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Obstetrics and Gynecology International
Volume 2011, Article ID 896896, 8 pages
http://dx.doi.org/10.1155/2011/896896
Research Article

Blocking Epidermal Growth Factor Receptor Signaling in HTR-8/SVneo First Trimester Trophoblast Cells Results in Dephosphorylation of PKB /AKT and Induces Apoptosis

1The Reproductive Molecular Biology Laboratory, Division of Maternal-Fetal Medicine, Departments of Obstetrics and Gynecology and Biochemistry and Molecular Biology, The University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA
2Clinic of Fetal Medicine and Maternal Medicine and Obstetrics and Gynecology, 263 Farmington Avenue Farmington, CT 06030, USA
3Department of Biochemistry and Microbiology, Faculty of Pharmacy, University of Kalamoon, P.O. Box: 222, Deirattiah, Syria
4Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, MSC 105580, 2211 Lomas Boulevard. NE, Albuquerque, NM 87131, USA
5The Center of Reproductive Medicine and Infertility at the Weill Cornell University, CRMI 1305 York Avenue, New York, NY 10021, USA
6Department of Obstetrics and Gynecology, The University of Iowa, Hospitals and Clinics, 200 Hawkins Drive, Iowa City, IA 52242, USA

Received 31 March 2011; Accepted 3 June 2011

Academic Editor: Edmund F. Funai

Copyright © 2011 J. Bolnick et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

We identified a major peptide signaling target of EGF/EGFR pathway and explored the consequences of blocking or activating this pathway in the first trimester extravillous trophoblast cells, HTR-8/SVneo. A global analysis of protein phosphorylation was undertaken using novel technology (Kinexus Kinetworks) that utilizes SDS-polyacrylamide minigel electrophoresis and multi-lane immunoblotting to permit specific and semiquantitative detection of multiple phosphoproteins. Forty-seven protein phosphorylation sites were queried, and the results reported based on relative phosphorylation at each site. EGF- and Iressa-(gefitinib, ZD1839, an inhibitor of EGFR) treated HTR-8/SVneo cells were subjected to immunoblotting and flow cytometry to confirm the phosphoprotein screen and to assess the effects of EGF versus Iressa on cell cycle and apoptosis. EGFR mediates the phosphorylation of important signaling proteins, including PKBα/AKT. This pathway is likely to be central to EGFR-mediated trophoblast survival. Furthermore, EGF treatment induces proliferation and inhibits apoptosis, while Iressa induces apoptosis.