Identification and Localization of Genes Which Restore Senescence in Breast Cancer Cells.
Annual rept. 30 Aug 96-29 Aug 99
TEMPLE UNIV PHILADELPHIA PA
Pagination or Media Count:
Using a combination of positional and functional cloning, we have mapped SEN 16 to a lO0xb region at 16q24.3. Intact, normal human chromosome 16, when transferred into immortal human MCF SKBR-3 and rat LA7 NMU mammary tumor cells by microcell mediated chromosome transfer MMCT restored senescence to these cells. Moreover, transfer of a fragment of chromosome 16 containing 16q23-qter also induced senescence to these tumor cells. Continuous culture of the senescent cells in selection media gave rise to revertant cells which have parental cell growth phenotype and have lost the region of the introduced chromosome that harbors the senescence gene. Analysis of these revertant clones with previously mapped chromosome 16-specific markers placed the senescence gene within a 3-7 cm region at 16q24.3. Three overlapping Yeast-Artificial-Chromosome YAC clones spanning this region were obtained. Transfer of one of these YACs, 792El 360 Kb, restored senescence in MCF and LA7 cells while another YAC containing a portion of chromosome 6 did not alter the growth characteristics of these cells. To further map SENl6, a partial Bacterial-Artificial-Chromosome BAC contig was created spanning the YAC 792E1. Experiments are in progress to identify a BAC that harbors SENl 6 and to further clone and characterize, this gene.
- Anatomy and Physiology
- Medicine and Medical Research