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Investigation of the metastatic mechanism by a new in vitro invasion asseay using monolayrs of vascular endothelial cells.

使用单层血管内皮细胞进行新的体外侵袭试验研究转移机制。

基本信息

批准号：
07671441
负责人：
ITOH Hideaki
金额：
$ 1.47万
依托单位：
University of Occupational and Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671441/
关键词：
invasion assay endothelial cell metastasis plasminogen activator matrix metalloproteinase c-met Siaryl Lewis^a E-cadherin

项目摘要

Metastasis is a complex process. The interaction between endothelial cells and cancer cells is very important in this process. We established an in vitro invasion model using monolayrs of endothelial cells for investigation of this interaction. In human renal cancer cell lines, highly metastatic cell lines were more invasive than their low metastatic counterpart in our in vitro model. Our in vitro invasion assay using calf pulumonary arterial endothelial cells would be useful to evaluate protease activities and colony formation during invasion. Further examination of other sets of low and highly metastatic carcinoma cell lines is required to evaluate this in vitro invasion assay system. Now, we are producing several cell lines which will be useful for our invasion assay. In human renal cancer cell lines, we indicated the increased mRNA expressions of t-PA,u-PA and c-met by morthern blot analysis. In human colon carcinoma cell lines, we indicated the increased mRNA expressions of u-PA,MMP-2 and c-met by northern blot analysis, and we also indicated the increased expression of Siaryl Lewis^a and decreased expression of E-cadherin at cell surface by flow-cytometric analysis. These results were reported at some conferences and papors.

转移是一个复杂的过程。在这个过程中，内皮细胞和癌细胞之间的相互作用非常重要。我们建立了一个体外侵袭模型，使用单层内皮细胞的调查这种相互作用。在我们的体外模型中，在人肾癌细胞系中，高转移细胞系比低转移细胞系更具侵袭性。我们在体外侵袭小牛肺动脉内皮细胞检测将是有用的，以评估蛋白酶活性和集落形成过程中的入侵。需要进一步检查其他组的低和高转移癌细胞系，以评估这种体外侵袭测定系统。现在，我们正在生产几种细胞系，这将有助于我们的入侵检测。在人肾癌细胞系中，我们通过Morphimblot分析发现t-PA、u-PA和c-met的mRNA表达增加。北方印迹分析显示，在人结肠癌细胞系中，u-PA、MMP-2和c-met的mRNA表达增加，流式细胞术分析显示细胞表面Siaryl刘易斯^a的表达增加和E-cadherin的表达减少。这些结果在一些会议和论文中作了报道。