Role of COUP-TFI during retinoic acid-induced differentiation of P19 cells to endodermal cells

• Published in: Journal of cellular physiology 2013-04, Vol.228 (4), p.791-800
• Main Authors: Pickens, Brandy S., Teets, Bryan W., Soprano, Kenneth J., Soprano, Dianne Robert
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis - drug effects; Apoptosis - genetics; Cell Cycle - drug effects; Cell Cycle - genetics; Cell differentiation; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cells, Cultured; COUP Transcription Factor I - genetics; COUP Transcription Factor I - metabolism; COUP Transcription Factor II - genetics; COUP Transcription Factor II - metabolism; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; DAX-1 Orphan Nuclear Receptor - genetics; DAX-1 Orphan Nuclear Receptor - metabolism; Endoderm - cytology; Endoderm - drug effects; Endoderm - metabolism; Gene Expression - drug effects; Gene Expression - genetics; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Leukemia; Mice; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - metabolism; Pre-B-Cell Leukemia Transcription Factor 1; Retinoic Acid 4-Hydroxylase; RNA, Messenger - genetics; Steroidogenic Factor 1 - genetics; Steroidogenic Factor 1 - metabolism; Thyroid; Transcription Factors - genetics; Transcription Factors - metabolism; Tretinoin - pharmacology
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.24228

Retinoic acid (RA) is a positive regulator of P19 cell differentiation. Silencing of pre‐B cell leukemia transcription factors (PBXs) expression in P19 cells (AS cells) results in a failure of these cells to differentiate to endodermal cells upon RA treatment. Chicken Ovalbumin Upstream Promoter Transcription Factor I (COUP‐TFI) is an orphan member of the steroid‐thyroid hormone superfamily. RA treatment of wild type P19 cells results in a dramatic increase in the expression of COUP‐TFI; however, COUP‐TFI mRNA levels fail to be elevated upon RA treatment of AS cells indicating that PBX expression is required for elevation in COUP‐TFI expression. To study the role of COUP‐TFI during RA‐dependent differentiation of P19 cells, AS cells that inducibly express various levels of COUP‐TFI were prepared. Exogenous expression of COUP‐TFI in AS cells, in a dose‐dependent fashion, leads to growth inhibition, modest cell cycle disruption, and early apoptosis. Furthermore, AS cells can overcome the blockage in RA‐dependent differentiation to endodermal cells when either pharmacological levels of COUP‐TFI are expressed or a combination of both the expression of physiological levels of COUP‐TFI and RA treatment. Additionally, the mRNA level of several pluripotency associated genes including OCT‐4, DAX‐1, and SF‐1 in the COUP‐TFI expressing AS cells are reduced. Moreover, analysis of the expression of primary RA response genes indicates that COUP‐TFI is involved in the regulatory modulation of the expression of at least two genes, CYP26A1 and HoxA1. These studies demonstrate that COUP‐TFI functions as a physiologically relevant regulator during RA‐mediated endodermal differentiation of P19 cells. J. Cell. Physiol. 228: 791–800, 2013. © 2012 Wiley Periodicals, Inc.