Erlotinib-induced autophagy in epidermal growth factor receptor mutated non-small cell lung cancer
Erlotinib-induced autophagy in epidermal growth factor receptor mutated non-small cell lung cancer
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DOI：
10.1016/j.lungcan.2013.05.012
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发表时间：
2013-09-01
期刊：
影响因子：
5.3
通讯作者：
Ho, James Chung-man
中科院分区：
文献类型：
--
作者：
Li, Yuan-yuan;Lam, Sze-kwan;Ho, James Chung-man
目的：厄洛替尼是治疗非小细胞肺癌（NSCLC）的常用酪氨酸激酶抑制剂（TKI）。自噬是一种对压力和营养缺乏做出反应的分解代谢过程。方法：选取4株表皮生长因子受体（EGFR）突变状态不同（外显子19缺失、外显子19缺失、野生型和L 858 R/T790 M）的NSCLC细胞株（HCC 827、HCC 4006、H358和H1975），采用免疫组化方法检测细胞自噬对厄洛替尼的耐药情况。MTT法、结晶紫染色法和Annexin-V法检测细胞活力和凋亡。Western blot检测自噬蛋白。用吖啶橙子染色法测定酸性泡状细胞器（AVO）的形成。结果：与EGFR突变状态一致，HCC 827和HCC 4006细胞对厄洛替尼敏感，而H358和H1975细胞对厄洛替尼耐药。临床相关浓度的厄洛替尼治疗通过p53核转位、AMPK激活和mTOR抑制诱导敏感NSCLC细胞系中的自噬（增加LC 3 II表达、Atg-5/Atg 12结合、AVO形成和p62降解）。加入氯喹作为自噬抑制剂，增强了敏感细胞对厄洛替尼的敏感性。类似地，Atg 5或Beclin-1的沉默显著增加了两种敏感细胞系对厄洛替尼的敏感性。与此相反，有耐药H358和H1975细胞株厄洛替尼exposure.Conclusions后，没有诱导自噬：厄洛替尼可以诱导敏感的非小细胞肺癌细胞株与激活EGFR突变（外显子19 del）的凋亡和自噬。抑制自噬可以进一步增强EGFR突变NSCLC对厄洛替尼的敏感性，表明自噬可能是一种保护机制。(C)2013爱思唯尔爱尔兰有限公司版权所有。
Purpose: Erlotinib is a commonly used tyrosine kinase inhibitor (TKI) in non-small cell lung cancer (NSCLC). Autophagy is a catabolic process in response to stress and deprivation of nutrients. This study aims to investigate whether autophagy confers acquired resistance to erlotinib treatment in NSCLC.Methods: Four NSCLC cell lines (HCC827, HCC4006, H358 and H1975) with different epidermal growth factor receptor (EGFR) mutation status (exon 19 deletion, exon 19 deletion, wild-type and L858R/T790M respectively) were selected. MTT assay; crystal violet staining and Annexin-V assay were performed to determine cell viability and apoptosis. Autophagic proteins were detected by Western blot. Acidic vesicular organelle (AVO) formation was determined by acridine orange staining. Autophagy inhibitor (chloroquine) and RNA interference were used to demonstrate the biological effect of erlotinib-induced autophagy.Results: In line with EGFR mutation status, it was shown that both HCC827 and HCC4006 cells were sensitive to erlotinib, while H358 and H1975 cell lines were resistant. Erlotinib treatment at clinically relevant concentrations induced autophagy (increased LC3II expression, Atg-5/Atg12 conjugation, formation of AVO and p62 degradation) in sensitive NSCLC cell lines, via p53 nuclear translocation, AMPK activation and mTOR suppression. Addition of chloroquine, as an autophagy inhibitor, enhanced erlotinib sensitivity in sensitive cells. Similarly, silencing of Atg5 or Beclin-1 significantly increased sensitivity to erlotinib in both sensitive cell lines. In contrast, there was no induction of autophagy in resistant H358 and H1975 cell lines upon erlotinib exposure.Conclusions: Erlotinib can induce both apoptosis and autophagy in sensitive NSCLC cell lines with activating EGFR mutation (exon 19 del). Inhibition of autophagy can further enhance sensitivity to erlotinib in EGFR-mutated NSCLC, suggesting that autophagy may serve as a protective mechanism. (C) 2013 Elsevier Ireland Ltd. All rights reserved.