Mills B, Tupper J T
J Cell Physiol. 1976 Sep;89(1):123-32. doi: 10.1002/jcp.1040890112.
K transport has been investigated during progression of cultured Ehrlich ascites tumor cells through the cell cycle. Using a double thymidine block technique, Ehrlich cells carried in continuous culture have been synchronized, as verified by simultaneous monitoring of cell number, cell volume, 3H-thymidine incorporation and mitotic index. Unidirectional influx, efflux and cell content of K have been monitored throughout the cell cycle. The nature of the pump mediated, ouabain-sensitive K flux and the furosemide-sensitive component of K flux, presumably representing K-K exchange, have also been evaluated. In early S period the ouabain sensitive component, representing the Na-K pump, comprises 52% of the total unidirectional K influx and subsequently declines during G2 period to a minimum of 40% in mid G2. During M and early S the activity again rises. As the ouabain sensitive component becomes maximal in late S period, the furosemide sensitive component diminishes from approximately 30% of the total influx to approximately 10%. The same pattern is observed in the G2 period. As the pump component diminishes, the furosemide sensitive component increases. Furosemide sensitive K efflux has also been monitored and the pattern is equivalent to that observed in the invlux studies. No change in net K flux is observed in the presence of furosemide. This indicates that the furosemide sensitive component represents an exchange component for K. These results are consistent with the conclusion that the alterations in exchange and pump fluxes are physiological events associated with progression of the cell cycle.
在培养的艾氏腹水瘤细胞通过细胞周期的过程中,对钾转运进行了研究。采用双胸腺嘧啶核苷阻断技术,对连续培养的艾氏细胞进行同步化处理,通过同时监测细胞数量、细胞体积、³H-胸腺嘧啶核苷掺入量和有丝分裂指数进行验证。在整个细胞周期中监测了钾的单向流入、流出和细胞内含量。还评估了泵介导的、哇巴因敏感的钾通量的性质以及钾通量中呋塞米敏感成分(可能代表钾-钾交换)。在S期早期,代表钠-钾泵的哇巴因敏感成分占单向钾流入总量的52%,随后在G2期下降,在G2中期降至最低的40%。在M期和S期早期,该活性再次升高。当哇巴因敏感成分在S期后期达到最大值时,呋塞米敏感成分从总流入量的约30%降至约10%。在G2期观察到相同的模式。随着泵成分减少,呋塞米敏感成分增加。还监测了呋塞米敏感的钾流出,其模式与流入研究中观察到的模式相同。在存在呋塞米的情况下,净钾通量未观察到变化。这表明呋塞米敏感成分代表钾的交换成分。这些结果与以下结论一致,即交换通量和泵通量的变化是与细胞周期进程相关的生理事件。