Denecker G, Vercammen D, Steemans M, Vanden Berghe T, Brouckaert G, Van Loo G, Zhivotovsky B, Fiers W, Grooten J, Declercq W, Vandenabeele P
Molecular Signaling and Cell Death Unit, Department of Molecular Biology, Flanders Interuniversity Institute for Biotechnology and Ghent University, 9000 Gent, Belgium.
Cell Death Differ. 2001 Aug;8(8):829-40. doi: 10.1038/sj.cdd.4400883.
In L929sAhFas cells, tumor necrosis factor (TNF) leads to necrotic cell death, whereas agonistic anti-Fas antibodies elicit apoptotic cell death. Apoptosis, but not necrosis, is correlated with a rapid externalization of phosphatidylserine and the appearance of a hypoploid population. During necrosis no cytosolic and organelle-associated active caspase-3 and -7 fragments are detectable. The necrotic process does not involve proteolytic generation of truncated Bid; moreover, no mitochondrial release of cytochrome c is observed. Bcl-2 overexpression slows down the onset of necrotic cell death. In the case of apoptosis, active caspases are released to the culture supernatant, coinciding with the release of lactate dehydrogenase. Following necrosis, mainly unprocessed forms of caspases are released. Both TNF-induced necrosis and necrosis induced by anti-Fas in the presence of the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone are prevented by the serine protease inhibitor N-tosyl-L-phenylalanine chloromethylketone and the oxygen radical scavenger butylated hydroxyanisole, while Fas-induced apoptosis is not affected.
在L929sAhFas细胞中，肿瘤坏死因子（TNF）会导致坏死性细胞死亡，而促凋亡抗Fas抗体则引发凋亡性细胞死亡。凋亡而非坏死与磷脂酰丝氨酸的快速外化以及亚二倍体群体的出现相关。在坏死过程中，未检测到胞质和细胞器相关的活性半胱天冬酶-3和-7片段。坏死过程不涉及截短型Bid的蛋白水解生成；此外，未观察到细胞色素c从线粒体释放。Bcl-2过表达会减缓坏死性细胞死亡的发生。在凋亡情况下，活性半胱天冬酶会释放到培养上清液中，这与乳酸脱氢酶的释放同时发生。坏死发生后，主要是未加工形式的半胱天冬酶被释放。丝氨酸蛋白酶抑制剂N-对甲苯磺酰-L-苯丙氨酸氯甲基酮和氧自由基清除剂叔丁基对羟基茴香醚可阻止TNF诱导的坏死以及在存在半胱天冬酶抑制剂苄氧羰基-Val-Ala-Asp(OMe)-氟甲基酮的情况下抗Fas诱导的坏死，而Fas诱导的凋亡则不受影响。
