Effects of Endocrine Disrupters on Corneal Epithelial Cell Line

Abstract

Abstract: : Purpose: The eye is always exposed to physical and chemical stimulations. Because of this, xenobiotics and antioxdative enzymes are expressed in corneal epithelial cells. In this study, we estimated profiles of gene expression of human corneal epithelial cell line (CEPI) stimulated by endocrine disrupters. Methods: CEPI was grown in keratinocyte growth medium 2 (KGM2), which was a serum-free medium with the addition of bovine pituitary extract, to approximately 70% confluence in flasks. The cells were stimulated by 10 µM endocrine disrupters, i.e. di-2-ethylhexyl phthalate (DEHP), mono-2-ethylhexyl phthalate (MEHP), flutamide (FL), bisphenol A (BPA), and estradiol (E2), and incubated for 72 hours. After incubation, the cells were washed by PBS and lysed to extract total RNA. The total RNA isolated by Trizol was used for estimation of gene expression by GeneChip microarray. Results: Profile of gene expression was changed by addition of endocrine disrupters. The varied number of gene expressions stimulated by FL was the highest at 107 and by MEHP is was the lowest 11. Coefficients of correlation of stimulated gene expression between each endocrine disrupter were calculated. The coefficient of correlation for DEHP vs FL is 0.84, BPA vs E2 is 0.74, and FL and E2 vs 0.75. Conclusions: This study suggests that GeneChip was able to analyze influence of endocrine disrupters for gene expression in CEPI, comprehensively, and influence of endocrine disrupter for gene expression was able to classify by difference of profile of gene expression.

Keywords: cornea: epithelium • gene/expression