--- - |2 The purpose of this study was to evaluate the anti–head and neck squamous cell carcinoma (anti‐HNSCC) cell activity by C6 ceramide and multikinase inhibitor PKC412. Experiments were performed on HNSCC cell lines (SQ20B and SCC‐9) and primary human oral carcinoma cells. Results showed that PKC412 inhibited HNSCC cell proliferation without provoking apoptosis activation. Cotreatment of C6 ceramide significantly augmented PKC412‐induced lethality in HNSCC cells. PKC412 decreased Akt–mammalian target of rapamycin (mTOR) activation in HNSCC cells, facilitated with cotreatment of C6 ceramide. In contrast, exogenous expression of a constitutively active Akt restored Akt–mTOR activation and attenuated lethality by the cotreatment. We propose that Mcl‐1 is a primary resistance factor of PKC412. The cytotoxicity of PKC412 in HNSCC cells was potentiated with Mcl‐1 short hairpin RNA knockdown, but was attenuated with Mcl‐1 overexpression. Intriguingly, C6 ceramide downregulated Mcl‐1 in HNSCC cells. In vivo, PKC412 oral administration inhibited SQ20B xenograft tumor growth in severe combined immunodeficient mice. The antitumor activity of PKC412 was further sensitized with coadministration of liposomal C6 ceramide. Together, we suggest that PKC412 could be further studied as a promising anti‐HNSCC strategy, alone or in combination with C6 ceramide. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9437-9446, December 2018.