<p>Doxorubicin (DOX)-induced cardiotoxicity (DIC) is known to be associated with reduction of cardiac protein kinase C epsilon (PKC-ε). PKC-ε promotes cell survival and protects hearts against various stresses. However, it is unclear whether or not the reduction in cardiac PKC-ε expression plays a causal role in DIC and in particular the potential underlying mechanism whereby PKC-ε may protect against DIC. C57BL/6 mice (8–10-week-old) were either treated with DOX administered intraperitoneally for a duration of 4&#xa0;weeks to produce cardiotoxicity, or untreated in which mice received the same volume of saline. In vitro, neonatal rat ventricle cardiomyocytes were exposed to DOX for 24&#xa0;h in the absence or presence of adenovirus overexpressing PKC-ε. Cardiomyocytes in a subgroup were treated with sirtuin-1 (SIRT1) selective inhibitor Ex527. Four weeks after DOX, cardiac contractile function was decreased concomitant with increased serum CK-MB and LDH levels as well as increases in Bax-to-Bcl-2 ratio and Cleaved Caspase 3 proteins expression, while PKC-ε and Sirt1 protein expressions were significantly decreased. In vitro, DOX reduced cardiomyocyte PKC-ε and SIRT1 protein expression, decreased cardiomyocyte viability, and increased LDH release with concomitant increases in oxidative stress and apoptosis. These changes were attenuated by overexpression of PKC-ε. IP study showed that PKC-ε could directly or indirectly bind SIRT1 in cardiomyocytes, and the protect effects of PKC-ε were further canceled by SIRT1 inhibition. In conclusion, activating SIRT1 may represent a major mechanism whereby PKC-ε protects the heart against DOX-induced cell apoptosis and oxidative stress.</p> Graphical Abstract <p>Dox induces cardiotoxicity via inhibiting PKC-epsilon/Sirt1 signaling which can be reversed by PKC-epsilon overexpression.</p> <p></p>