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

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.