The tyrosine kinase inhibitors methyl 2,5-dihydroxycinnamate and genistein reduce thrombin-evoked tyrosine phosphorylation and Ca2+ entry in human platelets.

Platelet activation is associated with the phosphorylation of a number of platelet proteins at tyrosine residues. The significance of this is unknown. Here we have investigated the effects of two tyrosine kinase inhibitors, methyl 2,5-dihydroxycinnamate and genistein, on thrombin-evoked protein tyrosine phosphorylation and Ca2+ signal generation in fura-2-loaded human platelets. Both compounds inhibited thrombin-evoked tyrosine phosphorylation and reduced the elevation of [Ca2+]i in the presence, but not the absence, of external Ca2+. This suggested a selective inhibition of thrombin-evoked Ca2+ entry but not release from internal stores. Both compounds also reduced thrombin-evoked Mn2+ entry. In contrast, selective blockade of protein kinase C with Ro 31/8220-002 potentiated the thrombin-evoked Ca2+ signal. These data are compatible with a role for protein tyrosine phosphorylation contributing to thrombin-evoked Ca2+ entry in human platelets.