Visualization and quantitation of epidermal growth factor receptor homodimerization and activation with a proximity ligation assay
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Keiichi Ota1, Taishi Harada1,2, Kohei Otsubo1, Akiko Fujii1, Yuko Tsuchiya1, Kentaro Tanaka1, Isamu Okamoto1 and Yoichi Nakanishi1,3
1Research Institute for Diseases of The Chest, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan
2Department of Respiratory Medicine, Japan Community Health Care Organization (JCHO) Kyushu Hospital, Kitakyushu City, Fukuoka, 806-8501, Japan
3Center for Clinical and Translational Research, Kyushu University Hospital, Higashi-ku, Fukuoka 812-8582, Japan
Taishi Harada, email: email@example.com
Keywords: epidermal growth factor receptor (EGFR), lung cancer, proximity ligation assay, receptor dimerization, tyrosine kinase inhibitor (TKI)
Received: February 06, 2017 Accepted: June 27, 2017 Published: July 25, 2017
Objectives: Activation of the epidermal growth factor receptor (EGFR) results from receptor homodimerization and autophosphorylation and confers sensitivity to tyrosine kinase inhibitors in some tumors. However, the visual detection and quantitation of activated EGFR in the clinical setting has not been established.
Materials and Methods: A proximity ligation assay (PLA) was applied to detect EGFR homodimers in non–small cell lung cancer (NSCLC) cell lines and tissue specimens.
Results: PLA signals corresponding to EGFR homodimers were higher in NSCLC cell lines and tissue specimens positive for activating EGFR mutations than in those wild type (WT) for EGFR. Stimulation with EGF in NSCLC cells WT for EGFR or forced overexpression of EGFR in Ba/F3 cells resulted in marked EGFR homodimerization. The extent of EGFR homodimerization appeared related to that of EGFR autophosphorylation in NSCLC cells WT for EGFR.
Conclusion: PLA may provide a new tool for detection and quantitation of EGFR homodimers in NSCLC and other tumors.
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