c-Met抑制剂SU11274在HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄罗替尼耐药的逆转作用  

c-Met inhibitor reverses HGF-induced resistance to Erlotinib in non-small lung cancer cells with different EGFR gene types

作　　者：吴美玉[1] 玄香兰[2] 王富佳[1] 安昌善[1] 

机构地区：[1]延边大学附属医院呼吸内科,吉林延吉133000 [2]延边第二人民医院呼吸内科,吉林延吉133000

出　　处：《中国肿瘤生物治疗杂志》2015年第4期427-431,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81160291)~~

摘　　要：目的:肝细胞生长因子(hepatocyte growth factor,HGF)诱导敏感非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞对表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth facter receptor tyrosine kinase inhibitor,EGFR-TKI)厄洛替尼耐药,本研究旨在探讨c-Met抑制剂SU11274逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞株对厄洛替尼耐药及逆转耐药机制。方法:选择人NSCLC细胞株PC9(EGFR突变型,敏感株)、H292(EGFR野生型,敏感株)和A549(EGFR野生型,原发性耐药株),应用厄洛替尼和SU11274(1μmol/L)单独或联合作用于HGF(40 ng/ml)诱导的细胞株,实验分为6组:C组(不加药对照组)、H组(HGF处理)、E组(厄洛替尼处理组)、S组(SU11274处理组)、HE组(HGF+厄洛替尼处理组)、HES组(HGF+厄洛替尼+SU11274处理组)。MTT法、流式细胞术分别检测不同药物处理对各细胞增殖、凋亡的影响;应用Western blotting检测不同药物处理对各细胞中c-Met及其下游通道Stat3、Akt、Erk1/2蛋白表达水平。结果:厄洛替尼对3种细胞的增殖抑制作用均呈浓度依赖性,HGF处理能够缓解厄洛替尼的增殖抑制作用(P<0.05);不同浓度厄洛替尼联合SU11274作用于HGF诱导细胞时3种细胞株存活率比厄洛替尼单独作用于HGF诱导细胞时明显降低(P<0.05);HGS组的细胞凋亡比HG组明显增加(P<0.05);HGS组的c-Met、Stat3、Akt、Erk1/2活化蛋白量比HG组明显减少。结论:c-Met抑制剂SU11274和厄洛替尼联合应用可逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄洛替尼耐药,其机制可能与抑制HGF活化的c-Met及其下游通道蛋白表达有关。Objective: To investigate whether c-Met inhibitor SU11274 can reverse resistance to Erlotinib,an epidermal growth factor receptor tyrosine kinase inhibitor( EGFR-TKI),induced by HGF in non-small cell lung cancer cells with different EGFR gene types. Methods: NSCLC cells with different EGFR gene types,including PC9( EGFR-activating mutant),H292( EGFR-wild type),and A549( EGFR-wild type) were utilized in the study. The experiments for each cell line consisted of six different treatment groups: C group( control),H group( HGF),E group( Erlotinib),S group( SU11274),EH group( Erlotinib + HGF),and ESH group( Erlotinib + SU11274 + HGF). Their effects on cell survival and apoptosis were measured by MTT assay and flow cytometry( FCM). The activation of c-Met,Stat3,Akt,and Erk1 /2 protein were examined by immunoblotting. Results: Erlotinib inhibited growth of the three cells lines in a dosedependent manner,and the inhibition was effectively blocked by HGF. The presence of SU11274 significantly decreased the survival rates of cells exposed to HGF and Erlotinib( P < 0. 05). Apoptosis in cells treated with Erlotinib,SU11274,and HGF was also markedly increased compared with these treated with Erlotinib and HGF only( P < 0. 05). Similarly,the levels of p-Met,p-Stat3,p-Akt,and p-Erk1 /2 in the HES group were significantly lower than that in the HE group( P < 0. 05). Conclusion: SU11274 reversed HGF-induced resistance to Erlotinib in non-small lung cancer cells with different EGFR gene type,likely due to the inhibition of HGF-induced activation of c-Met and its down streams signaling pathways.

关 键 词：肝细胞生长因子 厄洛替尼 耐药:SU11274 非小细胞肺癌 

分 类 号：R734.2[医药卫生—肿瘤]