Institute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 10081, China.
Department of Veterinary Biomedical Sciences, and Toxicology Center, University of Saskatchewan, 52 Campus Dr, Saskatoon, SK S7N 5B4, Canada; Department of Integrative Biology and Center for Integrative Toxicology, Michigan State University, 784 Wilson Rd, East Lansing, MI 48824, USA; Department of Environmental Science, Baylor University, 97266 One Bear Place, Waco, TX 76798, USA.
Ecotoxicol Environ Saf. 2022 Sep 15;243:113982. doi: 10.1016/j.ecoenv.2022.113982. Epub 2022 Aug 17.
Fluorene-9-bisphenol (BHPF), which has been used as a substitute for bisphenol A (BPA) in consumer goods and industrial products, can be detected in environmental media and human urine. BHPF has been reported to have endocrine-disrupting effects, whereas deleterious effects on steroidogenesis in H295R cells and underlying mechanisms are still unclear. Here, we investigated effects of BHPF on steroidogenesis using human adrenocortical carcinoma cells (H295R). Cytotoxicity was initially assessed and half-maximal inhibitory concentration (IC) was determined based on proliferation of cells. Responses of four steroid hormones, aldosterone, cortisol, testosterone and 17β-estradiol (E), and ten critical genes, StAR, HMGR, CYP11A1, CYP11B1, CYP11B1, HSD3B2, CYP21, CYP17, 17β-HSD, and CYP19, involved in steroidogenesis after exposure to non-cytotoxic concentrations of BHPF were determined in the presence or absence of 100 μM dbcAMP. Adenylate cyclase (AC) activity, intracellular concentrations of cAMP, PKA activity and amounts of steroidogenic factor-1 (SF-1) gene and expressions of proteins were determined to elucidate underlying mechanisms of effects on steroidogenesis. BHPF was cytotoxic to H295R cells in a dose- and time-dependent manner. Effects on production of hormones results demonstrated that exposure to greater concentrations of BHPF inhibited productions of aldosterone, cortisol, testosterone and E by down-regulation of steroidogenic genes. Inhibition of AC activity, intercellular cAMP content and PKA activity after exposure to BHPF implied that the AC/cAMP/PKA signaling pathway was involved in BHPF-induced suppression of steroidogenesis in H295R cells. Additionally, BHPF inhibited steroidogenesis and expressions of steroidogenic genes via decreasing expression of SF-1 protein, both in basal and dbcAMP-induced treatment. These results contributed to understanding molecular mechanisms of BHPF-induced effects on steroidogenesis and advancing the comprehensive risk assessment of BPs.
芴-9-双酚（BHPF）已被用作消费品和工业产品中双酚 A（BPA）的替代品，可在环境介质和人尿中检测到。据报道，BHPF 具有内分泌干扰作用，但其对 H295R 细胞类固醇生成的有害影响及其潜在机制尚不清楚。在这里，我们使用人肾上腺皮质癌细胞（H295R）研究了 BHPF 对类固醇生成的影响。首先评估细胞增殖的细胞毒性和半最大抑制浓度（IC）。在不存在或存在 100μM dbcAMP 的情况下，确定暴露于非细胞毒性浓度的 BHPF 后，四种类固醇激素（醛固酮、皮质醇、睾丸激素和 17β-雌二醇（E））和十个关键基因（StAR、HMGR、CYP11A1、CYP11B1、CYP11B1、HSD3B2、CYP21、CYP17、17β-HSD 和 CYP19）的类固醇生成反应。测定腺苷酸环化酶（AC）活性、细胞内 cAMP 浓度、PKA 活性和类固醇生成因子-1（SF-1）基因的量以及蛋白质的表达，以阐明对类固醇生成的影响的潜在机制。BHPF 以剂量和时间依赖的方式对 H295R 细胞具有细胞毒性。对激素产生的影响结果表明，暴露于较高浓度的 BHPF 通过下调类固醇生成基因抑制醛固酮、皮质醇、睾丸激素和 E 的产生。暴露于 BHPF 后 AC 活性、细胞内 cAMP 含量和 PKA 活性的抑制表明 AC/cAMP/PKA 信号通路参与了 BHPF 抑制 H295R 细胞类固醇生成的作用。此外，BHPF 通过降低 SF-1 蛋白的表达抑制类固醇生成和类固醇生成基因的表达，无论是在基础和 dbcAMP 诱导的治疗中。这些结果有助于了解 BHPF 对类固醇生成的影响的分子机制，并推进 BP 的综合风险评估。
