TY - JOUR
T1 - Differences between scirrhous and non-scirrhous human gastric carcinomas from the aspect of proMMP-2 activation regulated by TIMP-3
AU - Yokoyama, Takeyoshi
AU - Nakamura, Hiroyuki
AU - Otani, Yoshihide
AU - Kubota, Tetsuro
AU - Fujimoto, Noboru
AU - Seiki, Motoharu
AU - Kitajima, Masaki
AU - Okada, Yasunori
N1 - Funding Information:
We are grateful to Dr Kazushi Iwata, Daiichi Fine Chemical Co., Ltd, Takaoka, Japan, for providing us with monoclonal antibody to TIMP-3. We also thank Miss Michiko Uchiyama for her technical assistance. This work was supported by a grant-in-aid from the Ministry of Education, Science and Culture of Japan to Y.O.
PY - 2004
Y1 - 2004
N2 - 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.
AB - 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.
KW - MMP
KW - MT1-MMP
KW - ProMMP-2 activation
KW - TIMP-3
KW - scirrhous gastric carcinoma
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UR - http://www.scopus.com/inward/citedby.url?scp=4344689352&partnerID=8YFLogxK
U2 - 10.1023/B:CLIN.0000037704.72028.72
DO - 10.1023/B:CLIN.0000037704.72028.72
M3 - Article
C2 - 15387372
AN - SCOPUS:4344689352
VL - 21
SP - 223
EP - 233
JO - Clinical and Experimental Metastasis
JF - Clinical and Experimental Metastasis
SN - 0262-0898
IS - 3
ER -
