Matrix metalloproteinase inhibition suppresses MMP-2 activity and activation of PANC-1 cells in vitro.
BACKGROUND
We have shown previously that the metalloproteinase inhibitor, BB-94, prolongs survival and attenuates MMP-2 activity in a murine model of pancreatic cancer. The purpose of this study was to determine the effect of BB-94 on the activity and activation of MMP-2 by PANC-1 cells in vitro.


METHODS
The poorly differentiated pancreatic cancer cell line PANC-1 was stimulated in vitro with the phorbol ester PMA (20 nM) and grown in the presence of increasing doses of BB-94 (0, 40, 200, and 400 ng/ml) for 24 h. Activation of MMP-2 was determined by gel zymography. In a separate experiment detailing the effects of BB-94 on MMP-2 activity, PANC cells were stimulated for 24 h with PMA and run out on four separate zymograms, each incubated in the previously noted concentrations of BB-94. Using densitometry, band intensity on all gels was determined and compared for each concentration of BB-94. The Matrigel assay was used to determine BB-94's effect on the invasive potential of PANC cells at the previously studied concentrations. The presence of MT-MMP (a putative component of MMP-2 activation) was confirmed using Western blot in each group.


RESULTS
BB-94 inhibited the conversion of latent to active MMP-2 in a dose-dependent fashion. BB-94 also inhibited the activity of MMP-2 when run out on gel zymograms incubated with increasing concentrations of BB-94. Decreased activity and activation of MMP-2 by BB-94 were manifested by a significant reduction in the invasive potential of PANC as determined by the Matrigel assay. MT-MMP was universally present in each study group.


CONCLUSIONS
The previously described salutary effects of MMP blockade in mice implanted with pancreatic cancer can be explained in vitro by a dose-dependent diminution of MMP-2 activity and activation in PANC cells exposed to BB-94.
Zervos
E E
EE
Department of Surgery, University of South Florida College of Medicine, Tampa, Florida, 33601, USA.
Shafii
A E
AE
Haq
M
M
Rosemurgy
A S
AS
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D018093:Gelatinases / Q000037:antagonists & inhibitors*; D020778:Matrix Metalloproteinase 2; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D009361:Neoplasm Invasiveness / Q000473:pathology; D009994:Osmolar Concentration; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000473:pathology; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D013876:Thiophenes / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D011480:Protease Inhibitors; D013876:Thiophenes; D010649:Phenylalanine; C080985:batimastat; D018093:Gelatinases; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2