Deregibus Maria Chiara, Cantaluppi Vincenzo, Doublier Sophie, Brizzi Maria Felice, Deambrosis Ilaria, Albini Adriana, Camussi Giovanni
Cattedra di Nefrologia, Dipartimento di Medicina Interna Università di Torino, Centro Ricerca Medicina Sperimentale (CeRMS), Italy.
J Biol Chem. 2002 Jul 12;277(28):25195-202. doi: 10.1074/jbc.M200921200. Epub 2002 May 6.
In this study we found that Tat protected vincristine-treated Kaposi's sarcoma cells from apoptosis and from down-regulation of several anti-apoptotic genes such as AKT-1, AKT-2, BCL2, BCL-XL, and insulin-like growth factor I and induced the de novo expression of the interleukin-3 gene. Moreover, we found that Tat enhanced phosphorylation of AKT and BAD proteins. The inhibition of phosphatidylinositol 3-kinase with two unrelated pharmacological inhibitors, wortmannin and LY294002, abrogated both the anti-apoptotic effect and the phosphorylation of AKT induced by Tat. After treatment with Tat, the AKT enzymatic activity showed a biphasic increase: an early activation (15 min), independent from protein synthesis; and a delayed activation (24 h), which was significantly decreased upon blockage of protein synthesis. Experiments with a function blocking anti-vascular endothelial cell growth factor receptor-2 antibody suggested that both the early and delayed AKT activation and the protection from apoptosis were triggered by the interaction of Tat with vascular endothelial cell growth factor receptor-2. Moreover, experiments with function-blocking antibodies directed against insulin-like growth factor I/insulin-like growth factor I receptor or interleukin-3 indicated their involvement in the delayed activation of AKT and their contribution to the anti-apoptotic effect of Tat on vincristine-treated Kaposi's sarcoma cells.
在本研究中,我们发现Tat可保护长春新碱处理的卡波西肉瘤细胞免于凋亡,并防止几种抗凋亡基因如AKT-1、AKT-2、BCL2、BCL-XL和胰岛素样生长因子I的下调,并诱导白细胞介素-3基因的从头表达。此外,我们发现Tat增强了AKT和BAD蛋白的磷酸化。用两种不相关的药理抑制剂渥曼青霉素和LY294002抑制磷脂酰肌醇3激酶,消除了Tat诱导的抗凋亡作用和AKT的磷酸化。用Tat处理后,AKT酶活性呈双相增加:早期激活(15分钟),与蛋白质合成无关;延迟激活(24小时),在蛋白质合成受阻时显著降低。用功能阻断抗血管内皮细胞生长因子受体-2抗体进行的实验表明,Tat与血管内皮细胞生长因子受体-2的相互作用触发了早期和延迟的AKT激活以及对凋亡的保护。此外,用针对胰岛素样生长因子I/胰岛素样生长因子I受体或白细胞介素-3的功能阻断抗体进行的实验表明,它们参与了AKT的延迟激活,并对Tat对长春新碱处理的卡波西肉瘤细胞的抗凋亡作用有贡献。