Extracellular vesicles shed from gefitinib-resistant nonsmall cell lung cancer regulate the tumor microenvironment
Extracellular vesicles shed from gefitinib-resistant nonsmall cell lung cancer regulate the tumor microenvironment
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DOI：
10.1002/pmic.201400008
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发表时间：
2014-08-01
期刊：
影响因子：
3.4
通讯作者：
Kim, Jayoung
中科院分区：
文献类型：
--
作者：
Choi, Do-Young;You, Sungyong;Kim, Jayoung
表皮生长因子受体（EGFR）-酪氨酸激酶抑制剂（TKI），包括吉非替尼，是携带激活EGFR突变的非小细胞肺癌患者的一线治疗选择，然而，对EGFR-TKI的获得性耐药是不可避免的。本研究的主要目的是利用蛋白质组学分析鉴定来自吉非替尼耐药的非小细胞肺癌细胞的胞外囊泡（EV）的信息蛋白质特征。Nano-LC-MS/ MS分析以高置信度（错误发现率< 0.05，倍数变化>= 2）鉴定了在PC 9 R细胞中富集的664种EV蛋白，其由于EGFR T790 M突变而对吉非替尼耐药。计算分析表明，包括AKT（也称为PKB，蛋白激酶B）/mTOR（雷帕霉素的机械靶点）途径在内的几种信号转导机制的组分在来自PC 9 R细胞的EV中过度表达。用从PC 9 R细胞收获的EV处理受体细胞增加了信号分子的磷酸化，并增强了对吉非替尼诱导的凋亡的增殖、侵袭和耐药性。AKT/mTOR通路的剂量和时间依赖性药物抑制克服了PC 9 R细胞和具有EGFR T790 M突变的H1975细胞的耐药性。我们的研究结果为调节肿瘤微环境的致癌EV蛋白特征提供了新的见解，并将有助于开发预测和评估吉非替尼耐药性的新诊断策略。
Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs), including gefitinib, are the first-line treatment of choice for nonsmall cell lung cancer patients who harbor activating EGFR mutations, however, acquired resistance to EGFR-TKIs is inevitable. The main objective of this study was to identify informative protein signatures of extracellular vesicles (EV) derived from gefitinib-resistant nonsmall cell lung cancer cells using proteomics analysis. Nano-LC-MS/ MS analysis identified with high confidence (false discovery rate < 0.05, fold change >= 2) 664 EV proteins enriched in PC9R cells, which are resistant to gefitinib due to EGFR T790M mutation. Computational analyses suggested components of several signal transduction mechanisms including the AKT (also PKB, protein kinase B)/mTOR (mechanistic target of rapamycin) pathway are overrepresented in EV from PC9R cells. Treatment of recipient cells with EV harvested from PC9R cells increased phosphorylation of signaling molecules, and enhanced proliferation, invasion, and drug resistance to gefitinib-induced apoptosis. Dose-and time-dependent pharmaceutical inhibition of AKT/mTOR pathway overcame drug resistance of PC9R cells and those of H1975 exhibiting EGFR T790M mutation. Our findings provide new insight into an oncogenic EV protein signature regulating tumor microenvironment, and will aid in the development of novel diagnostic strategies for prediction and assessment of gefitinib resistance.