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GATA-4 promotes the differentiation of P19 cells into cardiac myocytes.
Hu, De-Liang; Chen, Fu-Kun; Liu, Yao-Qiu; Sheng, Yan-Hui; Yang, Rong; Kong, Xiang-Qing; Cao, Ke-Jiang; Gu, Hai-Tao; Qian, Ling-Mei.
Afiliação
  • Hu DL; Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing, PR China.
Int J Mol Med ; 26(3): 365-72, 2010 Sep.
Article em En | MEDLINE | ID: mdl-20664952
The aim of this study was to investigate the effects of GATA-4 on the differentiation of P19 cells into cardiomyocytes and to examine the relationship between GATA-4 and cardiomyocytes. We constructed vectors to overexpress and silence GATA-4. These vectors, as well as empty ones were transfected into P19 cells. Subsequently, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were performed. The morphology of P19 cells during differentiation was observed using an inverted microscope. Total RNA was extracted from P19 cells. We used real-time PCR to evaluate the expression levels of 6 genes: GATA-4, GATA-6, transthyretin (TTR), alpha-fetoprotein (AFP), Nkx2.5, and alpha-myosin heavy chain (alpha-MHC). The gene expression pattern of these 6 genes is graphically shown for each group. The GATA-4 mRNA level in cells overexpressing GATA-4 was notably higher than that in the controls, whereas the levels in the controls were notably higher than those in the GATA-4-silenced P19 cells. The cell lines overexpressing GATA-4 expressed higher levels of Nkx2.5 and alpha-MHC than the controls. However, the controls expressed higher levels of AFP, GATA-6 and TTR than the cells overexpressing GATA-4. The RNAi group expressed lower levels of TTR, Nkx2.5, and alpha-MHC than the controls, but there were no differences in the RNAi group and the controls with regard to the expression levels of AFP and GATA-6. The gene expression pattern in the cells overexpressing GATA-4 was biased toward the Nkx2.5 and alpha-MHC. On the other hand, the gene expression pattern in GATA-4-silenced cells and the controls was biased toward the TTR and AFP. The overexpression of GATA-4 enhances the differentiation of P19 cells into cardiac myocytes, whereas its down-regulation suppresses this trend.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diferenciação Celular / Miócitos Cardíacos / Fator de Transcrição GATA4 Limite: Animals Idioma: En Revista: Int J Mol Med Assunto da revista: BIOLOGIA MOLECULAR / GENETICA MEDICA Ano de publicação: 2010 Tipo de documento: Article
Buscar no Google
Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diferenciação Celular / Miócitos Cardíacos / Fator de Transcrição GATA4 Limite: Animals Idioma: En Revista: Int J Mol Med Assunto da revista: BIOLOGIA MOLECULAR / GENETICA MEDICA Ano de publicação: 2010 Tipo de documento: Article