Elementary studies on elevated steroidogenic factor-1 expression in aldosterone-producing adenoma

• Published in: Urologic oncology Vol. 30; no. 4; pp. 457 - 462
• Main Authors: Hu, Dongliang, M.D, Ouyang, Jinzhi, M.D, Wu, Zhun, M.D, Shi, Taoping, M.D, Wang, Baojun, M.D, Ma, Xin, M.D, Li, Hongzhao, M.D, Wang, Shaogang, M.D, Zhang, Xu, M.D
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.07.2012; Elsevier
• Subjects: Adrenal Cortex Neoplasms - genetics; Adrenal Cortex Neoplasms - metabolism; Adrenal Cortex Neoplasms - pathology; Adrenocortical Adenoma - genetics; Adrenocortical Adenoma - metabolism; Adrenocortical Adenoma - pathology; Aldosterone - biosynthesis; Aldosterone-producing adenoma; Apoptosis; Biological and medical sciences; Blotting, Western; Cell Line; Cell Proliferation; Gene Expression Regulation, Neoplastic; H295R cell; Humans; Immunohistochemistry; Ki-67 Antigen - analysis; Medical sciences; Microscopy, Fluorescence; Nephrology. Urinary tract diseases; Proliferation; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNAi; Steroidogenic Factor 1 - genetics; Steroidogenic Factor 1 - metabolism; Steroidogenic factor-1; Tumors; Urology; Steroidogenic factor-1; RNAi; Proliferation; Aldosterone-producing adenoma; H295R cell; Cell proliferation; Nephrology; RNA interference; Steroid hormone; Mineralocorticoid; Adenoma; Aldosterone; Urology; Gene silencing; Cancerology; Adrenal hormone; Benign neoplasm
• ISSN: 1078-1439; 1873-2496
• DOI: 10.1016/j.urolonc.2010.03.001

Abstract Objectives The expression of steroidogenic factor-1 (SF-1) was elevated in adrenal aldosterone-producing adenoma (APA). The influence of SF-1 on adrenal tumorigenesis by adrenocortical cell line H295R cells was investigated. Materials and methods Real-time PCR and Western blotting were used to detect SF-1 expression in 16 APA samples and 12 normal adrenal samples. Specific SF-1-shRNA plasmid was transfected into H295R cells to inhibit SF-1 expression. Western blotting and real-time PCR were used to verify the effects of RNAi on SF-1 inhibition. Subsequently, WST-1 and cell count were applied to evaluate cell proliferation at different SF-1 levels. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to measure cell apoptosis, and proliferation marker Ki-67 was studied by immunohistochemistry. Results Compared with normal adrenal samples, SF-1 mRNA and protein levels in APA samples were significantly higher. It was 10.48:1 at SF-1 mRNA and 0.87 ± 0.05 vs. 0.39 ± 0.07 at protein levels, respectively ( P < 0.01). A decreased SF-1 significantly inhibited cell proliferation in the experimental and control cells. These results were supported by weaker Ki-67 staining in SF-1-inhibited cells [(36.9% ± 4.17%) vs. (58.48% ± 7.16%) ( P < 0.01)]. Moreover, SF-1 inhibition induced a 2.7-fold increase in the percentage of apoptotic H295R cells ( P < 0.01). Conclusions Elevated SF-1 may play an important role in APA formation and primary aldosteronism. SF-1 acts as an oncogenic factor, and its inhibition provides new insight into the understanding and treatment of related adrenal diseases.