title: miR-200c and GATA binding protein 4 regulate human embryonic stem cell renewal and differentiation.
abstract: Human embryonic stem cells (hESCs) are functionally unique for their self-renewal ability and pluripotency, but the molecular mechanisms giving rise to these properties are not fully understood. hESCs can differentiate into embryoid bodies (EBs) containing ectoderm, mesoderm, and endoderm. In the miR-200 family, miR-200c was especially enriched in undifferentiated hESCs and significantly downregulated in EBs. The knockdown of the miR-200c in hESCs downregulated Nanog expression, upregulated GATA binding protein 4 (GATA4) expression, and induced hESC apoptosis. The knockdown of GATA4 rescued hESC apoptosis induced by downregulation of miR-200c. miR-200c directly targeted the 3'-untranslated region of GATA4. Interestingly, the downregulation of GATA4 significantly inhibited EB formation in hESCs. Overexpression of miR-200c inhibited EB formation and repressed the expression of ectoderm, endoderm, and mesoderm markers, which could partially be rescued by ectopic expression of GATA4. Fibroblast growth factor (FGF) and activin A/nodal can sustain hESC renewal in the absence of feeder layer. Inhibition of transforming growth factor-β (TGF-β[Symbol: see text])/activin A/nodal signaling by SB431542 treatment downregulated the expression of miR-200c. Overexpression of miR-200c partially rescued the expression of Nanog/phospho-Smad2 that was downregulated by SB431542 treatment. Our observations have uncovered novel functions of miR-200c and GATA4 in regulating hESC renewal and differentiation.
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei, Taiwan; Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Hsiao-Ning
authors.initials: HN
authors.lastname: Huang
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Shao-Yin
authors.initials: SY
authors.lastname: Chen
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan.
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Ching-Chia
authors.initials: CC
authors.lastname: Yu
authors.affiliation: National RNAi Platform/National Core Facility Program for Biotechnology, Taipei, Taiwan; Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
authors.forename: Ming-Wei
authors.initials: MW
authors.lastname: Su
authors.affiliation: National RNAi Platform/National Core Facility Program for Biotechnology, Taipei, Taiwan; Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
authors.forename: Wei
authors.initials: W
authors.lastname: Mai
authors.affiliation: VGH-YM Genomic Research Center, National Yang-Ming University, Taipei, Taiwan; Institute of Biomedical Informatics, National Yang-Ming University, Taipei, Taiwan; Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan; Cancer Research Center, National Yang-Ming University, Taipei, Taiwan; Department of Education and Research, Taipei City Hospital, Taipei, Taiwan.
authors.forename: Hsei-Wei
authors.initials: HW
authors.lastname: Wang
authors.affiliation: VGH-YM Genomic Research Center, National Yang-Ming University, Taipei, Taiwan; Division of Pediatric Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Wei-Chung
authors.initials: WC
authors.lastname: Cheng
authors.affiliation: Department of Biomedical Science, College of Medicine, Chang Gung University, Taoyuan County, Taiwan.
authors.forename: Scott C
authors.initials: SC
authors.lastname: Schuyler
authors.affiliation: Institute of Systems Biology and Bioinformatics, National Central University, Taoyuan, Taiwan.
authors.forename: Nianhan
authors.initials: N
authors.lastname: Ma
authors.affiliation: Department of Pediatrics, National Taiwan University Hospital, National Taiwan University Medical College, Taipei, Taiwan.
authors.forename: Frank Leigh
authors.initials: FL
authors.lastname: Lu
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei, Taiwan; Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan; National RNAi Platform/National Core Facility Program for Biotechnology, Taipei, Taiwan; Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan; Genomics and System Biology Program, College of Life Science, National Taiwan University, Taipei, Taiwan. Electronic address: jeanlu@gate.sinica.edu.tw.
authors.forename: Jean
authors.initials: J
authors.lastname: Lu
mesh_terms: D028341:Activins / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D019070:Cell Lineage; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D050991:GATA4 Transcription Factor; C484170:GATA4 protein, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D016212:Transforming Growth Factor beta; C422405:activin A; D028341:Activins