Studies on wnt5a inducing differentiation of cardiac progenitor cells to myocardial cells / 解放军医学杂志

ABSTRACT

Objective To investigate the effect of Wnt5a on inducing differentiation of Cp15-5a cell to myocardial cell.Methods Recombinant adenovirus wnt5a (Ad-wnt5a) and Ad-GFP was amplified with human embryo kidney 293 cells (HEK293 cells),and then transfected into CP15-5a cells and 3 experiment groups were set upwnt5a group,GFP group and blank control group.Flow cytometry was used to detect the transfection efficiency of Ad-wnt5a and Ad-GFP.One week after transfection,the expressions of genes GATA binding protein 4 (GATA4) and myocardial enhancement factor 2C (MEF2C) were analyzed by realtime quantitative PCR (qRT-PCR).Two weeks after transfection,the expressions of cardiac-specific connexin 43 (Cx43) and cardiac troponin T (cTnT) in Ad-wnt5a-induced CP15-5a cells were detected by Western blotting and immunofluorescence techniques.The sodium current expression (INa) was detected by whole cell patch clamp techniques.Results The transfection efficiency of Ad-wnt5a and Ad-GFP was 42.8％ and 44.3％,respectively.One week after transduction,the expressions of GATA4 and MEF2C were significantly higher in wnt5a group (1.717 ± 0.220 and 1.847 ± 0.190) than in GFP group (1.003 ± 0.087 and 0.456 ± 0.042,P＜0.05) and blank control group (0.961 ± 0.063 and 0.500 ± 0.095,P＜0.05),while no significant difference existed between GFP group and blank control group.Two weeks after transduction,the expressions of CX43 and cTnT were significantly higher in wnt5a group (1.597 ± 0.267 and 0.727 ± 0.100) than in GFP group (0.723 ± 0.047 and 0.217 ± 0.021,P＜0.05) and blank control group (0.783 ± 0.1333 and 0.253 ± 0.102,P＜0.01),while no significant difference existed between GFP group and blank control group.INa was detected in the wnt5a group compared with GFP group and blank control group.Conclusion wnt5a may induce differentiation of cp 15-5a cell into myocardial cell.