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De Novo-Synthesized Ceramide is Involved in Cannabinoid-Induced Apoptosis

July 10, 2015 -

Cannabinoids, such as Tetrahydrocannabinol (THC), alter our vision, our cognition and memory. They can be a potential treatment for preventing the growth of cancer cells in our bodies. Ceramide is a lipid that regulates cellular processes including proliferation and apoptosis. It is shown through the de novo synthesis pathway that exposure of glioma cells to cannabinoids initiates the generation of two peaks of ceramide.  The first peak may involve neutral sphingomyelinase stimulation via the adaptor protein FAN during the first few minutes of the cannabinoid treatment. The second peak starts on day 3 of the treatment, when apoptosis of the glioma cells occurs.

Two subclones of C6 glioma cells, C6.9, sensitive to THC-induced apoptosis, and C6.4, resistant to THC-induced apoptosis, were used in an experiment.  The cells were first incubated with the same amount of DSMO in different conditions and then lipids were extracted, saponified, and incubated with E-coli, eventually resulting in some ceramide 1-phosphate.  ERK and PKB activity levels were determined by Western-blot analysis. Reactions of the cells were started with different chemicals and stopped 30 minutes later with 0.5 M NH4OH. Cells were then washed with ice-cold PBS and all the RNA was extracted.

Two major pathways: phingomyelin hydrolysis and ceramide synthesis de novo, may contribute to intracellular ceramide accumulation. Results reveal that none of desipramine, scyphostatin, or PMA, prevented THC-induced apoptosis of glioma cells. However, a barrier of ceramide synthesis de novo with L-cycloserine or fumonisin B1 not only prevented THC-induced death of C6.9 glioma cells, but also THC-induced long-term ceramide accumulation in these cells.

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