Huggins J W, Chesnut R W, Durham N N, Carraway K
Biochim Biophys Acta. 1976 Apr 5;426(4):630-7. doi: 10.1016/0005-2736(76)90127-9.
Sarcoma-180 tumor cells in culture or grown as an ascites form in the CD-1 mouse have been subjected to mild trypsinization procedures in order to study morphological and molecular changes resulting from proteolysis. The cells attached to a substratum become rounded within 20 min. and most undergo cell division, but they do not detach from the substratum. Removal of trypsin permits the cells to go back to their original spindle shape over an 8-20 h period. Surface membranes were isolated from trypsinized ascites and cultured cells and subjected to dodecyl sulfate-acrylamide gel electrophoresis. Both cell types showed the same two kinds of changes in electrophoretic patterns. First, there was a loss of glycoproteins from both cell types even though they show different complements of cell surface glycoproteins. Second, there is a loss of high molecular weight polypeptides, which have previously been suggested to play a role in membrane stabilization and cell shape. These results further implicate these polypeptides in the control of cell morphology and offer circumstanital evidence for transmembrane interactions of surface glycoproteins with the high molecular weight polypeptides as a factor in controlling cell morphology.
为了研究蛋白水解作用所导致的形态学和分子变化,已对培养中的肉瘤180肿瘤细胞或在CD-1小鼠体内以腹水形式生长的肉瘤180肿瘤细胞进行了温和的胰蛋白酶消化处理。附着在基质上的细胞在20分钟内变圆,大多数细胞会进行细胞分裂,但它们不会从基质上脱离。去除胰蛋白酶后,细胞会在8至20小时内恢复到原来的纺锤形。从经胰蛋白酶处理的腹水细胞和培养细胞中分离出表面膜,并进行十二烷基硫酸钠-丙烯酰胺凝胶电泳。两种细胞类型在电泳图谱上都显示出相同的两种变化。首先,尽管两种细胞类型的细胞表面糖蛋白组成不同,但它们都出现了糖蛋白的丢失。其次,则是高分子量多肽的丢失,此前有研究表明这些多肽在膜稳定和细胞形态方面发挥作用。这些结果进一步表明这些多肽参与细胞形态的控制,并为表面糖蛋白与高分子量多肽的跨膜相互作用作为控制细胞形态的一个因素提供了间接证据。