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転移性を獲得した細胞の軟寒天中の行動と細胞接着及び細胞骨格の関係

软琼脂中获得转移特性的细胞行为、细胞粘附和细胞骨架之间的关系

基本信息

批准号：
06670218
负责人：
NOMURA Takahiro
金额：
$ 1.34万
依托单位：
Kanazawa 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：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670218/
关键词：
ras myc SFME r mHM-SFME-1 metastasis Balb c actin-microfilaments soft agar colony cell adhesion SFME細胞 mycSFME細胞 mHM-SFME-1細胞 cマウス腫瘍

项目摘要

Distribution of actin-microfilaments in two cell lines, one is ras/mycSFME and the other is r/mHM-SFME-1, was studied in a colony forming conditions in agar gel. The later, which derived from former, shows highly metastatic potentials in host animals whereas the former shows lower activity. Both cells were transformed by human c-Ha-ras and mouse c-myc genes and produced colonies in soft agar. The r/mHM-SFME-1 colonies were consisted of loosely packed cells with dispersed satellite cells around the colony whereas ras/mycSFME ones were made of fairly compacted cells. Rhodamine-phalloidin staining showed that microfilaments of the two cells were present mainly at cell periphery. The distribution of microfilaments in this condition was more abundant in ras/mycSFME cells than in r/mHM-SFME-1 cells. The cell-cell adhesion developed well in former than in the later one.Actin-microfilaments were detected mainly at the cell-cell contacted area in ras/mycSFME cells, whereas they were forming thin network under the surface of r/mHM-SFME-1 cells. When these cells were cultured on fibronectin coated dishes, however, they expressed a fibrobrastic appearance and spread well on the dishes. Then, the phenotypic difference in metastatic potentials of the two cells could be demonstrated in the three-dimensional cultures in soft agar gel.

在琼脂凝胶集落形成条件下，研究了肌动蛋白微丝在ras/mycSFME和r/mHM-SFME-1细胞系中的分布。后者源于前者，在宿主动物中具有高度转移潜力，而前者的活性较低。两种细胞分别被人c-Ha-ras和小鼠c-myc基因转化，并在软琼脂中产生菌落。r/mHM-SFME-1菌落由松散排列的细胞组成，周围有分散的卫星细胞，而ras/mycSFME菌落则由相当紧凑的细胞组成。罗丹明-phalloidin染色显示两种细胞的微丝主要存在于细胞周围。在这种情况下，ras/mycSFME细胞的微丝分布比r/mHM-SFME-1细胞的微丝分布更丰富。前者的细胞粘附较后者好。在ras/mycSFME细胞中，肌动蛋白微丝主要在细胞-细胞接触区检测到，而在r/mHM-SFME-1细胞中，肌动蛋白微丝在细胞表面下形成薄的网状结构。然而，当这些细胞在涂有纤维连接蛋白的培养皿中培养时，它们表现出成纤维细胞的外观，并在培养皿中很好地扩散。然后，在软琼脂凝胶三维培养中，可以证明两种细胞转移潜能的表型差异。