THE LONG-TERM OBJECTIVE OF THIS PROJECT IS TO ESTABLISH NOVEL RECIPIENT EPITHELIAL CELL SYSTEMS THAT WILL DETECT A WIDE SPECTRUM OF ONCOGENES THROUGH DNA TRANSFECTION.

THE LONG-TERM OBJECTIVE OF THIS PROJECT IS TO ESTABLISH NOVEL RECIPIENT EPITHELIAL CELL SYSTEMS THAT WILL DETECT A WIDE SPECTRUM OF ONCOGENES THROUGH DNA TRANSFECTION. THREE TYPES OF LIVER CELLS REPRESENTING DIFFERENT STAGES OF THE CARCINOGENIC PROCESS WILL BE USED: A NORMAL DIPLOID, AN IMMORTALIZED ANEUPLOID, AND A MUTANT CELL LINE THAT IS TEMPERATURE SENSITIVE FOR TRANSFORMED PHENOTYPE. SPECIFIC AIMS FOR PHASE I INVOLVE: (1) DETERMINATION OF THE TIME COURSE AND DOSE RESPONSE OF DNA UPTAKE IN ORDER TO STANDARD-IZE THE OPTIMAL CONDITIONS OF UPTAKE. THIS WILL BE ACHIEVEDUSING A SENSITIVE ELISA TECHNIQUE THAT ASSAYS FOR BUDR- LABELLED DNA. (2) DETERMINATION OF TRANSFORMATION FREQUEN- CIES INDUCED BY PLASMID DNAS CONTAINING HUMAN HRAS OR V-MYC GENES. CONCLUSIVE RESULTS ON THE BEST TRANSFECTION CONDITIONS (ENHANCEMENT OF DNA UPTAKE BY CELL SURFACE MANIPULATIONS, AND THE EXPRESSION OF ONCOGENE-MEDIATED PHENOTYPE), WILL BE STANDARDIZED. PHASE I RESULTS WILL PROVIDE A STANDARDIZED PROTOCOL FOR FUTURE DNA TRANSFECTION EXPERIMENTS; THESE ARE PLANNED FOR PHASE II USING OTHER CELLTYPES AND ONCOGENES.