Characterization of NCI-H295R Cells as an In Vitro Model of Hyperaldosteronism
Characterization of NCI-H295R Cells as an In Vitro Model of Hyperaldosteronism
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DOI：
10.1055/s-0032-1323810
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发表时间：
2013-02-01
影响因子：
2.2
通讯作者：
Beuschlein, F.
中科院分区：
文献类型：
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作者：
Lichtenauer, U. D.;Shapiro, I.;Beuschlein, F.
由于缺乏反映产生醛固酮的腺瘤功能特征的肿瘤细胞系，对醛固酮分泌功能自主性的关键分子机制的深入分析受到阻碍。在此，我们描述了肾上腺皮质癌细胞系NCI-H295R的特性，以及它在不同培养条件下作为醛固酮增多症模型的适用性。类固醇图谱显示，NCI-H295R细胞主要分泌皮质醇，而醛固酮和其他类固醇的释放浓度要低得多。然而，不同的刺激如ACTH和血管紧张素II，尤其是钾以剂量依赖的方式增加了醛固酮的产量。NCI-H295R细胞在特定的培养条件下很容易形成球体，这是一种广泛用于扩增祖细胞的方法。出乎意料的是，球形细胞具有更高的醛固酮浓度和更高的类固醇生成酶STAR、3βHSD、CYP17、SF-1和MC2受体的表达水平。进一步的研究表明，这种现象主要归因于上皮生长因子(EGF)，特别是成纤维细胞生长因子(FGF)，这两种成分都是球形培养基中的必需成分。联合作用下的醛固酮释放量并不高于单独应用成纤维细胞生长因子的作用。因此，球体生长本身并不能确保该细胞系中分化较少的细胞类型的丰富。
In depth analysis of key molecular mechanisms involved in functional autonomy of aldosterone secretion is hampered by the lack of tumor cell lines that reflect functional characteristics of aldosterone producing adenomas. Herein, we describe the characteristics of the adrenocortical carcinoma cell line NCI-H295R and its suitability as a model of hyperaldosteronism in relation to different culture conditions. Steroid profiling revealed that NCI-H295R cells predominantly secrete cortisol, while aldosterone and other steroids are released at much lower concentrations. However, aldosterone output specifically increased in response to different stimuli such as ACTH and angiotensin II, and in particular to potassium in a dose dependent manner. NCI-H295R cells readily formed spheroids under specific culture conditions, a method widely used for the enrichment of progenitor cells. Unexpectedly, spheroid cells excelled with higher aldosterone concentration and higher expression levels of the steroidogenic enzymes StAR, 3 beta HSD, CYP17, SF-1, and the MC2-receptor. Further investigations revealed that this phenomenon is mainly attributed to epithelial growth factor (EGF) and particularly fibroblast growth factor (FGF), which are both essential ingredients in the spheroid culture medium. Aldosterone release under the combinatory influence of EGF and FGF was not higher than the effect of FGF alone. Spheroid growth per se, therefore, does not ensure an enrichment of less differentiated cell types in this cell line.