Dong Z, Venkatachalam M A, Wang J, Patel Y, Saikumar P, Semenza G L, Force T, Nishiyama J
Department of Pathology, University of Texas Health Science Center, San Antonio, Texas 78229, USA.
J Biol Chem. 2001 Jun 1;276(22):18702-9. doi: 10.1074/jbc.M011774200. Epub 2001 Mar 12.
Hypoxia is a key determinant of tissue pathology during tumor development and organ ischemia. However, little is known regarding hypoxic regulation of genes that are directly involved in cell death or death resistance. Here we report the striking induction by severe hypoxia of the anti-apoptotic protein IAP-2. Hypoxic cells with IAP-2 up-regulation became resistant to apoptosis. IAP-2 was induced by hypoxia per se rather than by the secondary effects of hypoxia, including ATP depletion and cell injury. The inductive response did not relate to alterations of cellular redox status or arrest of mitochondrial respiration. On the other hand, IAP-2 induction was attenuated by actinomycin D, suggesting a role for gene transcription. In vitro nuclear run-on assays demonstrated specific increases in IAP-2 transcriptional activity after hypoxia exposure. HIF-1, the primary transcription factor that is responsible for multiple gene activation under hypoxia, does not have a role in IAP-2 expression. HIF-1 and IAP-2 were induced by different degrees of hypoxia; severe hypoxia or anoxia was required for IAP-2 induction. Moreover, cobalt chloride and desferrioxamine activated HIF-1 but not IAP-2. Finally, IAP-2 was induced by severe hypoxia in mouse embryonic stem cells that were deficient of HIF-1. Thus, this study not only provides the first demonstration of hypoxic regulation of an anti-apoptotic gene but also suggests the participation of novel hypoxia-responsive transcription mechanisms.
缺氧是肿瘤发展和器官缺血过程中组织病理学的关键决定因素。然而,对于直接参与细胞死亡或抗死亡的基因的缺氧调节知之甚少。在此,我们报告了严重缺氧对抗凋亡蛋白IAP-2的显著诱导作用。IAP-2上调的缺氧细胞对凋亡产生抗性。IAP-2是由缺氧本身诱导的,而非缺氧的继发效应,包括ATP耗竭和细胞损伤。诱导反应与细胞氧化还原状态的改变或线粒体呼吸的停滞无关。另一方面,放线菌素D可减弱IAP-2的诱导作用,提示基因转录发挥了作用。体外核转录分析表明,缺氧暴露后IAP-2转录活性有特异性增加。HIF-1是负责缺氧条件下多个基因激活的主要转录因子,在IAP-2表达中不起作用。HIF-1和IAP-2受不同程度缺氧的诱导;IAP-2的诱导需要严重缺氧或无氧条件。此外,氯化钴和去铁胺激活HIF-1但不激活IAP-2。最后,在缺乏HIF-1的小鼠胚胎干细胞中,严重缺氧可诱导IAP-2。因此,本研究不仅首次证明了抗凋亡基因的缺氧调节,还提示了新型缺氧反应转录机制的参与。