表皮生长因子受体突变细胞系H1650耐药机制探讨  

Molecular Mechanism of Erlotinib Resistance in Epidermal Growth Factor Receptor Mutant Non-small cell Lung Cancer Cell Line H1650

作　　者：韩瑞丽[1] 王小丽[1] 钟殿胜[2,3] 赵娟[1] 陈哲[1] 孙琳琳[3] 王竞[3] 张金棒[1] 

机构地区：[1]天津医科大学总医院呼吸科,天津300052 [2]肿瘤科,天津300052 [3]天津市肺癌研究所,天津300052

出　　处：《中国肺癌杂志》2012年第12期689-693,共5页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金资助项目(No.30971307,No.81071915);天津市应用基础及前沿技术研究(No.10JCYBJC13700)资助~~

摘　　要：背景与目的表皮生长因子受体(epidermal growth factor receptor,EGFR)高表达和突变与40%左右的肺癌有关,已成为靶向治疗药物研究热点;随着Gefitinib和Erlotinib作为EGFR酪氨酸激酶抑制剂(tyrosine kinaseinhibitor,TKI)代表药物应用于临床,继而产生的耐药现象亦成为临床一大难题,部分耐药机制仍不清楚。本研究探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞系H1650耐药机制。方法选用real-time RT-PCR检测EGFR野生型NSCLC细胞系中EGFR mRNA表达水平;MTT检测癌细胞对Erlotinib的药物敏感性;Western blot检测EGFR突变NSCLC细胞系突变情况和Erlotinib及PI3K抑制剂(LY294002)对EGFR突变型NSCLC细胞下游信号蛋白磷酸化水平的影响。结果 EGFR野生型细胞系中,EGFR mRNA表达水平高低不一,但均对Erlotinib耐药;EGFR突变型细胞系中,HCC827和H1650为同种突变类型,HCC827对Erlotinib敏感,H1650则相对耐药;检测显示,H1650细胞中PTEN表达缺失,给予Erlotinib和LY294002处理后,HCC827中p-AKT明显被抑制,但H1650中p-AKT下调不明显。结论在NSCLC细胞系中,Erlotinib药物敏感性与EGFR的mRNA表达高低无关,但与EGFR的突变类型有关;H1650对Erlotinib相对耐药可能与PTEN缺失导致的p-AKT持续活化有关。Background and objective Epidermal growth factor receptor （EGFR） overexpression and mutations were existed in more than 40% of the lung cancer, and it＇s the one of molecular targets in dinical treatment. But the EGFR tyro- sine kinase inhibitors （TKI）-resistance is becoming a challenging clinical problem as following the application of EGFR-TKIs, Gefitinib or Erlotinib. However, the mechanistic explanation for resistance in the some cases is still lacking. Here we researched the resistance mechanism of H1650 cells. Methods Using real-time RT-PCR to analyze the EGFR mRNA expression level in EGFR wild-type non-smaU cell lung cancer （NSCLC） cells; M＇IT analysis detected the cytotoxicity for NSCLC cells to Er- lotinib; Western blot analysis examined the mutant situations and the downstream signaling protein phosphorylation level in EGFR-mutant NSCLC cells with the treatment of Erlotinib or/and PI3Kinhibitor, LY294002. Results In the EGFRwild-type NSCLC cells, the expression level of EGFR mRNA varied dramatically and all the ceils showed resistant to Erlotinib; In the EGFR-mutant ceils, HCC827 and H1650 （the same activating-mutation type）, HCC827 cells were Erlotinib-sensitive as well as FI1650 demonstrated primary relative resistance. Western blot analysis showed the loss of PTEN and the p-AKT level was not inhibited with the treatment of Erlotinib or/and LY294002 in H1650 cells, while HCC827 cells were no PTEN loss and definitively decrease ofp-AKT level. Conclusion EGFR wild-type NSCLC ceUs were resistant to Erlotinib no matter of how EGFR mRNA expression level. EGFR-activating mutations correlated with responses to Erlotinib. The PTEN loss and activa-tion of AKT signaling pathway contributed to Erlotinib resistance in EGFR-mutant NSCLC cell line H1650.

关 键 词：肺肿瘤 表皮生长因子受体 突变 Erlotinib耐药 

分 类 号：R734.2[医药卫生—肿瘤]