C-kit mutations determine dasatinib mechanism of action in HMC-1 neoplastic mast cells: dasatinib differently regulates PKCδ translocation in HMC-1560 and HMC-1560,816 cell lines

Purpose: The second generation of tyrosine kinase inhibitors is a group of compounds that inhibit c-kit receptor activity and therefore widely used in the treatment of mastocytosis. In this research, the relationship between the mechanism of action of tyrosine kinase inhibitors and protein kinase C is investigated in HMC-1(560) or HMC-1(560,816) cell lines. Results: From all the tyrosine kinase inhibitors tested, nilotinib is the compound that has the highest cytotoxic effect against HMC-1(560) mast cell line, while midostaurin is the most potent in HMC-1(560,816). Moreover, an increase on histamine release is observed after protein kinase C activation either in HMC-1(560) or HMC-1(560,816) cells. Furthermore, dasatinib increases histamine release in both mast cell lines, which could be related with the secondary reactions previously described in dasatinib-treated patients. Dasatinib also induces Ca2+-dependent protein kinase C isoforms translocation from the cytosol to the membrane, whereas protein kinase C delta is translocated from the cytosol to the nucleus in the HMC-1(560,816) cell line, but not in HMC-1(560) cells. Conclusion: Results obtained demonstrate that dasatinib induces an important cytotoxic effect in both HMC-1 cell lines and differently regulates protein kinase Cd in HMC-1(560) and HMC-1(560,816) cells. Finally, our results confirm that PKCd is an essential target for dasatinib.