中文摘要：

目的研究肝细胞生长因子（HGF）诱导敏感非小细胞肺癌（NSCLC）细胞对厄洛替尼耐药的机制,观察c-Met及其下游信号通道蛋白是否参与HGF诱导不同基因型NSCLC细胞对厄洛替尼耐药。方法 2014年1月—2015年1月,选择人NSCLC细胞株PC-9〔表皮生长因子受体（EGFR）突变型,敏感株〕、H292（EGFR野生型,敏感株）及人胚肺成纤维细胞MRC-5细胞,通过ELISA法检测PC-9、H292、MRC-5细胞培养上清液中HGF水平。用MRC-5细胞培养上清液诱导PC-9、H292细胞,采用Western blotting法检测c-Met及其下游通道蛋白表达情况。将56只雌性、SPF级BALB/c裸鼠随机分为8组,每组7只。在PC-9细胞诱导模型中,对照组（C组）和厄洛替尼处理组（E组）裸鼠皮下接种PC-9细胞悬液,MRC-5诱导组（H组）、MRC-5和厄洛替尼处理组（HE组）裸鼠皮下接种PC-9＋MRC-5细胞悬液;当移植瘤直径达到4 mm时,C组和H组采用0.9%氯化钠溶液灌胃,E组和HE组采用厄洛替尼灌胃。在H292细胞诱导模型中,C组、E组裸鼠皮下接种H292细胞悬液,H组、HE组裸鼠皮下接种H292＋MRC-5细胞悬液;模型建立后灌胃方式同PC-9细胞诱导模型。给药结束后处死裸鼠,比较PC-9、H292细胞诱导模型中各组移植瘤重量。采用免疫组化法检测裸鼠移植瘤组织中c-Met及其下游通道蛋白表达水平。结果 PC-9、H292细胞培养上清液中均未检测到HGF,MRC-5细胞培养上清液中HGF水平为（1 262±90）pg/ml。Western blotting法结果显示,MRC-5细胞培养上清液中HGF能活化PC-9、H292细胞中p-Met、p-Akt、p-Stat3、磷酸化细胞外调节蛋白激酶1/2（p-Erk1/2）活性。PC-9细胞诱导模型中：E组移植瘤重量小于C组（P〈0.05）;HE组移植瘤重量小于H组,大于E组（P〈0.05）。H292细胞诱导模型中：E组移植瘤重量小于C组（P〈0.05）;HE组移植瘤重量小于H组,大于E组（P〈0.05）。c-Met、p-Met分别定位于细胞膜和细胞质。在PC-9、H292细胞诱?

英文摘要：

Objective To study the mechanism of erlotinib resistance induced by hepatocyte growth factor（ HGF） in non- small- cell lung cancer（ NSCLC） cells and to investigate whether c- Met and its downstream signaling pathway proteins participate in the HGF- induced erlotinib resistance of NSCLC cells with different genetypes in vivo. Methods This study was conducted from January 2014 to January 2015. We selected NSCLC cell lines with different EGFR genes（ PC-9： EGFR-mutation type,sensitive; H292： EGFR- wild type,sensitive） and human embryonic lung fibroblasts MRC-5. The HGF level in cell culture supernatants secreted by PC-9,H292 and MRC-5 cells was quantified by ELISA. PC-9 and H292 cells were induced by MRC-5 cell culture supernatant; the expressions of c- Met and its downstream signaling pathway proteins were examined by Western blotting. Fifty- six female and SPF BALB / c nude mice were randomly divided into 8 groups,with each group containing 7. In building the PC-9 cell-induced model,the control group（ group C） and erlotinib treatment group（ group E） were subcutaneously inoculated PC-9 cell suspension and MRC-5 induced group（ group H）,MRC-5 and erlotinib treatment group（ group HE） were inoculated PC-9 ＋ MRC-5 cell suspension subcutaneously. In establishing the H292cell-induced model,group C and group E were inoculated H292 cell suspension subcutaneously while group H and group HE were inoculated H292 ＋ MRC-5 cell suspension subcutaneously. When the tumor diameter reached 4 mm in all groups,group C and group H were lavaged with 0.9% sodium chloride solution while group E and group HE were lavaged with erlotinib,respectively,in both built models. Mice were euthanized at after dosing. Comparisons of the weight of transplanted tumor were made between all groups in models induced by PC- 9 and H292 cells. The expressions of c- Met and its downstream signaling pathway proteins in transplanted tumor tissues in nude mice were determined via immunohistochemistry. Results The concentration o