从注射了LM成纤维细胞的C3H小鼠培养的转移性和非转移性细胞的膜特性。

Department of Pathology and Laboratory Medicine, College of Medicine, University of Cincinnati, OH 45267-0529.

Biochim Biophys Acta. 1990 Mar;1022(3):365-72. doi: 10.1016/0005-2736(90)90287-x.

Little is known regarding the membrane properties of metastatic cells as compared to non-metastatic tumor cells. In order to remove variables such as site of growth and nutrition, C3H mice and LM fibroblasts were used as a model system to derive cell lines from local tumors and lung metastases. LM cells were injected subcutaneously into C3H mice and local skin tumors and secondary lung tumors were isolated, cultured in vitro and analyzed. The activities of lipid-sensitive membrane enzymes, membrane lipid composition, and membrane structure were correlated with metastatic ability. Plasma membranes and microsomes of the cultured metastatic cells had 3.8 +/- 0.5- and 5.4 +/- 0.6-fold elevated 5'-nucleotidase activity, respectively, as compared to plasma membranes and microsomes of cultured non-metastatic cells. The mitochondria of cultured metastatic cells had 3.5 +/- 0.5-fold decreased succinate-dependent cytochrome-c reductase activity as compared to mitochondria of the cultured non-metastatic cells. The lipids of plasma membranes from the metastatic cells had 30 +/- 2% and 46 +/- 7% lower phosphatidylinositol and sterol/phospholipid ratio, respectively, and 30 +/- 3% increased unsaturated/saturated fatty acid as compared to cultured non-metastatic cells. The lower sterol/phospholipid ratio correlated with a 30 +/- 1% lower level of cytosolic sterol carrier protein in the cultured metastatic cells as compared to cultured non-metastatic cells. Multifrequency phase and modulation fluorometry in conjunction with the fluorescence probe, 1,6-diphenyl-1,3,5-hexatriene, was used to determine the static and dynamic aspects of membrane fluidity. The plasma membranes and microsomes of cultured metastatic cells were more fluid than those of cultured non-metastatic cells as indicated by 24 +/- 3% and 7 +/- 1%, respectively, lower limiting anisotropy of 1,6-diphenyl-1,3,5-hexatriene in the membranes of the metastatic as compared to non-metastatic cells.

与非转移性肿瘤细胞相比，关于转移性细胞的膜特性我们所知甚少。为了消除诸如生长部位和营养等变量的影响，采用C3H小鼠和LM成纤维细胞作为模型系统，从局部肿瘤和肺转移瘤中获取细胞系。将LM细胞皮下注射到C3H小鼠体内，分离出局部皮肤肿瘤和继发性肺肿瘤，进行体外培养并分析。脂质敏感膜酶的活性、膜脂质组成和膜结构与转移能力相关。与培养的非转移性细胞的质膜和微粒体相比，培养的转移性细胞的质膜和微粒体的5'-核苷酸酶活性分别提高了3.8±0.5倍和5.4±0.6倍。与培养的非转移性细胞的线粒体相比，培养的转移性细胞的线粒体的琥珀酸依赖性细胞色素c还原酶活性降低了3.5±0.5倍。与培养的非转移性细胞相比，转移性细胞质膜的脂质中磷脂酰肌醇和甾醇/磷脂比率分别降低了30±2%和46±7%，不饱和/饱和脂肪酸增加了30±3%。与培养的非转移性细胞相比，培养的转移性细胞中较低的甾醇/磷脂比率与胞质甾醇载体蛋白水平降低30±1%相关。结合荧光探针1,6-二苯基-1,3,5-己三烯，使用多频相位和调制荧光测定法来确定膜流动性的静态和动态方面。与非转移性细胞相比，转移性细胞的质膜和微粒体的流动性更高，这表现为在转移性细胞膜中1,6-二苯基-1,3,5-己三烯的极限各向异性分别降低了24±3%和7±1%。