Differences between scirrhous and non-scirrhous human gastric carcinomas from the aspect of proMMP-2 activation regulated by TIMP-3

Takeyoshi Yokoyama, Hiroyuki Nakamura, Yoshihide Otani, Tetsuro Kubota, Noboru Fujimoto, Motoharu Seiki, Masaki Kitajima, Yasunori Okada

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Abstract

Gastric carcinomas can be classified into scirrhous carcinomas (SC), i.e. 'linitis plastica' or Borrmann 4 gastric cancer, and non-scirrhous carcinomas (NSC). SC are characterized by diffuse invasive growth patterns with marked fibrosis, frequent peritoneal dissemination and lymph-node metastases and poor prognosis, while NSC show medullary growth patterns and common hematogenous metastases. To study the differences in local expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) between SC and NSC, we examined the expression of MMPs and TIMPs in human gastric carcinoma tissues by several methods including sandwich-enzyme immunoassay systems, gelatin zymography, reverse transcriptase-polymerase chain reaction (RT-PCR), real-time quantitative PCR, immunoblotting, immunohistochemistry and in situ zymography. Of the seven MMPs and two TIMPs tested, only proMMP-2 levels were remarkably higher in SC than in NSC (P<0.01), and proMMP-2 activation ratio was significantly lower in SC than in NSC (P<0.05). TIMP-3 mRNA levels were remarkably about 2-fold higher in SC than in NSC tissues (P<0.01). TIMP-3 production in SC was confirmed by immunoblotting and TIMP-3 was immunolocalized to stromal fibroblasts in SC. TIMP-3 mRNA levels inversely correlated with proMMP-2 activation ratios, although the expression levels of MT1-MMP and MT2-MMP were not different in SC and NSC. By in situ zymography, gelatinolytic activity appeared to be weaker in SC than in NSC. All these data suggest that proMMP-2 activation is down-regulated by TIMP-3 expressed in scirrhous gastric carcinomas. Our findings may explain the differences in clinical behaviors of SC and NSC. Abbreviations: EIA - sandwich enzyme immunoassay; GAPDH - glyceraldehyde-3-phosphate dehydrogenase. MMP - matrix metalloproteinase; MT-MMP - membrane-type MMP; N - non-carcinoma; NSC - non-scirrhous carcinomas; RT-PCR - reverse transcriptase-polymerase chain reaction; SC - scirrhous carcinomas; SDS-PAGE - sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TIMP - tissue inhibitor of metalloproteinases.

Original languageEnglish
Pages (from-to)223-233
Number of pages11
JournalClinical and Experimental Metastasis
Volume21
Issue number3
DOIs
Publication statusPublished - 2004

Fingerprint

Scirrhous Adenocarcinoma
Tissue Inhibitor of Metalloproteinase-3
Stomach
Carcinoma
Tissue Inhibitor of Metalloproteinases
Matrix Metalloproteinases
Reverse Transcriptase Polymerase Chain Reaction
Matrix Metalloproteinase Inhibitors
progelatinase
Immunoenzyme Techniques
Immunoblotting
Matrix Metalloproteinase 15
Polyacrylamide Gel Electrophoresis
Membrane-Associated Matrix Metalloproteinases
Linitis Plastica
Peritoneal Fibrosis
Neoplasm Metastasis
Matrix Metalloproteinase 14
Messenger RNA
Medullary Carcinoma

Keywords

  • MMP
  • MT1-MMP
  • ProMMP-2 activation
  • scirrhous gastric carcinoma
  • TIMP-3

ASJC Scopus subject areas

  • Cancer Research

Cite this

Differences between scirrhous and non-scirrhous human gastric carcinomas from the aspect of proMMP-2 activation regulated by TIMP-3. / Yokoyama, Takeyoshi; Nakamura, Hiroyuki; Otani, Yoshihide; Kubota, Tetsuro; Fujimoto, Noboru; Seiki, Motoharu; Kitajima, Masaki; Okada, Yasunori.

In: Clinical and Experimental Metastasis, Vol. 21, No. 3, 2004, p. 223-233.

Research output: Contribution to journalArticle

Yokoyama, Takeyoshi ; Nakamura, Hiroyuki ; Otani, Yoshihide ; Kubota, Tetsuro ; Fujimoto, Noboru ; Seiki, Motoharu ; Kitajima, Masaki ; Okada, Yasunori. / Differences between scirrhous and non-scirrhous human gastric carcinomas from the aspect of proMMP-2 activation regulated by TIMP-3. In: Clinical and Experimental Metastasis. 2004 ; Vol. 21, No. 3. pp. 223-233.
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AU - Otani, Yoshihide

AU - Kubota, Tetsuro

AU - Fujimoto, Noboru

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AU - Kitajima, Masaki

AU - Okada, Yasunori

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