Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 β3 chain and induces cell migration

Thirupandiyur S. Udayakumar, Man Ling Chen, Elisabeth L. Bair, Dorothea C. Von Bredow, Anne E Cress, Raymond B Nagle, G. Timothy Bowden

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Degradation of the extracellular matrix by proteolytic enzymes is a central aspect of physiological and pathologic tissue-remodeling processes such as trophoblastic implantation, wound healing, and tumor invasion. We have hypothesized that prostate adenocarcinoma cell invasion through the normal basal lamina is attributable in part to metalloproteinase-induced cleavage of laminin-5 (Ln-5) and enhanced motility of the cancer cells. We studied the role of membrane type-1-matrix metalloproteinase (MT1-MMP) expressed on the surface of prostate tumor cells in cleaving Ln-5 and enhancing the migration of prostate tumor cells. We also determined the nature of the MT1-MMP cleavage of human Ln-5 and how this altered Ln-5 changes the migration of prostate carcinoma cells. We found that human MT1-MMP cleaves purified human Ln-5 to an 80-kDa fragment. Mass spectrometry analyses of the 80-kDa cleaved product by trypsin and chymotrypsin gave 14 and 9 different peptide sequences, respectively, that were identical to the expected amino acid sequence of the Ln-5-β3 chain. The recovered peptides represent 14.4% (trypsin) and 10.3% (chymotrypsin) of Ln-5-β3 chain by amino acid count. Both trypsin and chymotrypsin digestion of MT1-MMP-cleaved product of Ln-5 did not show any other peptides that were identical to the other chains of Ln-5. Using a linear migration assay we found that the Ln-5 cleaved by MT1-MMP enhanced the migration of DU-145 prostate carcinoma cells by 2-fold compared with uncleaved Ln-5. The use of blocked antisense MT1-MMP oligonucleotides inhibited the migration of DU-145 cells on Ln-5. We also found that the prostate carcinoma cells expressing high levels of MT1-MMP, such as PC3N and PPC, demonstrated enhanced migration on human Ln-5-coated substrate, and this migration was inhibited using blocked antisense MT1-MMP oligonucleotides. In conclusion, this is a novel and important finding where we have shown that β3-chain is cleaved by MT1-MMP, and this cleavage enhances migration of prostate cancer cells.

Original languageEnglish (US)
Pages (from-to)2292-2299
Number of pages8
JournalCancer Research
Volume63
Issue number9
StatePublished - May 1 2003

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Matrix Metalloproteinase 14
Cell Movement
Prostate
Carcinoma
Oligonucleotides
kalinin
Peptides
Neoplasms
Metalloproteases
Basement Membrane
Wound Healing
Extracellular Matrix

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 β3 chain and induces cell migration. / Udayakumar, Thirupandiyur S.; Chen, Man Ling; Bair, Elisabeth L.; Von Bredow, Dorothea C.; Cress, Anne E; Nagle, Raymond B; Bowden, G. Timothy.

In: Cancer Research, Vol. 63, No. 9, 01.05.2003, p. 2292-2299.

Research output: Contribution to journalArticle

Udayakumar, Thirupandiyur S. ; Chen, Man Ling ; Bair, Elisabeth L. ; Von Bredow, Dorothea C. ; Cress, Anne E ; Nagle, Raymond B ; Bowden, G. Timothy. / Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 β3 chain and induces cell migration. In: Cancer Research. 2003 ; Vol. 63, No. 9. pp. 2292-2299.
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abstract = "Degradation of the extracellular matrix by proteolytic enzymes is a central aspect of physiological and pathologic tissue-remodeling processes such as trophoblastic implantation, wound healing, and tumor invasion. We have hypothesized that prostate adenocarcinoma cell invasion through the normal basal lamina is attributable in part to metalloproteinase-induced cleavage of laminin-5 (Ln-5) and enhanced motility of the cancer cells. We studied the role of membrane type-1-matrix metalloproteinase (MT1-MMP) expressed on the surface of prostate tumor cells in cleaving Ln-5 and enhancing the migration of prostate tumor cells. We also determined the nature of the MT1-MMP cleavage of human Ln-5 and how this altered Ln-5 changes the migration of prostate carcinoma cells. We found that human MT1-MMP cleaves purified human Ln-5 to an 80-kDa fragment. Mass spectrometry analyses of the 80-kDa cleaved product by trypsin and chymotrypsin gave 14 and 9 different peptide sequences, respectively, that were identical to the expected amino acid sequence of the Ln-5-β3 chain. The recovered peptides represent 14.4{\%} (trypsin) and 10.3{\%} (chymotrypsin) of Ln-5-β3 chain by amino acid count. Both trypsin and chymotrypsin digestion of MT1-MMP-cleaved product of Ln-5 did not show any other peptides that were identical to the other chains of Ln-5. Using a linear migration assay we found that the Ln-5 cleaved by MT1-MMP enhanced the migration of DU-145 prostate carcinoma cells by 2-fold compared with uncleaved Ln-5. The use of blocked antisense MT1-MMP oligonucleotides inhibited the migration of DU-145 cells on Ln-5. We also found that the prostate carcinoma cells expressing high levels of MT1-MMP, such as PC3N and PPC, demonstrated enhanced migration on human Ln-5-coated substrate, and this migration was inhibited using blocked antisense MT1-MMP oligonucleotides. In conclusion, this is a novel and important finding where we have shown that β3-chain is cleaved by MT1-MMP, and this cleavage enhances migration of prostate cancer cells.",
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AU - Bair, Elisabeth L.

AU - Von Bredow, Dorothea C.

AU - Cress, Anne E

AU - Nagle, Raymond B

AU - Bowden, G. Timothy

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