Critical role for transcription factor AP-2alpha in human trophoblast differentiation.
To examine whether AP-2alpha is a critical component of the genetic program that directs human trophoblast differentiation, we used DNA microarray analyses to characterize the effects of a dominant-negative form of the AP-2 protein upon in vitro differentiating cytotrophoblast cells. Human cytotrophoblast cells (>95% pure) were cultured for 3 days in the presence of control medium or medium containing an adenovirus that expresses a dominant-negative mutant of AP-2 (Ad2.AP-2D/N) or an adenovirus lacking the AP-2 mutant gene (Ad.WT). DNA microarray analyses using Affymetrix human U95Av2 GeneChips were performed on RNA extracted from the three groups of cells immediately prior to and after 3 days of cell culture. Cells infected with Ad2.AP-2D/N or Ad2.WT underwent morphological differentiation similar to that of uninfected cells, with greater than 90% of the cells in each group fusing to form multinucleated syncytiotrophoblast cells. However, Ad2.AP-2D/N markedly inhibited the induction or repression of many genes that were regulated in the noninfected and Ad2.WT-infected cells during differentiation. Eighteen of the 25 most induced genes and 17 of the 20 most repressed genes during differentiation were AP-2 dependent, with the majority of these related to extracellular organization, cellular communication, and signal transduction. Taken together, these findings strongly suggest that AP-2 plays a critical role for both the induction and repression of genes that comprise postsyncytialization gene expression programs of trophoblast differentiation and maturation. AP-2, however, is not required for the fusion of cytotrophoblast cells to form a syncytium or the expression of syncytin.
Cheng
You-Hong
YH
Department of Endocrinology, Children's Hospital Research Foundation and University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA. cheny0@cchmc.org
Aronow
Bruce J
BJ
Hossain
Shaikh
S
Trapnell
Bruce
B
Kong
Sue
S
Handwerger
Stuart
S
D002450:Cell Communication / Q000235:genetics; D002454:Cell Differentiation; D002455:Cell Division / Q000235:genetics; D002459:Cell Fusion; D002478:Cells, Cultured / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D015686:Gene Products, env / Q000096:biosynthesis / Q000235:genetics; D015726:Giant Cells; D006728:Hormones / Q000378:metabolism; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000096:biosynthesis / Q000235:genetics; D011263:Pregnancy Trimester, Third; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins; D015398:Signal Transduction / Q000235:genetics; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism
D004268:DNA-Binding Proteins; D015686:Gene Products, env; D006728:Hormones; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C493673:TFAP2A protein, human; D050656:Transcription Factor AP-2; D014157:Transcription Factors; C405335:syncytin