title: Di(2-ethylhexyl) phthalate inhibits antral follicle growth, induces atresia, and inhibits steroid hormone production in cultured mouse antral follicles.
abstract: Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental toxicant found in consumer products that causes ovarian toxicity. Antral follicles are the functional ovarian units and must undergo growth, survival from atresia, and proper regulation of steroidogenesis to ovulate and produce hormones. Previous studies have determined that DEHP inhibits antral follicle growth and decreases estradiol levels in vitro; however, the mechanism by which DEHP elicits these effects is unknown. The present study tested the hypothesis that DEHP directly alters regulators of the cell cycle, apoptosis, and steroidogenesis to inhibit antral follicle functionality. Antral follicles from adult CD-1 mice were cultured with vehicle control or DEHP (1-100 μg/ml) for 24-96 h to establish the temporal effects of DEHP on the follicle. Following 24-96 h of culture, antral follicles were subjected to gene expression analysis, and media were subjected to measurements of hormone levels. DEHP increased the mRNA levels of cyclin D2, cyclin dependent kinase 4, cyclin E1, cyclin A2, and cyclin B1 and decreased the levels of cyclin-dependent kinase inhibitor 1A prior to growth inhibition. Additionally, DEHP increased the mRNA levels of BCL2-associated agonist of cell death, BCL2-associated X protein, BCL2-related ovarian killer protein, B-cell leukemia/lymphoma 2, and Bcl2-like 10, leading to an increase in atresia. Further, DEHP decreased the levels of progesterone, androstenedione, and testosterone prior to the decrease in estradiol levels, with decreased mRNA levels of side-chain cleavage, 17α-hydroxylase-17,20-desmolase, 17β-hydroxysteroid dehydrogenase, and aromatase. Collectively, DEHP directly alters antral follicle functionality by inhibiting growth, inducing atresia, and inhibiting steroidogenesis.
authors.affiliation: Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. Electronic address: phannon2@illinois.edu.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Hannon
authors.affiliation: Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. Electronic address: kbran@illinois.edu.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Brannick
authors.affiliation: Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. Electronic address: Wei.Wang2@covance.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. Electronic address: drrupesh@yahoo.com.
authors.forename: Rupesh K
authors.initials: RK
authors.lastname: Gupta
authors.affiliation: Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802, USA. Electronic address: jflaws@illinois.edu.
authors.forename: Jodi A
authors.initials: JA
authors.lastname: Flaws
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D052244:Endocrine Disruptors / Q000633:toxicity*; D005260:Female; D005496:Follicular Atresia; D005786:Gene Expression Regulation; D012739:Gonadal Steroid Hormones / Q000378:metabolism*; D051379:Mice; D006080:Ovarian Follicle / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D012333:RNA, Messenger / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques
chemical_list: D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D017077:Culture Media, Conditioned; D052244:Endocrine Disruptors; D012739:Gonadal Steroid Hormones; D012333:RNA, Messenger; D004051:Diethylhexyl Phthalate; D013250:Steroid Hydroxylases
keywords: Antral follicle; Atresia; Di(2-ethylhexyl) phthalate; Ovary; Steroidogenesis