Ezrin Enhances EGFR Signaling and Modulates Erlotinib Sensitivity in Non-Small Cell Lung Cancer Cells.

Abstract

Ezrin is a scaffolding protein that is involved in oncogenesis by linking cytoskeletal and membrane proteins. Ezrin interacts with epidermal growth factor receptor (EGFR) in the cell membrane, but little is known about the effects of this interaction on EGFR signaling pathway. In this study, we established the biological and functional significance of ezrin-EGFR interaction in non-small cell lung cancer (NSCLC) cells. Endogenous ezrin and EGRF interaction was confirmed by co-immunoprecipitation and immunofluorescent staining. When expression of ezrin was inhibited, EGFR activity and phosphorylation levels of downstream signaling pathway proteins ERK and STAT3 were decreased. Cell fractionation experiments revealed that nuclear EGFR was significantly diminished in ezrin-knockdown cells. Consequently, mRNA levels of EGFR target genes AURKA, COX-2, cyclin D1, and iNOS were decreased in ezrin-depleted cells. A small molecule inhibitor of ezrin, NSC305787, reduced EGF-induced phosphorylation of EGFR and downstream target proteins, EGFR nuclear translocation, and mRNA levels of nuclear EGFR target genes similar to ezrin suppression. NSC305787 showed synergism with erlotinib in wild-type EGFR-expressing NSCLC cells, whereas no synergy was observed in EGFR-null cells. Phosphorylation of ezrin on Y146 was found as an enhancer of ezrin-EGFR interaction and required for increased proliferation, colony formation, and drug resistance to erlotinib. These findings suggest that ezrin-EGFR interaction augments oncogenic functions of EGFR and that targeting ezrin may provide a potential novel approach to overcome erlotinib resistance in NSCLC cells.

• Authors: Yasemin Saygideğer-Kont, Tsion Zewdu Minas, Hayden Jones, Sarah Hour, Haydar Çelik, Idil Temel, Jenny Han, Nese Atabey, Hayriye Verda Erkizan, Jeffrey A Toretsky, Aykut Üren
• Journal: Neoplasia (New York, N.Y.) (Neoplasia) Vol. 18 Issue 2 Pg. 111-20 (Feb 2016) ISSN: 1476-5586 [Electronic] United States
• PMID: 26936397 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Cytoskeletal Proteins
• Neoplasm Proteins
• Protein Kinase Inhibitors
• STAT3 Transcription Factor
• ezrin
• Erlotinib Hydrochloride
• EGFR protein, human
• ErbB Receptors

Topics

• Apoptosis (drug effects)
• Carcinoma, Non-Small-Cell Lung (drug therapy, genetics, pathology)
• Cell Line, Tumor
• Cytoskeletal Proteins (genetics, metabolism)
• Drug Resistance, Neoplasm (genetics)
• ErbB Receptors (genetics, metabolism)
• Erlotinib Hydrochloride (administration & dosage)
• Humans
• Mutation
• Neoplasm Proteins (biosynthesis)
• Phosphorylation
• Protein Binding (drug effects)
• Protein Kinase Inhibitors (administration & dosage)
• STAT3 Transcription Factor (biosynthesis)
• Signal Transduction (drug effects)