Bilal Fatima, Pérès Michaël, Andrieu-Abadie Nathalie, Levade Thierry, Badran Bassam, Daher Ahmad, Ségui Bruno
INSERM UMR 1037, CRCT, 31037, Toulouse, France.
Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France.
Methods Mol Biol. 2017;1557:207-212. doi: 10.1007/978-1-4939-6780-3_19.
Sphingomyelin synthases 1 and 2 convert the anti-oncometabolite ceramide to sphingomyelin, the most abundant sphingolipid in plasma membrane. CD95L-induced ceramide increase is associated with the caspase-dependent inhibition of sphingomyelin synthesis, which enhances the mitochondrial route to apoptosis. Knocking down sphingomyelin synthase 1 or inhibiting sphingomyelin synthesis facilitates ceramide accumulation, cytochrome c release from mitochondria, and caspase-9 activation in cancer cell upon CD95L treatment. Here, we describe a method to monitor in situ sphingomyelin synthase activity changes triggered by CD95L.
鞘磷脂合成酶1和2将抗癌代谢物神经酰胺转化为鞘磷脂，鞘磷脂是质膜中含量最丰富的鞘脂。CD95L诱导的神经酰胺增加与鞘磷脂合成的半胱天冬酶依赖性抑制有关，这增强了线粒体凋亡途径。敲低鞘磷脂合成酶1或抑制鞘磷脂合成可促进神经酰胺积累、线粒体细胞色素c释放以及CD95L处理后癌细胞中半胱天冬酶-9的激活。在此，我们描述了一种监测CD95L触发的原位鞘磷脂合成酶活性变化的方法。
