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Adenovirus‐mediated expression of hypoxia‐inducible factor 1 α double mutant converts neonatal cardiac fibroblasts into (cardio)myocyte phenotype
Author(s) -
Wang Yesong,
Sun Aijiao,
Xue Jiaojie,
Jiang Yi
Publication year - 2012
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.1813
Subject(s) - myocardin , gata4 , transfection , biology , myocyte , microbiology and biotechnology , mesenchymal stem cell , transcription factor , immunostaining , cellular differentiation , small interfering rna , serum response factor , cell culture , immunology , gene , immunohistochemistry , genetics , biochemistry
Adenovirus‐mediated expression of hypoxia‐inducible factor 1 α double mutant (pAd‐HIF‐1 α ‐Ala564‐Ala803) can be effectively transfected into bone marrow stem cells (MSCs) in the MSCs and cardiomyocytes co‐culture system at normoxia to regulate the expression of downstream target genes of hypoxia‐inducible factor 1 α (HIF‐1 α ), which in turn can promote MSC differentiation into cardiomyocytes. Fibroblasts share common characteristics with MSCs such as the morphology, phenotype and differentiation potential. Therefore, we further studied whether the pAd‐HIF‐1 α ‐Ala564‐Ala803 also can convert neonatal rat cardiac fibroblasts (NCFs) into (cardio)myocyte phenotype via regulating the downstream target genes of HIF‐1α at normoxia. The immunostaining analysis showed that NCFs treated with pAd‐HIF‐1 α ‐Ala564‐Ala803 exhibited higher protein expression levels of smooth muscle α ‐actin (SMA, myocyte marker) and cardiac troponin T (cTnT, cardiomyocyte marker), compared with phosphate‐buffered saline and pAd‐LacZ treatments. The reverse transcription‐polymerase chain reaction results showed that NCFs transfected with pAd‐HIF‐1 α ‐Ala564‐Ala803 augmented messenger RNA (mRNA) expression of transforming growth factor‐ β 1 (TGF‐ β 1), Smad4, NKx2.5, GATA4, myocardin, SMA and cTnT. The effects of HIF‐1 α ‐Ala564‐Ala803 on NCFs were attenuated by pre‐transfection of TGF‐ β 1 or myocardin small interference RNAs. Adult CFs transfected with pAd‐HIF‐1 α ‐Ala564‐Ala803 showed a lower protein expression of SMA but not cTnT without any change in the mRNA expression level of NKx2.5, myocardin. Therefore, NCFs but not adult CFs possess a similar differentiation potential to MSCs as evidenced by the fact that pAd‐HIF‐1 α ‐Ala564‐Ala803 can convert NCFs into (cardio)myocyte phenotype via regulating its downstream target genes. Copyright © 2011 John Wiley & Sons, Ltd.