A functional proteomics screen of proteases in colorectal carcinoma.

J. H. McKerrow, V. Bhargava, E. Hansell, S. Huling, T. Kuwahara, M. Matley, Lisa Coussens, R. Warren

Research output: Contribution to journalArticle

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Abstract

BACKGROUND: Proteases facilitate several steps in cancer progression. To identify proteases most suitable for drug targeting, actual enzyme activity and not messenger RNA levels or immunoassay of protein is the ideal assay readout. MATERIALS AND METHODS: An automated microtiter plate assay format was modified to allow detection of all four major classes of proteases in tissue samples. Fifteen sets of colorectal carcinoma biopsies representing primary tumor, adjacent normal colon, and liver metastases were screened for protease activity. RESULTS: The major proteases detected were matrix metalloproteases (MMP9, MMP2, and MMP1), cathepsin B, cathepsin D, and the mast cell serine proteases, tryptase and chymase. Matrix metalloproteases were expressed at higher levels in the primary tumor than in adjacent normal tissue. The mast cell proteases, in contrast, were at very high levels in adjacent normal tissue, and not detectable in the metastases. Cathepsin B activity was significantly higher in the primary tumor, and highest in the metastases. The major proteases detected by activity assays were then localized in biopsy sections by immunohistochemistry. Mast cell proteases were abundant in adjacent normal tissue, because of infiltration of the lamina propria by mast cells. Matrix metalloproteases were localized to the tumor cells themselves; whereas, cathepsin B was predominantly expressed by macrophages at the leading edge of invading tumors. Although only low levels of urinary plasminogen activator were detected by direct enzyme assay, immunohistochemistry showed abundant protein within the tumor. CONCLUSIONS: This analysis, surveying all major classes of proteases by assays of activity rather than immunolocalization or in situ hybridization alone, serves to identify proteases whose activity is not completely balanced by endogenous inhibitors and which may be essential for tumor progression. These proteases are logical targets for initial efforts to produce low molecular weight protease inhibitors as potential chemotherapy.

Original languageEnglish (US)
Pages (from-to)450-460
Number of pages11
JournalMolecular medicine (Cambridge, Mass.)
Volume6
Issue number5
StatePublished - May 2000
Externally publishedYes

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Proteomics
Colorectal Neoplasms
Peptide Hydrolases
Mast Cells
Cathepsin B
Neoplasms
Metalloproteases
Neoplasm Metastasis
Immunohistochemistry
Chymases
Biopsy
Tryptases
Cathepsin D
Urokinase-Type Plasminogen Activator
Enzyme Assays
Serine Proteases
Drug Delivery Systems
Protease Inhibitors
Immunoassay
In Situ Hybridization

ASJC Scopus subject areas

  • Genetics

Cite this

McKerrow, J. H., Bhargava, V., Hansell, E., Huling, S., Kuwahara, T., Matley, M., ... Warren, R. (2000). A functional proteomics screen of proteases in colorectal carcinoma. Molecular medicine (Cambridge, Mass.), 6(5), 450-460.

A functional proteomics screen of proteases in colorectal carcinoma. / McKerrow, J. H.; Bhargava, V.; Hansell, E.; Huling, S.; Kuwahara, T.; Matley, M.; Coussens, Lisa; Warren, R.

In: Molecular medicine (Cambridge, Mass.), Vol. 6, No. 5, 05.2000, p. 450-460.

Research output: Contribution to journalArticle

McKerrow, JH, Bhargava, V, Hansell, E, Huling, S, Kuwahara, T, Matley, M, Coussens, L & Warren, R 2000, 'A functional proteomics screen of proteases in colorectal carcinoma.', Molecular medicine (Cambridge, Mass.), vol. 6, no. 5, pp. 450-460.
McKerrow JH, Bhargava V, Hansell E, Huling S, Kuwahara T, Matley M et al. A functional proteomics screen of proteases in colorectal carcinoma. Molecular medicine (Cambridge, Mass.). 2000 May;6(5):450-460.
McKerrow, J. H. ; Bhargava, V. ; Hansell, E. ; Huling, S. ; Kuwahara, T. ; Matley, M. ; Coussens, Lisa ; Warren, R. / A functional proteomics screen of proteases in colorectal carcinoma. In: Molecular medicine (Cambridge, Mass.). 2000 ; Vol. 6, No. 5. pp. 450-460.
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