Involvement of Transcription Factor NR2F2 in Human Trophoblast Differentiation

Background During the in vitro differentiation of human villous cytotrophoblast (CTB) cells to a syncytiotrophoblast (STB) phenotype, mRNA levels for the nuclear hormone receptor NR2F2 (ARP-1, COUP-TFII) increase rapidly, reaching a peak at day 1 of differentiation that is 8.8-fold greater than that in undifferentiated CTB cells. To examine whether NR2F2 is involved in the regulation of villous CTB cell differentiation, studies were performed to determine whether NR2F2 regulates the expression of TFAP2A (AP-2α), a transcription factor that is critical for the terminal differentiation of these cells to a STB phenotype. Methodology/Primary Findings Overexpression of NR2F2 in primary cultures of human CTB cells and JEG-3 human choriocarcinoma cells induced dose-dependent increases in TFAP2A promoter activity. Conversely, siRNA mediated silencing of the NR2F2 gene in villous CTB undergoing spontaneous differentiation blocked the induction of the mRNAs for TFAP2A and several STB cell specific marker genes, including human placental lactogen (hPL), pregnancy specific glycoprotein 1 (PSG1) and corticotropin releasing hormone (CRH) by 51–59%. The induction of TFAP2A promoter activity by NR2F2 was potentiated by the nuclear hormone receptors retinoic acid receptor alpha (RARA) and retinoid X receptor alpha (RXRA). Conclusions/Significance Taken together, these results strongly suggest that NR2F2 is involved in villous CTB cell differentiation and that NR2F2 acts, at least in part, by directly activating TFAP2A gene expression and by potentiating the transactivation of TFAP2A by RARA and RXRA.

[1]  M. Tsai,et al.  Deletion of the orphan nuclear receptor COUP-TFII in uterus leads to placental deficiency , 2007, Proceedings of the National Academy of Sciences.

[2]  M. Tsai,et al.  Molecular mechanisms involved in progesterone receptor regulation of uterine function , 2006, The Journal of Steroid Biochemistry and Molecular Biology.

[3]  B. Huppertz,et al.  Trophoblast fusion: fusogenic proteins, syncytins and ADAMs, and other prerequisites for syncytial fusion. , 2006, Micron.

[4]  F. Kikkawa,et al.  Ikaros is expressed in human extravillous trophoblasts and involved in their migration and invasion. , 2005, Molecular human reproduction.

[5]  S. Handwerger,et al.  A Placenta-Specific Enhancer of the Human Syncytin Gene1 , 2005, Biology of reproduction.

[6]  Qiang Wang,et al.  The evidences of human orphan receptor COUP-TFII inhibiting telomerase activity through decreasing hTERT transcription. , 2004, Cancer letters.

[7]  B. Aronow,et al.  Critical role for transcription factor AP-2alpha in human trophoblast differentiation. , 2004, Physiological genomics.

[8]  C. Peng.  The TGF-β Superfamily and Its Roles in the Human Ovary and Placenta , 2003 .

[9]  S. Mizutani,et al.  Placental leucine aminopeptidase/oxytocinase gene regulation by activator protein‐2 in BeWo cell model of human trophoblast differentiation , 2003, FEBS letters.

[10]  Roger A. Davis,et al.  ARP-1/COUP-TF II Determines Hepatoma Phenotype by Acting as Both a Transcriptional Repressor of Microsomal Triglyceride Transfer Protein and an Inducer of CYP7A1* , 2003, Journal of Biological Chemistry.

[11]  C. Peng.  The TGF-beta superfamily and its roles in the human ovary and placenta. , 2003, Journal of obstetrics and gynaecology Canada : JOGC = Journal d'obstetrique et gynecologie du Canada : JOGC.

[12]  S. Handwerger,et al.  AP-2α modulates human corticotropin-releasing hormone gene expression in the placenta by direct protein–protein interaction , 2002, Molecular and Cellular Endocrinology.

[13]  S. Handwerger,et al.  AP-2alpha modulates human corticotropin-releasing hormone gene expression in the placenta by direct protein-protein interaction. , 2002, Molecular and cellular endocrinology.

[14]  B. Aronow,et al.  Microarray analysis of trophoblast differentiation: gene expression reprogramming in key gene function categories. , 2001, Physiological genomics.

[15]  C. Maccalman,et al.  α‐, β‐, γ‐catenin, and p120CTN expression during the terminal differentiation and fusion of human mononucleate cytotrophoblasts in vitro and in vivo , 2001 .

[16]  C. Maccalman,et al.  alpha-, beta-, gamma-catenin, and p120(CTN) expression during the terminal differentiation and fusion of human mononucleate cytotrophoblasts in vitro and in vivo. , 2001, Molecular reproduction and development.

[17]  Y. Sadovsky,et al.  Peroxisome proliferator-activated receptor-γ modulates differentiation of human trophoblast in a ligand-specific manner , 2000 .

[18]  Takashi Suzuki,et al.  Immunohistochemical distribution of chicken ovalbumin upstream promoter transcription factor II in human tissues , 2000, Molecular and Cellular Endocrinology.

[19]  C. Maccalman,et al.  Regulation of beta-catenin mRNA and protein levels in human villous cytotrophoblasts undergoing aggregation and fusion in vitro: correlation with E-cadherin expression. , 2000, Journal of reproduction and fertility.

[20]  C. Kessler,et al.  Activator protein-2 regulates human placental lactogen gene expression , 2000, Molecular and Cellular Endocrinology.

[21]  J. Mccoy,et al.  Syncytin is a captive retroviral envelope protein involved in human placental morphogenesis , 2000, Nature.

[22]  J. Chiang,et al.  HNF4 and COUP-TFII interact to modulate transcription of the cholesterol 7alpha-hydroxylase gene (CYP7A1). , 2000, Journal of lipid research.

[23]  S. Smith,et al.  The Journal of Clinical Endocrinology & Metabolism Printed in U.S.A. Copyright © 2000 by The Endocrine Society Peroxisome Proliferator-Activated Receptor- � Modulates Differentiation of Human Trophoblast in a Ligand-Specific Manner* , 2022 .

[24]  F. Soncin,et al.  Retinoid receptors expression in human term placenta: involvement of RXR alpha in retinoid induced-hCG secretion. , 1998, The Journal of clinical endocrinology and metabolism.

[25]  F. Soncin,et al.  Retinoid Receptors Expression in Human Term Placenta: Involvement of RXRα in Retinoid Induced-hCG Secretion , 1998 .

[26]  G. Salbert,et al.  The nuclear orphan receptors COUP-TF and ARP-1 positively regulate the trout estrogen receptor gene through enhancing autoregulation , 1997, Molecular and cellular biology.

[27]  K. Nakao,et al.  Nonadipose tissue production of leptin: Leptin as a novel placenta-derived hormone in humans , 1997, Nature Medicine.

[28]  B. Paulweber,et al.  Functional domains of the human orphan receptor ARP-1/COUP-TFII involved in active repression and transrepression , 1997, Molecular and cellular biology.

[29]  S. Handwerger,et al.  Regulation of the Human Chorionic Gonadotropin α- and β-Subunit Promoters by AP-2* , 1997, The Journal of Biological Chemistry.

[30]  S. Handwerger,et al.  The ARP-1 orphan receptor represses steroid-mediated stimulation of human placental lactogen gene expression. , 1996, Journal of molecular endocrinology.

[31]  S. Cereghini,et al.  Positive regulation of the vHNF1 promoter by the orphan receptors COUP-TF1/Ear3 and COUP-TFII/Arp1 , 1996, Molecular and cellular biology.

[32]  R. Vihko,et al.  Coordination of transcription of the human 17 beta-hydroxysteroid dehydrogenase type 1 gene (EDH17B2) by a cell-specific enhancer and a silencer: identification of a retinoic acid response element. , 1995, Molecular endocrinology.

[33]  N. Harada,et al.  Regulation of Placenta-specific Expression of the Aromatase Cytochrome P-450 Gene , 1995, The Journal of Biological Chemistry.

[34]  Y. Kaneko,et al.  Molecular interactions during pregnancy: Expression of leukaemia inhibitory factor (LIF) receptor in human placenta: a possible role for LIF in the growth and differentiation of trophoblasts , 1995 .

[35]  Y. Kaneko,et al.  Expression of leukaemia inhibitory factor (LIF) receptor in human placenta: a possible role for LIF in the growth and differentiation of trophoblasts. , 1995, Human Reproduction.

[36]  S. Karathanasis,et al.  Transcriptional activation by the orphan nuclear receptor ARP-1. , 1995, Nucleic acids research.

[37]  H. Matsuo,et al.  Insulin-like growth factor-I as a local regulator of proliferation and differentiated function of the human trophoblast in early pregnancy. , 1995, Early pregnancy : biology and medicine : the official journal of the Society for the Investigation of Early Pregnancy.

[38]  J. Whelan,et al.  Commonly used cat reporter vectors contain a cAMP-inducible, cryptic enhancer that co-operates with NF-kappa B-sites. , 1994, Gene.

[39]  D. Morrish,et al.  Demonstration of functional cytokine-placental interactions: CSF-1 and GM-CSF stimulate human cytotrophoblast differentiation and peptide hormone secretion. , 1994, Experimental cell research.

[40]  J. Baxter,et al.  Eukaryotic regulatory elements lurking in plasmid DNA: the activator protein-1 site in pUC. , 1994, Molecular endocrinology.

[41]  J. Chou,et al.  Characterization of upstream activation elements essential for the expression of germ cell alkaline phosphatase in human choriocarcinoma cells. , 1993, The Journal of biological chemistry.

[42]  J. Lin,et al.  Novel role of human chorionic gonadotropin in differentiation of human cytotrophoblasts. , 1993, Endocrinology.

[43]  H. Matsuo,et al.  [Epidermal growth factor regulates trophoblast proliferation and differentiation in an autocrine/paracrine manner]. , 1993, Nihon Sanka Fujinka Gakkai zasshi.

[44]  J. Strauss,et al.  The cAMP signalling system and human trophoblast function. , 1992, Placenta.

[45]  K. Umesono,et al.  Convergence of 9-cis retinoic acid and peroxisome proliferator signalling pathways through heterodimer formation of their receptors , 1992, Nature.

[46]  S. Karathanasis,et al.  Repression by ARP-1 sensitizes apolipoprotein AI gene responsiveness to RXR alpha and retinoic acid , 1992, Molecular and cellular biology.

[47]  M. Pfahl,et al.  Retinoid X receptor is an auxiliary protein for thyroid hormone and retinoic acid receptors , 1992, Nature.

[48]  T. Wagner,et al.  Fertility of transgenic female mice expressing bovine growth hormone or human growth hormone variant genes. , 1991, Biology of reproduction.

[49]  G. Braunstein,et al.  Retinoic acid stimulates placental hormone secretion by choriocarcinoma cell lines in vitro. , 1991, Endocrinology.

[50]  R. Tjian,et al.  Regulation of transcription factor AP-2 by the morphogen retinoic acid and by second messengers. , 1989, Genes & development.