Degradation of type IV collagen by neoplastic human skin fibroblasts
An assay for the degradation of type IV (basement membrane) collagen was developed as a biochemical marker for neoplastic cells from chemically transformed human skin fibroblasts. Type IV collagen was isolated from basement membrane of Syrian hamster lung and type I collagen was isolated from rat tails; the collagens were radioactively labelled by reductive alkylation. The abilities of normal (KD) and chemically transformed (Hut-11A) human skin fibroblasts to degrade the collagens were studied. A cell-associated assay was performed by growing either normal or transformed cells in the presence of radioactively labelled type IV collagen and measuring the released soluble peptides in the medium. This assay also demonstrated that KD cells failed to synthesize an activity capable of degrading type IV collagen whereas Hut-11A cells degraded type IV collagen in a linear manner for up to 4 h. Human serum at very low concentrations, EDTA and L-cysteine inhibited the enzyme activity, whereas protease inhibitors like phenylmethyl sulfonyl fluoride, N-ethyl maleimide or soybean trypsin inhibitor did not inhibit the enzyme from Hut-11A cells. These results suggest that the ability to degrade specifically type IV collagen may be an important marker for neoplastic human fibroblasts and supports a role for this collagenase in tumor cell invasion.
- Research Organization:
- National Institute of Environmental Health Sciences, Research Triangle Park, NC
- OSTI ID:
- 5613091
- Journal Information:
- Carcinogenesis (N.Y.); (United States), Vol. 6:2
- Country of Publication:
- United States
- Language:
- English
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COLLAGEN
LABELLING
PROTEOLYSIS
TISSUE DISTRIBUTION
PEPTIDE HYDROLASES
CELL CULTURES
ELECTROPHORESIS
FIBROBLASTS
MEN
MICE
RATS
TRACER TECHNIQUES
TUMOR CELLS
ANIMAL CELLS
ANIMALS
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
DECOMPOSITION
DISTRIBUTION
ENZYMES
HYDROLASES
ISOTOPE APPLICATIONS
MALES
MAMMALS
MAN
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
RODENTS
SCLEROPROTEINS
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
550301 - Cytology- Tracer Techniques