title: POD-1 binding to the E-box sequence inhibits SF-1 and StAR expression in human adrenocortical tumor cells.
abstract: Pod-1/Tcf21 is expressed at epithelial-mesenchymal interaction sites during development of many organs. Different approaches have demonstrated that Pod-1 transcriptionally inhibits Sf-1/NR5A1 during gonadal development. Disruption of Sf-1 can lead to disorders of adrenal development, while increased dosage of SF-1 has been related to increased adrenal cell proliferation and tumorigenesis. In this study, we analyzed whether POD-1 overexpression inhibits the endogenous Sf-1 expression in human and mouse adrenocortical tumor cells. Cells were transiently transfected with luciferase reporter gene under the control of Sf-1 promoter and with an expression vector encoding Pod-1. Pod-1 construct inhibited the transcription of the Sf1/Luc reporter gene in a dose-dependent manner in mouse Y-1 adrenocortical carcinoma (ACC) cells, and inhibited endogenous SF-1 expression in the human H295R and ACC-T36 adrenocortical carcinoma cells. These results were validated by chromatin immunoprecipitation assay with POD-1-transfected H295R cells using primers specific to E-box sequence in SF-1 promoter region, indicating that POD-1 binds to the SF-1 E-box promoter. Moreover, POD-1 over-expression resulted in a decrease in expression of the SF-1 target gene, StAR (Steroidogenic Acute Regulatory Protein). Lastly, while the induced expression of POD-1 did not affect the cell viability of H295R/POD-1 or ACC-T36/POD-1 cells, the most significantly enriched KEGG pathways for genes negatively correlated to POD-1/TCF21 in 33 human ACCs were those associated with cell cycle genes.
authors: Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900, SP, Brazil.
authors: Monica Malheiros
authors: MM
authors: França
authors: Bruno
authors: B
authors: Ferraz-de-Souza
authors: Mariza Gerdulo
authors: MG
authors: Santos
authors: Antonio Marcondes
authors: AM
authors: Lerario
authors: Maria Candida Barisson Villares
authors: MC
authors: Fragoso
authors: Ana Claudia
authors: AC
authors: Latronico
authors: Rork D
authors: RD
authors: Kuick
authors: Gary D
authors: GD
authors: Hammer
authors: Claudimara F P
authors: CF
authors: Lotfi
mesh_terms: D000306:Adrenal Cortex Neoplasms / Q000378:metabolism*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D024721:E-Box Elements; D006801:Humans; D051379:Mice; D010750:Phosphoproteins / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D054339:Steroidogenic Factor 1 / Q000096:biosynthesis*; D014158:Transcription, Genetic; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D010750:Phosphoproteins; D054339:Steroidogenic Factor 1; D000098682:Steroidogenic Acute Regulatory Protein; C515057:NR5A1 protein, human; C491825:TCF21 protein, human