Mechanisms in metastasis of hepatocellular carcinoma and analysis of stimulative molecules

肝细胞癌转移机制及刺激分子分析

基本信息

批准号：
02670307
负责人：
HIGASHI Toshihiro
金额：
$ 1.22万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670307/
关键词：
Hepatocellular carcinoma Melanoma Metastasis Stimulative molecule for metastasis Type IV collagen degrading enzyme Invasion chamber ICG GST-alpha メラノ-マ IV型コラ-ゲン分解酵素 GSTーα

项目摘要

I)In vitro model of invasion and metastasis of cultured Hepatoma cells by using the invasion chamber.The potentiality of hepatoma cells(HuH-7 and PLC/PRF/5) was estimated as follows. We calculated cell numbers in the lower chamber which passed through the 13um pore size chemotaxis filter coated with 50ug of reconstituted matrix gel. The both hepatoma cells did not pass through the chemotaxis filter by themselves. On the contrary, mouse melanoma cells(B16 melanoma cell line) passed through the filter and we recognized that the cells entered into and destroyed the gel. Hepatoma cells could pass through the filter by adding culture medium of melanoma to medium of hepatoma in the upper chamber, indicating that melanoma cells secreted molecules for invasion and metastasis.II)Analysis of the molecules which stimulate invasion and metastasis of hepatoma cellsType IV collagen degrading enzyme activity was recognized in the culture medium of both melanoma and PLC/PRF/5. However, this enzyme was found as an inactive form which was activated by APMA. Sephacryl-S 200 gel filtration revealed that the molecular weight of the molecule was 37kD, suggesting that this molecule was different from autocrine motility factor, type IV collagen degrading enzyme and basic FGF.III)Functional diagnosis of hepatocellular carcinomaIn order to use clinical samples, we established functional diagnostic method for hepatoma. Aspiration biopsy after indocyanine green(ICG) was found as a useful device in which more than 95% of ICG negative tissues were malignant cells. We clarified that the mechanism of ICG loss was partly due to the decrease of cytoplasmic glutathion-S-transferase.

i）通过使用入侵室的培养肝癌细胞的体外侵袭和转移模型。估计肝癌细胞的潜力（HuH-7和PLC/PRF/5）的潜力如下。我们计算了下部腔室中的细胞数量，该腔室通过了涂有50倍重构的基质凝胶的13UM孔趋化滤光圈。两种肝癌细胞都没有通过趋化滤光度过滤。相反，小鼠黑色素瘤细胞（B16黑色素瘤细胞系）通过过滤器，我们认识到细胞进入并破坏了凝胶。肝癌细胞可以通过将黑色素瘤的培养基添加到上腔中的肝瘤培养基中，表明黑色素瘤细胞分泌的分子用于浸润和转移。但是，该酶是被APMA激活的非活性形式。 Sephacryl-S 200凝胶过滤表明该分子的分子量为37KD，这表明该分子与自分泌运动因子不同，IV型胶原蛋白降解酶和基本FGF.III）用于诊断临床诊断，以诊断为诊断术的肝癌疗法。发现吲哚烷绿色（ICG）后的抽吸活检是一种有用的装置，其中超过95％的ICG阴性组织是恶性细胞。我们阐明了ICG损失的机制部分是由于细胞质谷胱甘肽-S-转移酶的减少。