Establishment of Quantitative Method for Measurement of MMPs in Blood and Bile, and Its Clinical Application

我们建立了定量测定血液和胆汁中MMP-2和MMP-9的方法。我们将此方法应用于临床样品，并发现了以下新的结果。（1）胃癌患者血清MMP-9水平显著高于正常对照组。MMP-9水平在I、II、III、IV期胃癌患者中以IV期最高。我们研究了MMP-9水平与胃癌患者生存的相关性，发现MMP-9水平（>410）是预测胃癌患者生存的良好指标。(2)We首次建立了测定胆汁中MMP-2和MMP-9的方法。此外，我们还发现，激活的MMP-2和proMMP-9活性是预测结肠癌患者肝转移的良好标志物，使用抗MMP-2抗体对各种癌组织切片进行免疫组化分析，我们发现，MMP-2在80种癌症中的59%的病例中表达，包括结肠、子宫、肺、肝、脑、软组织和食管。我们首次发现，MMP-2的表达是转录控制的ODC在癌细胞中使用凝胶迁移和启动子分析。由于ODC是一种原癌基因，在多种肿瘤中均有表达，因此抑制ODC的表达是治疗肿瘤和转移的有效方法。

We have established quantitative method for the measurement of MMPs, especially MMP-2 and MMP-9 in blood and bile. We applied this method to clinical samples, and found the following new results.(1)MMP-9 levels in blood was significantly elevated in gastric cancer patients, compared to normal healthy subjects. In stage IV gastric cancer patients MMP-9 level was the highest among stages I, II, III and IV.We examined the correlation between MMP-9 level and the survival of gastric cancer patients, and found that the level of MMP-9(>410)was a good indicator for predicting the survival of gastric cancer patients.(2)We for the first time established a method for measuring MMP-2 and MMP-9 in bile. Further, we have found that activated MMP-2 and proMMP-9 activities were good markers for predicting liver metastasis in colon cancer patients.Immunohistochemical analysis using anti-MMP-2 antibody was performed in various cancer tissue sections and we found that MMP-2 expressed in 59% cases out of 80 various cancer including colon, uterus, lung, liver brain, soft tissue and esophagus. We for the first time found that MMP-2 expression was transcriptionally controlled by ODC in cancer cells using gel shift and promotor assays. As ODC is now considered to be a protooncogene and expresses in various cancer, the results suggest that suppressing ODC expression is a reasonabale approach to treat cancer and metastasis.