Cell-cell and cell-collagen interactions influence gelatinase production by human breast-carcinoma cell line 8701-BC.
We previously produced evidence that the human mammary-carcinoma cell line 8701-BC expresses several metalloproteinases (MMP-1, -2, -9, and -10) and their tissue inhibitors). In order to obtain a better understanding of the environmental control over gelatinolytic activities, we have tested the enzyme production of 8701-BC cells, at time intervals after plating on different collagen substrates, i.e., types I, III, IV, V and OF/LB, used as films in culture dishes. Proteinase activities, released in the conditioned culture media, were tested by zymography on SDS-PAGE, and by quantificative analyses, using 14C carboxymethylated transferrin as substrate in a liquid incubation medium. Enzymatic activities varied with time and were inversely related to cell densities, with minimum values at cell confluence. The enzymatic activity was positively supported by collagen substrates, with a maximal increase in activity when OF/LB collagen was used. In addition to the known MMPs, we found a proteinase with an M(r) of about 20 kDa, which displayed higher activity at 48 hr after cell plating and gradually decreased with cell increment. In contrast to the other MMPs, this proteinase is inhibited by soybean trypsin inhibitor, but it does not display a complete identity with trypsin, since it does not digest casein and is not inhibited by other serine proteinase inhibitors.
Minafra
S
S
Institute of Histology and Embryology, University of Palermo, Italy.
Giambelluca
C
C
Andriolo
M
M
Pucci-Minafra
I
I
D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology*; D002450:Cell Communication / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D003094:Collagen / Q000494:pharmacology*; D003470:Culture Media; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism; D018093:Gelatinases / Q000096:biosynthesis*; D006801:Humans; D008970:Molecular Weight; D014407:Tumor Cells, Cultured
D003470:Culture Media; D003094:Collagen; D010450:Endopeptidases; D018093:Gelatinases