Stefanis L, Troy C M, Qi H, Greene L A
Department of Pathology, Taub Center for Alzheimer's Disease Research and Center for Neurobiology and Behavior, Columbia University College of Physicians and Surgeons, New York, New York 10032, U.S.A.
J Neurochem. 1997 Oct;69(4):1425-37. doi: 10.1046/j.1471-4159.1997.69041425.x.
Rat pheochromocytoma (PC12) cells and sympathetic neurons undergo apoptotic cell death upon withdrawal of trophic support. We have shown previously that selective cysteine aspartase (caspase) inhibitors protect PC12 cells and sympathetic neurons from such death, and that the caspase Nedd-2 is required for this type of death to occur. We now show that 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) and N(alpha)-p-tosyl-L-lysine chloromethyl ketone (TLCK), agents that inhibit another class of proteases, the trypsin-like serine proteases, also suppress cell death in this paradigm. The site of action of these agents is upstream of the caspases, because the CPP32-like and Nedd-2-cleaving activities that are induced upon withdrawal of trophic support in PC12 cells are inhibited when AEBSF and TLCK are applied to the cells. Both agents inhibit thymidine incorporation in PC12 cells at concentrations similar to those that promote survival, raising the possibility that they may promote survival in neuronal cells through inhibition of aberrant activation of cell cycle components.
去除营养支持后，大鼠嗜铬细胞瘤（PC12）细胞和交感神经元会发生凋亡性细胞死亡。我们之前已经表明，选择性半胱天冬酶（caspase）抑制剂可保护PC12细胞和交感神经元免于此类死亡，并且caspase Nedd-2是这种类型死亡发生所必需的。我们现在表明，4-（2-氨基乙基）苯磺酰氟盐酸盐（AEBSF）和N-α-对甲苯磺酰-L-赖氨酸氯甲基酮（TLCK），这两种抑制另一类蛋白酶（胰蛋白酶样丝氨酸蛋白酶）的试剂，在这种模式下也能抑制细胞死亡。这些试剂的作用位点在半胱天冬酶的上游，因为当将AEBSF和TLCK应用于PC12细胞时，去除营养支持后诱导的CPP32样和Nedd-2切割活性会受到抑制。两种试剂在促进存活的浓度下均抑制PC12细胞中的胸苷掺入，这增加了它们可能通过抑制细胞周期成分的异常激活来促进神经元细胞存活的可能性。
