Expression of BRCA2 Gene Product in Normal and Breast Cancer Cells in vivo Analysis of its Tumor Suppressor Function.
Abstract
BRCM is a tumor suppressor gene which plays an essential role in cellular proliferation and development. Germ line mutations in BRCM cause familial early onset breast cancer. BRCA2 gene encodes a protein of 3418 amino acids whose function remains largely unclear. The objective of this study is to analyze the tumor suppressor function of BRCA2 gene by manipulating its expression in normal breast cells and breast cancer cells. Towards this goal, cDNAs corresponding to BRCA2 gene were isolated by screening a Jurkat cDNA library. Northern hybridization analysis of total RNA from normal and cancer cell lines indicated an alteration of transcript size in MCF7 and DUl45 cells. Mutational analysis of BRCA2 transcripts from these cell lines is in progress. Full length BRCA2 cDNA was synthesized from a normal breast epithelial cell line, MCFl0A using a combination of RT-PCR and cloning strategies. The entire coding sequence was introduced into two mammalian expression vectors (pBKCMV and ptTAS) in both sense and anti sense orientation. A second anti-sense construct encompassing the 5' untranslated sequences and exons 1-7 was designed and cloned in a tetracycline inducible expression vector, ptTAS. These constructs were characterized by restriction fragment analysis and PCR Expression studies of the corresponding RNAs in transfected normal mammary epithelial cells and breast cancer cell lines is in progress.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1998
- Accession Number
- ADA356096
Entities
People
- Jacob K. Arakkal-nellissery
Organizations
- Temple University