Rapid tyrosine phosphorylation of HS1 in the response of mouse lymphoma L5178Y-R cells to photodynamic treatment sensitized by the phthalocyanine Pc 4.
The ability of photodynamic treatment (PDT) with the phthalocyanine Pc 4 to activate cellular signal transduction pathways in murine lymphoma L5178Y-R cells has been assessed by observing increases in protein tyrosine phosphorylation at early times post-PDT. Western blot analysis with an anti-phosphotyrosine antibody revealed a dramatic increase in phosphorylation of two major protein bands of Mr approximately 80,000 and approximately 55,000 in response to PDT. The increase was PDT dose-dependent, occurred as early as 20 s after initiation of light exposure of Pc 4-preloaded cells and was amplified by the presence of the protein tyrosine phosphatase inhibitor, sodium orthovanadate (NaVO4). By immunoprecipitation, one of the Mr approximately 80,000 phosphorylated proteins has been identified as HS1, a substrate of nonreceptor-type protein tyrosine kinases. Although vanadate greatly enhanced the level and extent of PDT-induced phosphorylation, it had no influence on overall photocytotoxicity or on the rate of apoptotic DNA fragmentation. Genistein, an inhibitor of protein tyrosine kinases, diminished tyrosine phosphorylation of the Mr approximately 80,000 and other proteins and dramatically potentiated cell killing induced by PDT but did not significantly affect PDT-induced apoptosis. The results suggest that PDT rapidly activates a membrane-associated src family kinase(s) in L5178Y-R cells, one substrate of which is HS1, and that protein tyrosine phosphorylation is part of a stress response, protecting a portion of the cells from the lethal effects of PDT but not altering the mechanism by which they die.
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Division of Radiation Biology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
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Oleinick
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D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001798:Blood Proteins / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D016923:Cell Death / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein; D007211:Indoles / Q000633:toxicity*; D007529:Isoflavones / Q000494:pharmacology; D007940:Leukemia L5178; D008027:Light; D051379:Mice; D008970:Molecular Weight; D010766:Phosphorylation; D019000:Phosphotyrosine; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000633:toxicity*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology
D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins; D004791:Enzyme Inhibitors; C061909:HCLS1 protein, human; D007211:Indoles; D007529:Isoflavones; D017319:Photosensitizing Agents; C107448:phthalocyanine Pc 4; D019000:Phosphotyrosine; D014638:Vanadates; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases