Identification of Tumor Suppressor Genes in Breast Cancer by Insertional Mutagenesis and Functional Inactivation
Abstract
The development and progression of cancer result from multiple genetic changes accumulated in the cells. The identification of tumor suppressor genes inactivated and proto-oncogenes activated in mammary epithelial cells is essential to understand the genetic basis of breast cancer and is a prerequisite for development of strategies for prevention, diagnosis, and treatment. In breast cancer, loss of heterozygosity (LOH) was detected frequently on chromosome 17 and other chromosomes, suggesting unrecognized tumor suppressor genes. We are applying the novel retroviral-tagging strategy to identify the genes using chromosome 17-suppressed (independent of p53 and BRCAl) breast cancer cell lines. In contrast to the parental tumorigenic cell line CAL51, the suppressed sublines CAL/17-3 and CAL/17-5 display retard growth in flasks, no growth in soft-agar culture and athymic nude mice. In this annual report, we present preparation of biological materials including cell culture, isolation of DNA and RNA, construction of cDNA library and packaging retrovirus particles. In order to provide antisense and mutant proteins to inhibit activities of tumor suppressor genes, poly-(A)+RNA was isolated from the suppressed subline CAL17-3 and used to construct the library. We are now in the process to package cDNA library into retrovirus particles for transduction.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1998
- Accession Number
- ADA366888
Entities
People
- Yan A. Su
Organizations
- National Institutes of Health