Development of an Adrenocorticotropin-Responsive Human Adrenocortical Carcinoma Cell Line
Development of an Adrenocorticotropin-Responsive Human Adrenocortical Carcinoma Cell Line
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DOI：
10.1210/jc.2008-0903
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发表时间：
2008-11-01
影响因子：
5.8
通讯作者：
Rainey, William E.
中科院分区：
文献类型：
--
作者：
Parmar, Jeniel;Key, Rebecca E.;Rainey, William E.
背景：调节肾上腺类固醇生成的分子机制仍在研究中。目前唯一的人肾上腺皮质细胞系是NCI-H295及其亚株。其中一种菌株H295 R保留了对血管紧张素II（Ang II）的反应能力;然而，它缺乏ACTH反应性。一个促肾上腺皮质激素反应的人肾上腺皮质模型将增加显着的研究，旨在定义的分子控制corticosteroid biosynthesis.Objective：该研究的目的是开发一种人肾上腺细胞系，保留了血管紧张素II和促肾上腺皮质激素调节corticosteroid production.Design：人肾上腺皮质癌（HAC）细胞被分离出的肾上腺肿瘤从一个女孩提出男性化和高血压。建立并表征细胞的克隆群体。HAC细胞用ACTH，血管紧张素II，和毛喉素，然后通过检查类固醇生成酶mRNA的表达，使用定量实时PCR和类固醇production.Results：HAC克隆15（HAC 15）细胞反应治疗与ACTH，血管紧张素II，和毛喉素，增加皮质醇和醛固酮的生产。促肾上腺皮质激素、血管紧张素II和毛喉素还增加了mRNA的表达，这些mRNA编码皮质醇和醛固酮生物合成所需的所有酶，即类固醇生成急性调节蛋白、胆固醇侧链裂解酶、细胞色素P450 17 α-羟化酶-17，20-裂解酶、II型3 β-羟基类固醇脱氢酶、21-羟化酶、11 β-羟化酶和11 β-醛固酮合成酶。此外，细胞表达促肾上腺皮质激素受体（MC 2 R）和血管紧张素II受体的mRNA。MC 2 R蛋白也表达在HAC 15细胞中。结论：目前的研究描述了一个ACTH和Ang II反应的人肾上腺细胞系的发展和特点。HAC 15细胞系应提供一个重要的模型系统，以确定调节醛固酮和皮质醇生产的分子机制。（临床内分泌代谢杂志93：4542-4546，2008）
Context: The molecular mechanisms regulating adrenal steroidogenesis continue to be defined. The only current human adrenocortical cell line is the NCI-H295 and its substrains. One of the strains, H295R, has retained the ability to respond to angiotensin II (Ang II); however, it lacks ACTH responsiveness. An ACTH-responsive human adrenocortical model would add significantly to studies directed at defining the molecular control of corticosteroid biosynthesis.Objective: The objective of the study was to develop a human adrenal cell line that retained both Ang II-and ACTH-regulated corticosteroid production.Design: Human adrenocortical carcinoma (HAC) cells were isolated from an adrenal tumor removed from a girl presenting with virilization and hypertension. Clonal populations of cells were established and characterized. HAC cells were treated with ACTH, Ang II, and forskolin, followed by examination of steroidogenic enzyme mRNA expression using quantitative real-time PCR and steroid production.Results: HAC clone 15 (HAC15) cells responded to treatment with ACTH, Ang II, and forskolin, with increased cortisol and aldosterone production. ACTH, Ang II, and forskolin also increased expression of mRNA, encoding all enzymes needed for cortisol and aldosterone biosynthesis, namely steroidogenic acute regulatory protein, cholesterol side-chain cleavage, cytochrome P450 17 alpha-hydroxylase-17, 20-lyase, 3 beta-hydroxysteroid dehydrogenase type II, 21-hydroxylase, 11 beta-hydroxylase, and 11 beta-aldosterone synthase. In addition, the cells expressed mRNA for ACTH receptor (MC2R) and Ang II receptor. MC2R protein was also expressed in HAC15 cells.Conclusion: The current study describes the development and characterization of an ACTH- and Ang II-responsive human adrenal cell line. The HAC15 cell line should provide an important model system for defining the molecular mechanisms regulating aldosterone and cortisol production. (J Clin Endocrinol Metab 93: 4542-4546, 2008)