Immunoassays of the metalloproteinase (MMP-2) and tissue inhibitor of metalloproteinase (TIMP 1 and 2) levels in noninvasive and metastatic PC-3 clones: effects of taxol.
We have selected and subcloned bone metastatic (PC-3 ML) and noninvasive, nonmetastatic (3 x N.I.) lines from human prostatic PC-3 parent cells. In this paper, we have compared relative levels of MMP-2 and TIMP-1 and 2 in PC-3 tumors grown in severe combined immunodeficient or SCID mice. Dot blots with polyclonal antibodies specific for MMP-2, TIMP-1 and TIMP-2 revealed that the MMP-2 levels were high in subcutaneously grown PC-3 ML clones but low in 3 x N.I. clones at days 20 and 40. The TIMP-1 levels were inversely proportional to MMP-2 in the two types of clones, respectively. The TIMP-2 levels were similar in both clones at days 20 and 40. Gelatin zymograms confirmed that PC-3 ML tumors contained MMP-2 (and not MMP-9) subcutaneously or in bone metastases in SCID mice. Slot blots of PC-3 ML bone tumors comparing MMP-2 and TIMP-1 levels showed at days 10, 15, 17, 19, 21, and 25 that the ratio of MMP-2 to TIMP-1 increased, especially at about day 21, when extensive secondary metastases to the peritoneal cavity occurred. The levels of TIMP-2 remained constant. Quantitative ELISAs confirmed the blotting data and showed that taxol blocked MMP-2 but not TIMP-1 production in these advanced tumors. We conclude that highly metastatic PC-3 ML variants contained relatively high levels of MMP-2 and low amounts of TIMP-1.
Department of Pathology, Medical College of Pennsylvania, Philadelphia 19129.
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D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000556:secondary; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D018093:Gelatinases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D006801:Humans; D007118:Immunoassay; D015151:Immunoblotting; D008297:Male; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D051379:Mice; D016513:Mice, SCID; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D017239:Paclitaxel / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D019716:Tissue Inhibitor of Metalloproteinase-2; D019714:Tissue Inhibitor of Metalloproteinases; D014407:Tumor Cells, Cultured
D006023:Glycoproteins; D011506:Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D017239:Paclitaxel