Use of Active State Specific Antibodies to Shc for Assessing Breast-Cancer Prognosis
Annual rept. 1 Jul 1999-30 Jun 2000
ROGER WILLIAMS MEDICAL CENTER PROVIDENCE RI
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There is an urgent need for better prognostic indicators in breast cancer. We have found that I the p52p46 Shc signaling protein is constitutively tyrosine-phosphorylated in most breast cancer cell lines, but not in non-tumorigenic breast epithelial cell lines ii expression of a 66-kDa isoform of Shc is greatly reduced in most breast cancer cell lines. The current study, then, ultimately seeks to answer the question, Are the cellular levels of PTyr-Shc and p66 Shc of prognostic value in breast cancer To be able to address this question, we have developed and characterized antibodies that specifically recognize, respectively, either activated Shc phospho-specific antibodies to the Shc tyrosine3l7 phosphorylation site, or to the p66-Shc-specific CH2 domain. These antibodies not only will specifically immunoprecipitate their respective antigens, but will specifically detect these antigens in formalin-fixed tissue culture cells. These antibodies will be used to immunohistochemically detect and quantify the levels of these proteins in patients tumors, and the levels will be correlated retrospectively with the aggressiveness of patients disease.
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