title: Extracellular vesicles shed from gefitinib-resistant nonsmall cell lung cancer regulate the tumor microenvironment.
abstract: 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.
authors.affiliation: Department of Applied Chemistry, College of Applied Science, Kyung Hee University, Yongin, Republic of Korea.
authors.forename: Do-Young
authors.initials: DY
authors.lastname: Choi
authors.forename: Sungyong
authors.initials: S
authors.lastname: You
authors.forename: Jae Hun
authors.initials: JH
authors.lastname: Jung
authors.forename: Jae Cheol
authors.initials: JC
authors.lastname: Lee
authors.forename: Jin Kyung
authors.initials: JK
authors.lastname: Rho
authors.forename: Kye Young
authors.initials: KY
authors.lastname: Lee
authors.forename: Michael R
authors.initials: MR
authors.lastname: Freeman
authors.forename: Kwang Pyo
authors.initials: KP
authors.lastname: Kim
authors.forename: Jayoung
authors.initials: J
authors.lastname: Kim
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics; D005109:Extracellular Matrix / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D000077156:Gefitinib; D006801:Humans; D008168:Lung / Q000187:drug effects / Q000473:pathology; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D017354:Point Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D040901:Proteomics; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011799:Quinazolines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D059016:Tumor Microenvironment / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D066246:ErbB Receptors; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases; D000077156:Gefitinib
keywords: AKT; Cell biology; Extracellular vesicles; Nonsmall cell lung cancer; mTOR signaling pathway