Modulation of type-IV collagenase activity and invasive behavior of metastatic human melanoma (A2058) cells in vitro by monoclonal antibodies to type-IV collagenase.
Monoclonal antibodies (MAbs) against human type-IV collagenase were developed and used for studies on enzyme activity and tumor-cell invasion in vitro. Fifteen MAb clones were generated against the enzyme purified form serum-free culture medium of human melanoma cells (A2058). Five clones affecting the activity of type-IV collagenase were selected for further characterization. All the selected clones could be used for a single-step purification of type-IV collagenase using IgG-Sepharose affinity columns. One of the antibodies activated the enzyme when 3H-proline-labelled type-IV collagen was used as substrate. The activation was dependent on the enzyme antibody ratio. Four clones caused more than 30% inhibition of the activity, maximal inhibition being 50%. Interestingly, the same antibody which activated the enzyme also increased the invasion of A2058 cells through a reconstituted basement membrane in an in vitro invasion assay. The 4 inhibitory antibodies decreased the penetration of A2058 cells through the reconstituted basement membrane. The results strongly support previous findings about the importance of type-IV collagenase in tumor-cell invasion.
Höyhtyä
M
M
Biocenter, University of Oulu, Finland.
Hujanen
E
E
Turpeenniemi-Hujanen
T
T
Thorgeirsson
U
U
Liotta
L A
LA
Tryggvason
K
K
D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology*; D000918:Antibody Specificity; D002460:Cell Line; D002999:Clone Cells / Q000201:enzymology / Q000473:pathology; D003094:Collagen / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007231:Infant, Newborn; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology; D003012:Microbial Collagenase / Q000276:immunology / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000473:pathology
D000911:Antibodies, Monoclonal; D003094:Collagen; D003012:Microbial Collagenase