Taylor M V, Hesketh T R, Metcalfe J C
Department of Biochemistry, Cambridge, U.K.
Biochem J. 1988 Feb 1;249(3):847-55. doi: 10.1042/bj2490847.
Comparisons were made between transformed S49 T-lymphoma cells and normal murine thymocytes in their polyphosphoinositides, inositol polyphosphates and cytosolic free calcium concentrations ([Ca2+]i), and the effects of the T-cell mitogen concanavalin A (Con A) on these properties. 1. The ratios of the polyphosphoinositides to phosphatidylinositol in both exponential-phase S49 cells and mitogen-stimulated thymocytes (G1 phase) were greater than in quiescent (G0-phase) thymocytes. 2. In response to Con A, the amount of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) in S49 cells decreased slightly (17% in 30 min), and this was sufficient to account for the small amounts of inositol phosphates that accumulated. In contrast, it has been shown previously that Con A stimulates a rapid resynthesis of PtdInsP2 in thymocytes and the amounts of inositol phosphates released rapidly exceed the steady-state amount of the PtdInsP2 precursor [Taylor, Metcalfe, Hesketh, Smith & Moore (1984) Nature (London) 312, 462-465]. 3. The [Ca2+]i did not differ significantly in S49 cells and thymocytes before the addition of Con A, and the increases in [Ca2+]i in response to Con A were similar in both types of cell. 4. The [Ca2+]i increase in response to Con A was inhibited by similar concentrations of intracellular cyclic AMP (2-10 microM) in S49 cells and thymocytes, suggesting that similar regulatory mechanisms act on this response in both types of cell. The data demonstrate that the basal [Ca2+]i and phosphoinositide metabolism is similar in both the normal cells and their transformed counterparts. In addition, they suggest that the activated Con A receptors generate very similar signals in the two cell types, and that any perturbations of primary signal transduction to the secondary phosphoinositide and [Ca2+]i responses in the S49 phenotype are quantitative rather than qualitative.
对转化的S49 T淋巴瘤细胞和正常小鼠胸腺细胞的多磷酸肌醇、肌醇多磷酸及胞质游离钙浓度([Ca2+]i)进行了比较,并研究了T细胞有丝分裂原伴刀豆球蛋白A(Con A)对这些特性的影响。1. 处于指数生长期的S49细胞和有丝分裂原刺激的胸腺细胞(G1期)中多磷酸肌醇与磷脂酰肌醇的比率均高于静止(G0期)胸腺细胞。2. 对Con A的反应中,S49细胞中磷脂酰肌醇4,5-二磷酸(PtdInsP2)的量略有下降(30分钟内下降17%),这足以解释积累的少量肌醇磷酸。相比之下,先前已表明Con A刺激胸腺细胞中PtdInsP2的快速重新合成,释放的肌醇磷酸量迅速超过PtdInsP2前体的稳态量[泰勒、梅特卡夫、赫斯基思、史密斯和摩尔(1984年)《自然》(伦敦)312, 462 - 465]。3. 添加Con A之前,S49细胞和胸腺细胞中的[Ca2+]i无显著差异,两种细胞类型对Con A反应时[Ca2+]i的增加相似。4. S49细胞和胸腺细胞中,相似浓度的细胞内环磷酸腺苷(2 - 10 microM)抑制了对Con A反应时[Ca2+]i的增加,表明相似的调节机制作用于两种细胞的这种反应。数据表明正常细胞及其转化对应物的基础[Ca2+]i和磷酸肌醇代谢相似。此外,它们表明活化的Con A受体在两种细胞类型中产生非常相似的信号,并且S49表型中从初级信号转导至次级磷酸肌醇和[Ca2+]i反应的任何扰动都是定量的而非定性的。