title: Modulation of steroidogenic gene expression and hormone synthesis in H295R cells exposed to PCP and TCP.
abstract: Chlorophenols (CPs) have been suspected to disrupt the endocrine system and thus affect human and wildlife reproduction but less is known about the underlying mechanism. In this study, we investigated the effects of pentachlorophenol (PCP) and 2,4,6-trichlorophenol (TCP) on human adrenocortical carcinoma cell line (H295R). The H295R cells were exposed to environmentally relevant concentration (0.0, 0.4, 1.1, 3.4μM) of PCP and TCP for 48h, and expression of specific genes involved in steroidogenesis, including cytochrome P450 (CYP11A, CYP17, CYP19), 3βHSD2, 17βHSD4 and StAR was quantitatively measured using real-time polymerase chain reaction. The selected gene expressions were significantly down-regulated compared with those in the control group. Exposure to PCP and TCP significantly decreased production of both testosterone (T) and 17β-estradiol (E2). Furthermore, a dose-dependent decrease of cellular cAMP was observed in H295R cells exposed to both PCP and TCP. A time-course study revealed that the observed selected steroidogenic gene expressions and protein abundance (StAR) are consistent with reduced cellular cAMP concentrations. The results showed that PCP and TCP may inhibit steroidogenesis by disrupting cAMP signaling. The research indicates that H295R cells can be used as an in vitro model for endocrine disruption assay for chlorophenols and the mechanism involvement of disturbing cAMP signaling.
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
authors.forename: Yanbo
authors.initials: Y
authors.lastname: Ma
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Liu
authors.forename: Paul K S
authors.initials: PK
authors.lastname: Lam
authors.forename: Rudolf S S
authors.initials: RS
authors.lastname: Wu
authors.forename: John P
authors.initials: JP
authors.lastname: Giesy
authors.forename: Markus
authors.initials: M
authors.lastname: Hecker
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhang
authors.forename: Bingsheng
authors.initials: B
authors.lastname: Zhou
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002733:Chlorophenols / Q000633:toxicity*; D000242:Cyclic AMP / Q000378:metabolism; D052244:Endocrine Disruptors / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000096:biosynthesis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D012739:Gonadal Steroid Hormones / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010416:Pentachlorophenol / Q000633:toxicity*; D010750:Phosphoproteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013739:Testosterone / Q000096:biosynthesis; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D002733:Chlorophenols; D052244:Endocrine Disruptors; D012739:Gonadal Steroid Hormones; D010750:Phosphoproteins; D000098682:Steroidogenic Acute Regulatory Protein; D013739:Testosterone; D004958:Estradiol; D010416:Pentachlorophenol; D000242:Cyclic AMP; C024564:2,4,6-trichlorophenol