Walker 256 cancer cells secrete tissue inhibitor of metalloproteinase-free metalloproteinase-9

Mol Cell Biochem. 2009 Aug;328(1-2):189-99. doi: 10.1007/s11010-009-0089-2. Epub 2009 Mar 29.

Abstract

Walker 256 (W256) cancer cells, developed as ascites in rats, in response to endogenous unidentified stimuli, secrete a gelatinase of apparent molecular mass of 94 kDa, immunologically homologous to the zymogen of matrix metalloproteinase-9 (proMMP-9). After treatment with the activating agent 4-aminophenylmercuric acetate (APMA), affinity-purified W256 gelatinase is converted to a final processed form of 66 kDa in a similar fashion to TIMP-free human proMMP-9. It is demonstrated that although being capable of binding TIMP-1, W256 proMMP-9 is secreted from W256 cells in TIMP-free forms (monomers or oligomers). Moreover, using biochemical and immunological methods, it is established that the W256 cells do not express or secrete TIMP-1 protein, although RT-PCR analysis indicated low-level TIMP-1 mRNA expression. W256 cancer cells displayed high metastatic ability in rats that may be attributed in part to secretion of TIMP-free proMMP-9.

MeSH terms

  • Animals
  • Carcinoma 256, Walker / metabolism*
  • Cell Line, Tumor
  • Gelatinases
  • Humans
  • Matrix Metalloproteinase 9 / metabolism*
  • Neoplasm Metastasis
  • Phenylmercuric Acetate / analogs & derivatives
  • Phenylmercuric Acetate / pharmacology
  • RNA, Messenger / analysis
  • Rats
  • Tissue Inhibitor of Metalloproteinase-1 / genetics
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism*

Substances

  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinase-1
  • 4-aminophenylmercuriacetate
  • Gelatinases
  • Matrix Metalloproteinase 9
  • Phenylmercuric Acetate