Regulation of docetaxel-induced apoptosis of human melanoma cells by different isoforms of protein kinase C.
Our previous studies showed that docetaxel-induced apoptosis of human melanoma cells was dependent on the activation of the c-jun NH(2)-terminal kinase (JNK) signaling pathway but was inhibited by the extracellular signal-regulated kinase (ERK)-1/2 pathway. However, the mechanisms by which these pathways were modulated by docetaxel were not clear. We report here that docetaxel induces activation of protein kinase C (PKC) signaling differentially through PKCepsilon and PKCdelta isoforms. Activation of PKCepsilon was most marked in docetaxel-resistant cells and paralleled the activation of the ERK1/2 pathway. Inhibition of PKCepsilon by small interfering RNA molecules resulted in down-regulation of phosphorylated ERK1/2 and sensitization of cells to docetaxel-induced apoptosis. Experiments also showed that beta-tubulin class III, a molecular target of docetaxel, coimmunoprecipitated with PKCepsilon and colocalized in confocal microscopic studies. In contrast to PKCepsilon, high levels of activated PKCdelta were associated with activation of the JNK pathway and sensitivity to docetaxel. Activation of PKCdelta seemed to be upstream of JNK because inhibition of PKCdelta by small interfering RNA abrogated activation of the JNK pathway. Although PKCdelta could be activated in resistant cells, downstream activation of JNK and c-Jun did not occur. In summary, these results suggest that the outcome of docetaxel-induced apoptotic events in human melanoma cells depends on their PKC isoform content and signaling responses. PKCepsilon was associated with prosurvival signaling through ERK, whereas PKCdelta was associated with proapoptotic responses through JNK activation.
Mhaidat
Nizar M
NM
Immunology and Oncology Unit, David Maddison Clinical Sciences Building, Newcastle, New South Wales, Australia.
Thorne
Rick F
RF
Zhang
Xu Dong
XD
Hersey
Peter
P
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D045744:Cell Line, Tumor; D000077143:Docetaxel; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D008545:Melanoma / Q000201:enzymology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D051745:Protein Kinase C-delta / Q000378:metabolism*; D051744:Protein Kinase C-epsilon / Q000378:metabolism*; D012878:Skin Neoplasms / Q000201:enzymology*; D043823:Taxoids / Q000494:pharmacology*; D014404:Tubulin / Q000378:metabolism
D000970:Antineoplastic Agents; D007527:Isoenzymes; D043823:Taxoids; D014404:Tubulin; D000077143:Docetaxel; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048670:MAP Kinase Kinase 4