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Effects of Epidermal Growth Factor/Hydrocortisone on the Growth and Differentiation of Human Ovarian Surface Epithelium

Sarah L. Maines-Bandiera, MSc

Peter C. K. Leung, PhD

Yu-Long Hu, PhD

Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada; Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California

Nelly Auersperg, MD, PhD

Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco, San Francisco, California; Department of Obstetrics and Gynecology, University of British Columbia, B.C. Women's Hospital, Rm. 2H30, 4490 Oak Street, Vancouver B.C., V6H 3V5, Canada auersper{at}interchange.ubc.ca

Objective: Ovarian surface epithelium (OSE), the precursor of the epithelial ovarian carcinomas, has limited growth potential in culture. Epidermal growth factor+hydrocortisone (EGF+HC) enhances its growth but induces epitheliomesenchymal transition (EMT). This study was undertaken to define the effects of EGF+HC and their reversibility, to optimize growth-promoting media, and to relate OSE phenotypes in vitro to physiologic states in vivo.

Methods: OSE was cultured in media 199/MDCB105 or EBM (Clonetics) with 2% or 10% fetal bovine serum with or without 10 ng/mL EGF, 1.0 µg/mL HC, and 1.0 µg/mL bovine brain extract. Growth rates and growth potentials (population doublings [PD] to senescence) were defined, and growth patterns and expression of keratin and collagen types III and IV were compared with the ovarian cancer cell lines OVCAR3 and SKOV3.

Results: EGF+HC increased growth potentials from 12-14 PD to 40-42 PD and reduced PD time from 53 hours to 20 hours. Without EGF+HC, OSE cells remained uniformly epithelial. EGF+HC induced EMT (mesenchymal shapes, reduced keratin, and production of collagenous extracellular matrix), but the EMT response varied greatly among OSE from different women. EMT was reversed over 1-2 weeks by seculture into EGF+HC-free medium in passage 1, but inconsistently thereafter. EGF+HC had no effect on the differentiation of ovarian carcinoma lines.

Conclusion: The phenotype of intact OSE in vivo is most closely reproduced in media without EGF+HC. EGF+HC enhances growth but initiates EMT, which likely mimics a repair response. Variations in EGF+HC-induced phenotypes point to the existence of OSE subpopulations with differing responsiveness to growth factors or steroids, which may relate to their susceptibility to malignant transformation.

Key Words: Ovarian surface epithelium • ovarian cancer • epidermal growth factor • hydrocortisone • epitheliomesenchymal transition • collagen secretion

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