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Development of an adrenocorticotropin-responsive human adrenocortical carcinoma cell line
Jeniel Parmar1, Rebecca E Key, William E Rainey
1Department of Physiology, Medical College of Georgia, Augusta, Georgia 30912, USA.
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 17alpha-hydroxylase-17, 20-lyase, 3beta-hydroxysteroid dehydrogenase type II, 21-hydroxylase, 11beta-hydroxylase, and 11beta-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.
Insights
Researchers developed a new human adrenal cell line (HAC15) that responds to ACTH and Ang II. This model aids in studying molecular mechanisms of cortisol and aldosterone production.
Area of Science:
- Endocrinology
- Molecular Biology
- Cell Biology
Background:
- Adrenal steroidogenesis regulation is complex and not fully understood.
- Existing human adrenocortical cell lines, like H295R, have limitations, lacking ACTH responsiveness.
- A dual-responsive model is crucial for studying corticosteroid biosynthesis control.
Purpose of the Study:
- To develop and characterize a novel human adrenal cell line responsive to both ACTH and Ang II.
- To establish a reliable model for investigating the molecular regulation of steroidogenesis.
Main Methods:
- Human adrenocortical carcinoma (HAC) cells were isolated from a tumor.
- Clonal cell populations were established and characterized.
- Cells were treated with ACTH, Ang II, and forskolin, followed by analysis of steroidogenic enzyme mRNA and steroid production.
Main Results:
- HAC clone 15 (HAC15) cells demonstrated increased cortisol and aldosterone production in response to ACTH, Ang II, and forskolin.
- Upregulation of mRNA for key steroidogenic enzymes (StAR, CYP11A1, CYP17A1, HSD3B2, CYP21A2, CYP11B1, CYP11B2) was observed.
- HAC15 cells expressed mRNA and MC2R protein for ACTH receptors and Ang II receptors.
Conclusions:
- A novel human adrenal cell line, HAC15, responsive to both ACTH and Ang II, has been successfully developed and characterized.
- The HAC15 cell line serves as a valuable model for elucidating molecular mechanisms governing cortisol and aldosterone synthesis.
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