Session: SUN 17-28-Adrenal Tumors & Pheochromocytoma
Poster Board SUN-28
Monica Malheiros França1, Bruno Ferraz-de-Souza2, Mariza G. Santos3, Antonio M. Lerario3, Maria Candida B Villares Fragoso3, Ana Claudia Latronico3, Gary D. Hammer4, Claudimara F. Lotfi1
1Department of Anatomy, Biomedical Science Institute, 2Laboratory of Carbohydrates and Radioimmunoassays (LIM18); 3Laboratory of Molecular Genetics and Hormones (LIM42), School of Medicine; University of São Paulo, SP, Brazil, 4Department of Internal Medicine, Metabolism, Endocrinology and Diabetes, University of Michigan, USA.
Pod-1/Tcf21 has been showed to inhibit Sf-1 expression by antagonizing the activity of USF on the proximal E-box Sf-1 promoter site in Leydig cells. Also, an increase in Sf-1 expression was observed in Pod-1 deficient-Leydig cells. Recently, we have shown that POD-1 overexpression inhibits endogenous SF-1 expression through binding to the E-box sequence on SF-1 promoter in human adrenocortical tumor cells. Here, we detected cMYC and SF-1 protein in H295R and in an adult adrenocortical carcinoma cells (ACC-T36) transfected with pCMVMyc and pCMVMycPod-1. Furthermore, CYP11A1 gene expression was quantified in these H295R and ACC-T36 transfected cells. In addition, the expression of POD-1 and SF-1 was evaluated in pediatric adrenocortical tumor cells (ACT-T7 cells) transfected with siRNA targeting POD-1 by using qPCR. Our results show an increase of cMYC protein expression and a decrease of SF-1 protein in both H295R and ACC-T36 cells transfected with pCMVMycPod-1 in relation to control. Specifically we observed 0.77 ± 0.2 and 0.70 ± 0.1-fold decrease of CYP11A1 expression respectively in both cell lines when compared with the cells transfected with the empty plasmid. Preliminary results show 0.6-fold decrease of POD-1 expression and 9.2-fold increase of SF-1 expression in POD-1 knockdown of ACT-T7 cells transfected with siRNA-POD-1, in comparison with siRNAcontrol. In summary our results suggest a regulatory effect of POD-1 on SF-1 expression in both pediatric and adult adrenocortical tumor cells.
Nothing to Disclose: MMF, BF, MGS, AML, MCBVF, ACL, GDH, CFPL
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