Oskarsson Agneta, Ullerås Erik, Plant Kathryn E, Hinson Joy P, Goldfarb Peter S
School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK.
J Appl Toxicol. 2006 Nov-Dec;26(6):484-92. doi: 10.1002/jat.1166.
The focus on the refinement, reduction and replacement of animal use in toxicity testing requires the development of cell-based systems that mimic the effects of xenobiotics in human tissues. The human adrenocortical carcinoma cell line, H295R, has been proposed as a model for studies on adrenal steroidogenesis and its disruption. In this study, expression profiles for nine adrenal steroidogenic genes were characterized in H295R cells using real-time RT-PCR. Treatment with forskolin increased cortisol secretion and stimulated transcription of all the steroidogenic genes except SULT2A1. The transcript profile from H295R cells in the presence and absence of forskolin was compared with the transcript profile from human adrenal glands. The gene expression pattern observed in the forskolin-treated H295R cells was more similar to that in the human adrenal gland, than the expression pattern in untreated cells. To examine H295R cells as a possible in vitro system for the assessment of adrenal disruption using molecular endpoints, the insecticide lindane (gamma-hexachlorocyclohexane) was used. In vivo, lindane has been shown to inhibit testicular, ovarian and adrenal steroidogenesis. It was demonstrated that lindane reduced cortisol secretion, downregulated the expression of a subset of the genes encoding steroidogenic enzymes and repressed transcriptional activation of the steroidogenic acute regulatory protein (StAR) gene promoter. Thus the H295R cell line provides a good in vitro system for the analysis of the human adrenal steroidogenic pathway at the level of hormone production and gene expression. This in vitro test can be used for the rapid detection of adrenal endocrine disruption and as a tool for mechanistic studies.
在毒性测试中，对优化、减少和替代动物使用的关注要求开发能够模拟异生物素在人体组织中作用的细胞系统。人肾上腺皮质癌细胞系H295R已被提议作为研究肾上腺类固醇生成及其破坏作用的模型。在本研究中，使用实时逆转录聚合酶链反应（RT-PCR）对H295R细胞中9个肾上腺类固醇生成基因的表达谱进行了表征。用福司可林处理可增加皮质醇分泌，并刺激除SULT2A1外所有类固醇生成基因的转录。将存在和不存在福司可林时H295R细胞的转录谱与人肾上腺的转录谱进行了比较。与未处理细胞的表达模式相比，在福司可林处理的H295R细胞中观察到的基因表达模式与人肾上腺中的表达模式更相似。为了使用分子终点来检验H295R细胞作为评估肾上腺破坏作用的可能体外系统，使用了杀虫剂林丹（γ-六氯环己烷）。在体内，林丹已被证明可抑制睾丸、卵巢和肾上腺的类固醇生成。结果表明，林丹可降低皮质醇分泌，下调编码类固醇生成酶的一部分基因的表达，并抑制类固醇生成急性调节蛋白（StAR）基因启动子的转录激活。因此，H295R细胞系为在激素产生和基因表达水平分析人肾上腺类固醇生成途径提供了一个良好的体外系统。这种体外试验可用于快速检测肾上腺内分泌破坏作用，并作为进行机制研究的工具。
