Up-regulation of PKC and during beating cardiomyocyte differentiation of P19CL6 cells with suppressed apoptotic cell populations

BackgroundCardiomyocyte differentiation has been widely investigated at the molecular level; however, the exact mechanisms driving the differentiation process are still not fully understood. Especially, limited information persists concerning the role of the PKC superfamily during the cardiomyocyte differentiation process.ObjectiveHere, we employed an established model for differentiating P19CL6 cells into cardiomyocytes presenting specific beating clusters in vitro, and investigated the role of PKC isotypes and cardiomyocyte differentiation capacity.ResultsP19CL6 cells were fully differentiated into beating cardiomyocytes after 1% DMSO treatment for 16 days. Cardiomyocyte-specific genes including GATA4 and Tbx5 were significantly increased during the differentiation period. PKC isotype expressions, especially alpha and delta, were significantly increased during cardiomyocyte differentiation of P19CL6 cells. To confirm the specific function of PKC isotypes (PKC alpha and delta), an overexpression system was conducted. FACS results indicated that PKC alpha-overexpressed P19CL6 cells had fewer apoptotic cells compared to PKC delta-overexpressed cells and pcDNA3.1(+) vector expressed cells. PKC alpha and delta overexpressed P19CL6 cells showed significant Tbx5 protein expression compared to pcDNA3.1(+) vector expressed cells.ConclusionPKC isotype alpha and delta can regulate the cardiomyocyte differentiation potential of P19CL6 cells. Especially, PKC alpha showed a more effective role in cardiomyocyte differentiation with increased Tbx5 protein expression and decreased apoptotic cell populations compared to PKC delta.