Accession Number : ADA298514

Title :   Genetic Construction and Molecular Characterization of Breast Cancer Precursor Cells.

Descriptive Note : Annual rept. 31 May 94-30 May 95,

Corporate Author : UTAH UNIV SALT LAKE CITY

Personal Author(s) : Spancake, Kimberly M.

PDF Url : ADA298514

Report Date : 30 JUN 1995

Pagination or Media Count : 12

Abstract : Mutations in genes involved in cell-cycle and growth regulation, such as the retinoblastoma gene (RB), are expected to have a significant role in the early stages of breast cancer. RB-deficient cells are being created by two distinct methods. First, targeting vectors, currently being constructed, will be transfected into human mammary epithelial cell lines (HMEC) in order to knock out both RB allels via homologous recombination. Second, a human papilloma virus (HPV) E6/E7 fusion construct, previously shown to specifically target the retinoblastoma protein (pRb) for degradation, will be transfected into HMEC. The E6/E7 gene fusion will be introduced into the HMEC under the control of a regulatable promoter, allowing for direct regulation of protein expression. Breast cells made deficient for pRb by these two methods will be examined for cellular and molecular alterations, such as changes in morphology and behavior, and their ability to form duct-like structures when grown on or within an extracellular matrix. Additionally, since the normal RB product has been implicated in transcriptional regulation, the effect on mRNA transcription in the RB-deficient breast cells will be assayed by Northern blot analysis and differential display.

Descriptors :   *GENETICS, *CANCER, *MAMMARY GLANDS, DEGRADATION, CELLS, PRECURSORS, GROWTH(GENERAL), PROTEINS, MUTATIONS, DISPLAY SYSTEMS, CONSTRUCTION, GENES.

Subject Categories : Medicine and Medical Research
      Genetic Engineering and Molecular Biology

Distribution Statement : APPROVED FOR PUBLIC RELEASE