Potentiation of cannabinoid-induced cytotoxicity in mantle cell lymphoma through modulation of ceramide metabolism.
Potentiation of cannabinoid-induced cytotoxicity in mantle cell lymphoma through modulation of ceramide metabolism.
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DOI：
10.1158/1541-7786.mcr-08-0361
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发表时间：
2009-07
期刊：
影响因子：
--
通讯作者：
Flygare J
中科院分区：
文献类型：
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作者：
Gustafsson K;Sander B;Bielawski J;Hannun YA;Flygare J
经大麻素治疗后，外套细胞淋巴瘤细胞中神经酰胺水平升高。在这里，我们使用稳定的内源性大麻素类似物R(+)-甲烷胺(R-MA)研究神经酰胺在MCL细胞系Rec-1中积累的途径。我们进一步干扰了神经酰胺转化为神经鞘脂类促进细胞生长的过程。用R-MA处理后，神经酰胺C16、C18、C24和C24：1的水平增加，神经酰胺合成酶(CerSs)3和6的转录诱导增加。与大麻素受体1(CB1)有高亲和力的SR141716A可减弱这种作用。用Win-55,212-2证实了CB1介导的CerS3和CerS6 mRNA的诱导。使用siRNA同时沉默CerS3和CerS6可以消除R-MA诱导的C16和C24的积累。抑制从头神经酰胺途径中的丝氨酸棕榈酰转移酶、神经酰胺合成酶和二氢神经酰胺脱饱和酶，可以逆转R-MA处理诱导的神经酰胺积累和细胞死亡。为了增强R-MA、鞘氨醇-1(SK-1)和葡萄糖神经酰胺合成酶(GCS)的细胞毒作用，神经酰胺转化为促进增殖的鞘氨醇-1-磷酸和葡萄糖神经酰胺的酶分别被抑制。使用抑制剂或siRNA抑制酶会加剧R-MA处理引起的存活率下降、诱导细胞死亡和神经酰胺积累。我们的发现表明，R-MA通过CB1介导的从头神经酰胺合成途径上调而诱导MCL细胞死亡。此外，抑制SK-1和GCS可增强R-MA诱导的神经酰胺蓄积和细胞死亡。这是第一个通过调节神经酰胺代谢来增强大麻素的细胞毒作用的研究。
Ceramide levels are elevated in Mantle Cell Lymphoma cells following treatment with cannabinoids. Here, we investigated the pathways of ceramide accumulation in the MCL cell line Rec-1 using the stable endocannabinoid analogue R(+)-methanandamide (R-MA). We further interfered with the conversion of ceramide into sphingolipids that promote cell growth. Treatment with R-MA led to increased levels of ceramide species C16, C18, C24 and C24:1 and transcriptional induction of ceramide synthases (CerSs) 3 and 6. The effects were attenuated using SR141716A, which has high affinity to cannabinoid receptor 1 (CB1). The CB1-mediated induction of CerS3 and CerS6 mRNA was confirmed using Win-55,212-2. Simultaneous silencing of CerS3 and CerS6 using siRNA abrogated the R-MA-induced accumulation of C16 and C24. Inhibition of either of the enzymes serine palmiotyl transferase, ceramide synthase, and dihydroceramide desaturase within the de novo ceramide pathway reversed ceramide accumulation and cell death induced by R-MA treatment. In order to enhance the cytotoxic effect R-MA, sphingosine kinase-1 (SK-1) and glucosylceramide synthase (GCS), enzymes that convert ceramide to the pro-proliferative sphingolipids sphingosine-1-phospate and glucosylceramide, respectively, were inhibited. Suppression of either enzyme using inhibitors or siRNA potentiated the decreased viability, induction of cell death and ceramide accumulation induced by R-MA treatment. Our findings suggest that R-MA induces cell death in MCL via CB1-mediated upregulation of the de novo ceramide synthesis pathway. Furthermore, inhibition of SK-1 and GCS potentiated ceramide accumulation and cell death induced by R-MA. This is the first study were the cytotoxic effect of a cannabinoid is enhanced by modulation of ceramide metabolism.