miR-200c and GATA binding protein 4 regulate human embryonic stem cell renewal and differentiation

详细信息    查看全文

• 作者：Hsiao-Ning Huang^a ; ^b ; Shao-Yin Chen^a ; Shiaw-Min Hwang^c ; Ching-Chia Yu^a ; Ming-Wei Su^d ; ^e ; Wei Mai^d ; ^e ; Hsei-Wei Wang^f ; ^g ; ^h ; ⁱ ; ^j ; Wei-Chung Cheng^f ; ^k ; Scott C. Schuyler^l ; Nianhan Ma^m ; Frank Leigh Luⁿ ; Jean Lu^a ; ^b ; ^d ; ^h ; ^o ; ^{jeanlu@gate.sinica.edu.tw" class="auth_mail}
• 关键词：hESCs ; human embryonic stem cells ; EB ; embryoid body ; TGF-尾 ; transforming growth factor-尾 ; FGF ; fibroblast growth factor ; miRNAs ; microRNAs ; qRT-PCR ; quantitative reverse transcription polymerase chain reaction.
• 刊名：Stem Cell Research
• 出版年：March, 2014
• 年：2014
• 卷：12
• 期：2
• 页码：338-353
• 全文大小：2586 K

文摘

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鈥?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-尾 (TGF-尾飥?/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.