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

Oncol Res. 1994;6(4-5):195-201.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Antibody Specificity
  • Blotting, Western
  • Bone Neoplasms / drug therapy*
  • Bone Neoplasms / metabolism*
  • Bone Neoplasms / secondary
  • Cell Division / drug effects
  • Clone Cells
  • Enzyme-Linked Immunosorbent Assay
  • Gelatinases / analysis
  • Gelatinases / antagonists & inhibitors
  • Gelatinases / biosynthesis*
  • Glycoproteins / analysis
  • Glycoproteins / biosynthesis*
  • Humans
  • Immunoassay
  • Immunoblotting
  • Male
  • Matrix Metalloproteinase 2
  • Metalloendopeptidases / analysis
  • Metalloendopeptidases / antagonists & inhibitors
  • Metalloendopeptidases / biosynthesis*
  • Mice
  • Mice, SCID
  • Neoplasm Invasiveness
  • Neoplasm Transplantation
  • Paclitaxel / pharmacology*
  • Prostatic Neoplasms / drug therapy*
  • Prostatic Neoplasms / metabolism*
  • Prostatic Neoplasms / pathology
  • Protein Biosynthesis*
  • Proteins / analysis
  • Tissue Inhibitor of Metalloproteinase-2
  • Tissue Inhibitor of Metalloproteinases
  • Tumor Cells, Cultured

Substances

  • Glycoproteins
  • Proteins
  • Tissue Inhibitor of Metalloproteinases
  • Tissue Inhibitor of Metalloproteinase-2
  • Gelatinases
  • Metalloendopeptidases
  • Matrix Metalloproteinase 2
  • Paclitaxel