Adenovirus-mediated expression of hypoxia-inducible factor 1 alpha double mutant converts neonatal cardiac fibroblasts into (cardio)myocyte phenotype

作者：Wang, Yesong^*; Sun, Aijiao; Xue, Jiaojie; Jiang, Yi

来源：Cell Biochemistry and Function, 2012, 30(1): 24-32.

DOI：10.1002/cbf.1813

• fibroblasts
• hypoxia-inducible factor-1a (HIF-1a)
• mutant
• adenovirus
• vector
• phenotype

Adenovirus-mediated expression of hypoxia-inducible factor 1a double mutant (pAd-HIF-1a-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 1a (HIF-1a), 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-1a-Ala564-Ala803 also can convert neonatal rat cardiac fibroblasts (NCFs) into (cardio)myocyte phenotype via regulating the downstream target genes of HIF-1a at normoxia. The immunostaining analysis showed that NCFs treated with pAd-HIF-1a-Ala564-Ala803 exhibited higher protein expression levels of smooth muscle a-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-1a-Ala564-Ala803 augmented messenger RNA (mRNA) expression of transforming growth factor-beta 1 (TGF-beta 1), Smad4, NKx2.5, GATA4, myocardin, SMA and cTnT. The effects of HIF-1a-Ala564-Ala803 on NCFs were attenuated by pre-transfection of TGF-beta 1 or myocardin small interference RNAs. Adult CFs transfected with pAd-HIF-1a-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-1a-Ala564-Ala803 can convert NCFs into (cardio)myocyte phenotype via regulating its downstream target genes.

• 出版日期2012-1
• 单位中山大学