Amplified Genes in Breast Cancer: Molecular Targets for Investigation and Therapy.
Annual rept. 1 Sep 94-31 Aug 95,
SALK INST FOR BIOLOGICAL STUDIES LA JOLLA CA
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The primary objective has been development of a rapid method for purifying extrachromosomally amplified genes DMs from tumor cell lines without relying on cytogenetics. DMs were purified by isolating micronuclei induced by treatment with hydroxyurea. Micronuclei were dissociated from nuclei by lysis under conditions that disaggregated intermediate filaments. Rate zonal and density centrifugation steps completely separated micronuclei from nuclei, enabling preparation of probes for fluorescence in situ hybridization to metaphase chromosomes of normal peripheral blood lymphocytes to identify the genomic regions from which the DMs were generated. Four cell lines derived from different types of tumors with DM contents spanning the range typically reported in human biopsy material e.g., 2-50 per nucleus were analyzed.
- Medicine and Medical Research