR w

关 键 词：癌 非小细胞肺 抗药性 肿瘤 蛋白激酶抑制剂 胰岛素样生长因子Ⅰ 上皮-间质转化 表皮生 长因子受体-酪氨酸激酶抑制剂 

分 类 号：R734.2[医药卫生—肿瘤]