QUANTIFICATION OF ONCOGENE PRODUCTS IN CELLS AND TISSUE

THE OBJECTIVE OF THIS PROJECT IS TO CONCEPTUALIZE AN ASSAY METHOD TO QUANTITATE ONCOGENE PRODUCTS IN CELLS AND TISSUES. IN THE PHASE I EFFORTS, THE PROTEIN ENCODED BY THEHER2 ONCOGENE, P185(HER2), WILL BE USED AS A SPECIFIC EXAMPLE BECAUSE IT HAS BEEN STRONGLY IMPLICATED IN THE BIOLOGICAL BEHAVIOR OF BREAST CANCER AND MAY HAVE PROGNOSTICSIGNIFICANCE. THE TECHNIQUES USED WILL BE THOSE OF HIGH-RESOLUTION DIGITALIMAGE MICROSCOPY AND IMMUNOHISTOCHEMICAL LOCALIZATION. SINCE IMMUNOHISTOCHEMISTRY PERMITS ANALYSIS OF INDIVIDUAL CELLS, VERY SMALL TISSUE SAMPLES CAN BE USED, ENABLING THIS ASSAY TO BE PERFORMED ON SMALL LESIONS, BIOPSIES, OR CYTOLOGICAL SMEARS OF FINE-NEEDLE ASPIRATES OF BREAST TUMORS. HOWEVER, THE DISADVANTAGE OF IMMUNOHISTOCHEMICAL TECHNIQUES IS THE DIFFICULTY THEY PRESENT IN QUANTITATING EXPRESSION LEVELS IN AN OBJECTIVE MANNER. THIS RESEARCH IS AIMED AT IMPROVING THIS LIMITATION. THE PROPOSED ASSAY METHOD, RELYING ON DIGITAL IMAGE PROCESSING, WILL BE A QUANTITATIVE ASSAY FOR ONCOGENE EXPRESSION IN INDIVIDUAL CELLS THAT CAN BE PERFORMED EASILY AND QUICKLY ON THE MALIGNANT CELLS OF A SPECIMEN SAMPLE.