Degradation of type IV collagen by neoplastic human skin fibroblasts

Journal Article · Fri Feb 01 04:00:00 EST 1985 · Carcinogenesis (N.Y.); (United States)

DOI:https://doi.org/10.1093/carcin/6.2.173· OSTI ID:5613091

Sheela, S; Barrett, J C

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), Journal Name: Carcinogenesis (N.Y.); (United States) Vol. 6:2; ISSN CRNGD
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
550301 -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
CELL CULTURES
CHEMICAL REACTIONS
COLLAGEN
CONNECTIVE TISSUE CELLS
DECOMPOSITION
DISTRIBUTION
ELECTROPHORESIS
ENZYMES
FIBROBLASTS
HYDROLASES
ISOTOPE APPLICATIONS
LABELLING
MALES
MAMMALS
MAN
MEN
MICE
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
PRIMATES
PROTEINS
PROTEOLYSIS
RATS
RODENTS
SCLEROPROTEINS
SOMATIC CELLS
TISSUE DISTRIBUTION
TRACER TECHNIQUES
TUMOR CELLS
VERTEBRATES