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- W2364156312 abstract "Objective To investigate the effects of down-regulation of desmocollin-2( DSC2) gene expression on cardiac-like myocyte differentiated from P19 cells. Methods The DSC2 gene specific siRNAs and control siRNA( siNC)were synthesized and cloned into pGPU6 /GFP /Neo vector. The recombinant plasmids( ShDSC2 and ShNC) were transfected into P19 cells,and the stable cell lines were selected with G418. The efficiency of DSC2 knock down at the levels of mRNA and protein expression were determined by RT-PCR and Western blot,respectively. The selected P19 cells lines were then induced and differentiated into cardiomyocyte-like cells by DMSO. The apoptosis process and ultrastructure changes of cardiomyocyte-like cell derived from P19 cells were evaluated by flow cytometry and transmission electron microscope( TEM),respectively. RT-PCR was used to measure the expression of adipogenesis and fibrogenesis related genes at mRNA level. Results Five recombinant ShDSC2 plasmids were constructed successfully and transfected into P19 cells,after G418 selection,positive clones were established by RNA interference. Compared with the ShNC control group,the expression levels of mRNA and protein in ShDSC2 group were decreased by 69. 47%( P0. 01) and 65. 62%( P0. 01),respectively. In ShDSC2 group,TEM showed that fat droplet,cellular vacuoles,swollen mitochondria and disappeared cristae,flow cytometry demonstrated that the apoptosis was also increased. A marked increase in expression levels of three major regulators of adipogenesis,namely PPAR-γ C /EBP-α and Adiponectin,and their target gene adiponectin,were also observed in ShDSC2 group. The expression levels of procollagen genes Col1a1,Col1a2 and Col3a1 were increased remarkably. Conclusions We established DSC2 gene silenced P19 cell line and further differentiated it into cardiomyocyte-like cells. The later exhibited increased apoptosis,adipogenesis and fibrogenesis,thus recapitulating the phenotype of human arrhythmogenic right ventricular cardiomyopathy.This cellular model could provide the opportunity to explore the pathogenesis and novel tool for therapeutic studies of ARVC in vitro." @default.
- W2364156312 created "2016-06-24" @default.
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- W2364156312 date "2014-01-01" @default.
- W2364156312 modified "2023-09-23" @default.
- W2364156312 title "The effects of silencing desmocollin-2 gene on the cardiomyocyte-like cells differentiated from P19 cell lines" @default.
- W2364156312 hasPublicationYear "2014" @default.
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