Chahwala S B, Cantley L C
J Biol Chem. 1984 Nov 25;259(22):13717-22.
Extracellular ATP (1 mM) inhibited the growth of Friend virus-infected murine erythroleukemia cells (MEL cells) but had no effect on dimethyl sulfoxide-induced differentiation. ATP (1 mM) also caused changes in the permeability of MEL cells to ions. There was an increased influx of 45Ca2+ from a basal level of 5 pmol/min to 18 pmol/min/10(6) cells to achieve a 2-fold increase in steady-state Ca2+ as measured at isotopic equilibration. Ca2+ influx was blocked by diisothiocyanostilbene disulfonate (DIDS), an inhibitor of anion transport. ATP also stimulated Cl- uptake, and this flux was inhibited by DIDS. The ratio of ATP stimulated Cl- to Ca2+ uptake was 1.6:1. K+ and Na+ influx were also stimulated by ATP, but phosphate uptake was inhibited; the Na+ influx dissipated the Na+ gradient and thus inhibited nutrient uptake. ATP-stimulated K+ influx was ouabain inhibitable; however, the total cellular K+ decreased due to an ATP-stimulated ouabain-resistant K+ efflux. Na+ influx and Ca2+ influx occurred by separate independent routes, since Na+ influx was not inhibited by DIDS. The effects observed were specific for ATP *K1/2 MgATP = 0.7 mM) since AMP, GTP, adenosine, and the slowly hydrolyzable ATP analogue adenyl-5'-yl imidodiphosphate were without effect. The major ionic changes in the cell were a decrease in K+ and increase in Na+; cytoplasmic pH and free Ca2+ did not change appreciably. These ATP-induced changes in ion flux are considered to be responsible for growth inhibition.
细胞外ATP(1 mM)可抑制感染了Friend病毒的小鼠红白血病细胞(MEL细胞)的生长,但对二甲基亚砜诱导的分化没有影响。ATP(1 mM)还会引起MEL细胞对离子的通透性变化。45Ca2+的流入量从基础水平的5 pmol/分钟增加到18 pmol/分钟/10(6)个细胞,在同位素平衡时测量的稳态Ca2+增加了2倍。Ca2+的流入被阴离子转运抑制剂二异硫氰酸芪二磺酸(DIDS)阻断。ATP还刺激了Cl-的摄取,并且这种通量被DIDS抑制。ATP刺激的Cl-与Ca2+摄取的比率为1.6:1。ATP也刺激了K+和Na+流入,但磷酸盐摄取受到抑制;Na+流入消耗了Na+梯度,从而抑制了营养物质的摄取。ATP刺激的K+流入可被哇巴因抑制;然而,由于ATP刺激的哇巴因抗性K+外流,细胞内总K+减少。Na+流入和Ca2+流入通过不同的独立途径发生,因为Na+流入不受DIDS抑制。观察到的这些效应是ATP特有的(K1/2 MgATP = 0.7 mM),因为AMP、GTP、腺苷和水解缓慢的ATP类似物腺苷-5'-基亚氨基二磷酸没有作用。细胞内主要的离子变化是K+减少和Na+增加;细胞质pH和游离Ca2+没有明显变化。这些ATP诱导的离子通量变化被认为是生长抑制的原因。