中国全科医学2017，Vol.20Issue(14)：1707-1711,5.DOI:10.3969/j.issn.1007-9572.2017.14.012
SU11274逆转肝细胞生长因子诱导的非小细胞肺癌细胞对厄洛替尼耐药作用及其机制研究
Mechanism of SU11274 Reversing the Resistance to Erlotinib Induced by Hepatocyte Growth Factor in Non-small Cell Lung Cancer Cells
摘要
Abstract
Background Hepatocyte growth factor(HGF) can induce non-small cell lung cancer(NSCLC) cells exerting resistance to epidermal growth factor receptor-tyrosine kinase inhibitors(EGFR-TKIs)in vitro,suggesting that HGF induced c-Met phosphorylation may be associated with the mechanism of erlotinib resistance in sensitive NSCLC.Objective The study aimed to demonstrate that c-Met inhibitor SU11274 can reverse the erlotinib resistance induced by HGF in sensitive NSCLC cells in nude mice.Methods This study was conducted from March 2014 to January 2015.Two cell lines including H292 and human embryonic fibroblast MRC-5 were chosen.The HGF produced by MRC-5 cells in cell culture supernatants was quantitated by enzyme-linked immunosorbent assay(ELISA).The proliferation of H292 cells was measured by using MTT assay and the expressions of c-Met and its downstream signaling proteins were examined by Western blotting.We selected forty-nine 6-week old nude mice and equally divided them into 7 groups,group C (control group),group H,group HS,group HD,group E,group HE,group HES,treated with H292 cell suspension,HGF,HGF+SU11274,HGF+1% DMSO,erlotinib,HGF+erlotinib,HGF+erlotinib+SU11274,respectively.The tumor growth was observed and the growth curve was drawn.The tumor weight was recorded.The expressions of c-Met and downstream signaling proteins in tumor tissues were determined via immunohistochemistry.Results ELISA assay found that the HGF level in the culture supernatant of MRC-5 cells was (1 262±90) pg/ml.MTT assay showed that in the mixture of H292 and MRC-5 cells culture supernatant with erlotinib concentration of IC50,the proliferation of H292 cells was 97.07%.The expressions of p-Met,p-STAT3,p-AKT and p-ERK1/2 in the culture supernatant of H292 with MRC-5 cells were higher than those in the H292 cell culture supernatant(P<0.05).Intervention method and duration exerted significant interaction effects on the tumor volume(P<0.001).Intervention method produced main effect on the tumor volume(P<0.001).Intervention duration had main effect on the tumor volume(P<0.001).The tumor volume was smaller in HES group than in HE group measured on the 7th,11th,14th,18th,21st and 25th days of intervention (P<0.05).The tumor mass differed significantly between the groups(F=39.120,P<0.001).HES group had less tumor mass than HE group did(P<0.05).Significant differences in the expressions of c-Met,p-Met,p-STAT3,p-AKT and p-ERK1/2 were found between the groups(P<0.05).The expressions of c-Met,p-Met,and p-STAT3 proteins were lower in HES group than those in HE group(P<0.05).Conclusion HGF secreted by MRC-5 induces erlotinib resistance in H292 cells in nude mice.c-Met inhibitor SU11274 reverses the resistance of HGF-induced sensitive NSCLC cells to erlotinib in vivo.The mechanism may be related to the inhibition of HGF-induced activation of c-Met and its signaling pathway proteins.关键词
癌,非小细胞肺/肝细胞生长因子/原癌基因蛋白质c-Met/SU11274/厄洛替尼Key words
Carcinoma,non-small-cell lung/Hepatocyte growth factor/Proto-oncogene proteins c-Met/SU11274/Erlotinib分类
医药卫生引用本文复制引用
严春花,吴叶丹,安昌善..SU11274逆转肝细胞生长因子诱导的非小细胞肺癌细胞对厄洛替尼耐药作用及其机制研究[J].中国全科医学,2017,20(14):1707-1711,5.基金项目
国家自然科学基金资助项目(81160291) （81160291）
