Collagenases in Breast Cancer Cell-Induced Metastatic Tumor Growth and Progression
Abstract
Since collagenases (-1 and -3) degrade the extracellular bone matrix (ECM) components, collagenase-driven ECM proteolysis may facilitate cancer growth and progression. To test this hypothesis, we proposed to utilize a transgenic mouse model to overexpress collagenase- 1 under the control of the bone specific osteocalcin promoter. The expression plasmid pCMV-collagenase- 1 -V5 was constructed, which contains an eukaryotic promoter (CMV) fragment and a 1.65 kb human collagenase-l cDNA fragment, followed by a V5 epitope tag. The pCMV-collagenase-l-V5 was transiently transfected into COS-7 cells and the expression of collagenase-l-V5 was identified by Western blot using the anti-V5 antibody. The pOC-collagenase- I-V5 was constructed by replacing the CMV promoter sequence from pCMV-collagenase-l-V5 with the rat osteocalcin promoter sequence (OC). The ability of osteocalcin promoter to drive collagenase-l-V5 expression is being carried out by transient transfection assays and Western blot analysis. The 2.6 kb DNA fragment containing the osteocalcin promoter (1 kb) and collagenase- 1-V5 cDNA sequence will be excised from pOC- collagenase- 1-V5 with appropriate enzymes and used for generation of transgenic mice. In addition, the molecular mechanism(s) responsible for transforming growth factor-beta 1 (a crucial molecule in metastatic bone cancer) stimulation of collagenase-3 expression in human breast cancer cells was investigated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 2002
- Accession Number
- ADA407462
Entities
People
- Nagarajan Selvamurugan
Organizations
- Robert Wood Johnson Medical School