Effects of epidermal growth factor/hydrocortisone on the growth and differentiation of human ovarian surface epithelium

• Published in: Journal of the Society for Gynecologic Investigation Vol. 11; no. 4; p. 241
• Main Authors: Salamanca, Clara M, Maines-Bandiera, Sarah L, Leung, Peter C K, Hu, Yu-Long, Auersperg, Nelly
• Format: Journal Article
• Language: English
• Published: United States 01.05.2004
• Subjects: Adult; Cell Differentiation - drug effects; Cell Division - drug effects; Cells, Cultured; Collagen Type III - biosynthesis; Collagen Type III - genetics; Collagen Type IV - biosynthesis; Collagen Type IV - genetics; Epidermal Growth Factor - pharmacology; Epithelial Cells - cytology; Female; Fluorescent Antibody Technique; Gene Expression; Humans; Hydrocortisone - pharmacology; Keratins - biosynthesis; Keratins - genetics; Microscopy, Fluorescence; Ovary - cytology; Phenotype; Tumor Cells, Cultured
• ISSN: 1071-5576; 1556-7117
• DOI: 10.1016/j.jsgi.2003.10.010

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. 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 microg/mL HC, and 1.0 microg/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. 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 subculture into EGF+HC-free medium in passage 1, but inconsistently thereafter. EGF+HC had no effect on the differentiation of ovarian carcinoma lines. 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.