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在肝细胞癌的经门脉肝内转移及远位转移时,癌细胞首先必须破坏血管的基底膜。Ⅳ型胶原分解酶,可特异性地对构成基底膜成分的Ⅳ型胶原进行分解。本酶活性在痴变部位呈高值,对有明显远位转移倾向的病例,在癌部位的活性也颇高。作者等对肝癌患者的该酶在血清中的活性及检测可能性作了探讨。对象:50～70,岁男性24例,其中健康人、(对照组)7例,冈山大学第一内科确诊入院的肝硬化组(LC组)7例,肝硬化并肝细胞癌组(HCC+LC组)10例。方法:为使该酶激活,在60μl血清中加入1mM 的4—氨苯汞醋酸盐(APMA),于37℃置2小时,加入435μg 的胰蛋白酶放置5分钟作预培养,再用大豆胰蛋白酶抑制剂使胰蛋白酶久活,留作酶溶液。以取自人胎盘经精制标识的Ⅳ型胶原作为基质,加入含CaCl_2、NaCl、异丙氟磷.APMA 的50mM Tris-HCl 缓冲溶液(pH7.5)混合培养后,测定分解后的Ⅳ型胶原放射活性。并存各酶溶液中,加入EDTA(依地酸)同
In portal hepatic metastasis and distant metastasis of hepatocellular carcinoma, cancer cells must first destroy the basement membrane of the blood vessel. Type IV collagenase can specifically decompose type IV collagen that constitutes a basement membrane component. The activity of this enzyme is high at the site of dementia, and it is also highly active at the site of cancer in patients with a tendency to metastasize significantly. The authors discussed the activity of the enzyme in liver cancer patients and the possibility of detection. Subjects: 50 to 70 years old, 24 males, including healthy subjects (control group), 7 cases, admitted to the first internal medicine department of Okayama University, 7 cases of liver cirrhosis group (LC group), liver cirrhosis and hepatocellular carcinoma group (HCC +LC group) 10 cases. METHODS: To activate the enzyme, 1 mM 4-Aminophenylmercuric acetate (APMA) was added to 60 μl of serum, set at 37°C for 2 hours, and 435 μg of trypsin was added for 5 minutes for preculture, followed by soybean pancreas. Protease inhibitors allow the trypsin to live for a long time and remain as an enzyme solution. Using type IV collagen from the human placenta as a matrix, mixed with 50 mM Tris-HCl buffer solution (pH 7.5) containing CaCl 2 , NaCl, and isopropyl fluorophosphonate (APMA), and measuring the decomposed type IV collagen. Radioactivity. Coexist in each enzyme solution and add EDTA