Suppressor Genes in Breast Cancer.
Abstract
Several tumor suppressor genes (TSGs) have been cloned and found to be mutated in a variety of cancers, including breast cancer. However, few breast cancer-specific TSGs are known. The purposes of this proposal are to: (1) clone novel TSGs specific to human breast cancer; (2) examine the alteration of these TSGs in primary breast tumors; and (3) identify their characteristics, regulation and function. We are utilizing the tetracycline (tet) regulable system. We have constructed a cDNA library from normal human breast epithelia and cloned this cDNA library into a vector that is negatively regulated by tet repressor (tetR) and simultaneously expresses enhanced green fluorescence. These vectors were then co-transfected into LCC6, 231, and MCF-7 cells that have the capability to express tetR. Upon withdrawal of tet, the repressed expression of the cDNA of interest is released, and the cDNA is expressed. Using a novel dye that was retained in nonproliferating cells, we were able to identify growth inhibited clones which were then sorted by Flow Cytometry. This functional screen has provided the basis for identifying TSGs which are expressed in the growth inhibited cells. Using PCR, we have obtained the insert sequences. We will now characterize these genes and begin to assess their function and expression in primary breast carcinomas.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1999
- Accession Number
- ADA377185
Entities
People
- Le-ping Pu
- Robert Clarke
Organizations
- Georgetown University