GnRH-(1-5) transactivates EGFR in Ishikawa human endometrial cells via an orphan G protein-coupled receptor

Madelaine Cho-Clark; Darwin O Larco; Nina N Semsarzadeh; Florencia Vasta; Shaila K Mani; T John Wu

doi:10.1210/me.2013-1203

GnRH-(1-5) transactivates EGFR in Ishikawa human endometrial cells via an orphan G protein-coupled receptor

Mol Endocrinol. 2014 Jan;28(1):80-98. doi: 10.1210/me.2013-1203. Epub 2013 Jan 1.

Authors

Madelaine Cho-Clark¹, Darwin O Larco, Nina N Semsarzadeh, Florencia Vasta, Shaila K Mani, T John Wu

Affiliation

¹ Department of Obstetrics and Gynecology (T.J.W., M.C., F.V.) and the Program in Molecular and Cellular Biology (D.O.L., T.J.W.), Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, Maryland 20814; and Departments of Molecular and Cellular Biology and Neuroscience (S.K.M.), Baylor College of Medicine, Houston, Texas 77030.

Abstract

The decapeptide GnRH is known for its central role in the regulation of the hypothalamo-pituitary-gonadal axis. In addition, it is also known to have local effects within peripheral tissues. The zinc metalloendopeptidase, EC 3.4.24.15 (EP24.15), can cleave GnRH at the Tyr(5)-Gly(6) bond to form the pentapeptide, GnRH-(1-5). The central and peripheral effect of GnRH-(1-5) is different from its parent peptide, GnRH. In the current study, we examined the effect of GnRH-(1-5) on epidermal growth factor receptor (EGFR) phosphorylation and cellular migration. Using the Ishikawa cell line as a model of endometrial cancer, we demonstrate that GnRH-(1-5) stimulates epidermal growth factor release, increases the phosphorylation of EGFR (P < .05) at three tyrosine sites (992, 1045, 1068), and promotes cellular migration. In addition, we also demonstrate that these actions of GnRH-(1-5) are mediated by the orphan G protein-coupled receptor 101 (GPR101). Down-regulation of GPR101 expression blocked the GnRH-(1-5)-mediated release of epidermal growth factor and the subsequent phosphorylation of EGFR and cellular migration. These results suggest that GPR101 is a critical requirement for GnRH-(1-5) transactivation of EGFR in Ishikawa cells.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Calcium Signaling
Cell Line, Tumor
Cell Movement
Endometrial Neoplasms
Epidermal Growth Factor / metabolism
ErbB Receptors / antagonists & inhibitors
ErbB Receptors / genetics*
ErbB Receptors / metabolism
Female
Gene Expression
Gonadotropin-Releasing Hormone / physiology*
Humans
Matrix Metalloproteinase Inhibitors / pharmacology
Oligopeptides / physiology
Oncogene Proteins / genetics
Oncogene Proteins / metabolism*
Peptide Fragments / physiology*
Phenylalanine / analogs & derivatives
Phenylalanine / pharmacology
Phosphorylation
Protein Processing, Post-Translational
Pyrrolidonecarboxylic Acid / analogs & derivatives
Quinazolines / pharmacology
Receptors, Cell Surface / genetics
Receptors, Cell Surface / metabolism*
Receptors, G-Protein-Coupled / genetics
Receptors, G-Protein-Coupled / metabolism*
Receptors, LHRH
Thiophenes / pharmacology
Transcriptional Activation*
Tyrphostins / pharmacology

Substances

ADGRF1 protein, human
GPAnt-2
GPR101 protein, human
LHRH (1-5)
Matrix Metalloproteinase Inhibitors
Oligopeptides
Oncogene Proteins
Peptide Fragments
Quinazolines
Receptors, Cell Surface
Receptors, G-Protein-Coupled
Receptors, LHRH
Thiophenes
Tyrphostins
RTKI cpd
Gonadotropin-Releasing Hormone
Phenylalanine
Epidermal Growth Factor
batimastat
EGFR protein, human
ErbB Receptors
Pyrrolidonecarboxylic Acid

Grants and funding

This work was supported by The National Science Foundation (IOB-0544150 and IOS-1052288) (to T.J.W.), Department of Defense Grant (RO85FN) (to T.J.W.), and Henry M. Jackson Foundation Graduate Fellowship (to D.O.L.).