Molecular Mechanisms Mediating the Effect of Mono-(2-Ethylhexyl) Phthalate on Hormone-Stimulated Steroidogenesis in MA-10 Mouse Tumor Leydig Cells
Molecular Mechanisms Mediating the Effect of Mono-(2-Ethylhexyl) Phthalate on Hormone-Stimulated Steroidogenesis in MA-10 Mouse Tumor Leydig Cells
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DOI：
10.1210/en.2010-0010
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发表时间：
2010-07-01
期刊：
影响因子：
4.8
通讯作者：
Papadopoulos, Vassilios
中科院分区：
文献类型：
--
作者：
Fan, Jinjiang;Traore, Kassim;Papadopoulos, Vassilios
邻苯二甲酸二（2-乙基己基）酯是一种广泛使用的增塑剂，其活性代谢产物邻苯二甲酸单（2-乙基己基）酯（MEHP）已被证明对发育和成年动物的生殖道产生不良影响。然而，到目前为止，它们起作用的分子机制还不确定。在本研究中，我们使用系统生物学方法，探讨MEHP对MA-10 Leydig细胞基础和人绒毛膜促性腺激素（hCG）刺激的类固醇产生的影响的分子和细胞机制。MEHP诱导hCG刺激的类固醇形成呈剂量依赖性减少。通过基因微阵列和蛋白质高通量免疫印迹分析，分别测定了在hCG存在或不存在的情况下用浓度递增的MEHP处理的细胞中mRNA和蛋白质表达的变化。表达谱分析表明，低浓度的MEHP诱导了许多基因的表达，这些基因也在hCG刺激后表达。hCG和MEHP治疗之间的交叉比较揭示了两个基因，Anxa 1和AR 1。我们认为，这些基因可能参与了一个新的自我调节机制的类固醇。MEHP诱导的hCG刺激的类固醇形成的减少被活性氧产生的增加所抑制，后者由Cyp 1a 1基因及其网络介导。提出了MEHP作用于MA-10间质细胞类固醇生成的机制模型。（内分泌学151：3348-3362，2010）
Di-(2-ethylhexyl) phthalate, a widely used plasticizer, and its active metabolite, mono-(2-ethylhexyl) phthalate (MEHP), have been shown to exert adverse effects on the reproductive tract in developing and adult animals. As yet, however, the molecular mechanisms by which they act are uncertain. In the present study, we address the molecular and cellular mechanisms underlying the effects of MEHP on basal and human chorionic gonadotropin (hCG)-stimulated steroid production by MA-10 Leydig cells, using a systems biology approach. MEHP induced dose-dependent decreases in hCG-stimulated steroid formation. Changes in mRNA and protein expression in cells treated with increasing concentrations of MEHP in the presence or absence of hCG were measured by gene microarray and protein high-throughput immunoblotting analyses, respectively. Expression profiling indicated that low concentrations of MEHP induced the expression of a number of genes that also were expressed after hCG stimulation. Cross-comparisons between the hCG and MEHP treatments revealed two genes, Anxa1 and AR1. We suggest that these genes may be involved in a new self-regulatory mechanism of steroidogenesis. The MEHP-induced decreases in hCG-stimulated steroid formation were paralleled by increases in reactive oxygen species generation, with the latter mediated by the Cyp1a1 gene and its network. A model for the mechanism of MEHP action on MA-10 Leydig cell steroidogenesis is proposed. (Endocrinology 151: 3348-3362, 2010)