Olson J E, Pledger W J
Department of cell Biology, Vanderbilt University, Nashville, Tennessee 37232.
J Biol Chem. 1990 Feb 5;265(4):1847-51.
The addition of platelet-derived growth factor (PDGF) to many types of cells causes a rapid decrease in high affinity binding of 125I-epidermal growth factor (EGF), a process which has been termed transmodulation. Treatment with the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA) also results in the transmodulation of the EGF receptor in many cell types. PDGF can transmodulate EGF binding through a mechanism that is not dependent on protein kinase C activity. A recent report (Wattenberg, E. V., McNeil, P. L., Fujiki, H., and Rosner, M. R. (1989) J. Biol. Chem. 264, 213-219) described the requirement for a sodium ion influx in the down-modulation of the EGF receptor stimulated by a non-TPA-type tumor promoter, palytoxin, in Swiss 3T3 cells. We tested for a similar sodium requirement in Balb/c/3T3 and Swiss 3T3 cells stimulated by PDGF or TPA in Balb cells treated with TPA for prolonged periods to down-regulate protein kinase C activity. Our results clearly show that the PDGF- and TPA-stimulated transmodulation of the EGF receptor does not require external sodium nor is the process affected by amiloride. In each of these experiments, the loss of 125I-EGF binding occurred to a similar extent and at a similar rate in the presence or absence of sodium. Intracellular pH also did not appear to have a role in the response. The sodium ionophore, monensin, was previously shown to bring about the down-modulation of 125I-EGF binding in Swiss cells. However, our results indicate that monensin-induced transmodulation of the EGF receptor occurs with or without external sodium, suggesting that the loss of binding is not the result of a sodium ion influx. These findings demonstrate that an increase in intracellular sodium does not cause nor is it required for PDGF- or TPA-stimulated EGF receptor transmodulation.
在多种细胞类型中添加血小板衍生生长因子(PDGF)会导致125I-表皮生长因子(EGF)的高亲和力结合迅速减少,这一过程被称为转调。用肿瘤启动子12-O-十四烷酰佛波醇-13-乙酸酯(TPA)处理也会在许多细胞类型中导致EGF受体的转调。PDGF可通过一种不依赖蛋白激酶C活性的机制转调EGF结合。最近的一份报告(Wattenberg, E. V., McNeil, P. L., Fujiki, H., and Rosner, M. R. (1989) J. Biol. Chem. 264, 213 - 219)描述了在瑞士3T3细胞中,非TPA型肿瘤启动子刺尾鱼毒素刺激EGF受体下调时对钠离子内流的需求。我们在经TPA长期处理以下调蛋白激酶C活性的Balb/c/3T3细胞和瑞士3T3细胞中,检测了PDGF或TPA刺激下是否有类似的钠需求。我们的结果清楚地表明,PDGF和TPA刺激的EGF受体转调不需要细胞外钠,该过程也不受氨氯吡咪影响。在这些实验的每一个中,无论有无钠,125I-EGF结合的丧失都以相似的程度和相似的速率发生。细胞内pH似乎在该反应中也不起作用。钠离子载体莫能菌素先前已被证明可导致瑞士细胞中125I-EGF结合的下调。然而,我们的结果表明,无论有无细胞外钠,莫能菌素诱导的EGF受体转调都会发生,这表明结合的丧失不是钠离子内流的结果。这些发现表明,细胞内钠的增加既不会导致PDGF或TPA刺激的EGF受体转调,也不是其必需条件。