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Method to Measure Sphingomyelin Synthase Activity Changes in Response to CD95L.

Fatima Bilal1,2,3, Michaël Pérès1,2,4, Nathalie Andrieu-Abadie1,2

  • 1INSERM UMR 1037, CRCT, 31037, Toulouse, France.

Methods in Molecular Biology (Clifton, N.J.)
|January 13, 2017
PubMed
Summary

Sphingomyelin synthases convert ceramide to sphingomyelin. Inhibiting this process, especially sphingomyelin synthase 1, enhances cancer cell apoptosis by increasing ceramide levels.

Keywords:
ApoptosisCeramideIn situ sphingomyelin synthase activitySphingomyelin

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Cancer Research

Background:

  • Sphingomyelin synthases (SMS1 and SMS2) convert ceramide to sphingomyelin, a key plasma membrane sphingolipid.
  • CD95L-induced ceramide increase can inhibit SMS activity, promoting apoptosis via the mitochondrial pathway.
  • Cancer cells treated with CD95L show enhanced apoptosis when SMS1 is reduced or SMS activity is inhibited.

Purpose of the Study:

  • To develop a method for monitoring sphingomyelin synthase activity in situ.
  • To investigate the impact of CD95L treatment on sphingomyelin synthase activity.

Main Methods:

  • Monitoring sphingomyelin synthase activity in situ.
  • Utilizing CD95L treatment in cancer cell models.

Main Results:

  • CD95L treatment triggers changes in sphingomyelin synthase activity.
  • Knockdown of SMS1 or inhibition of sphingomyelin synthesis leads to ceramide accumulation.
  • This accumulation results in cytochrome c release and caspase-9 activation, promoting apoptosis.

Conclusions:

  • Sphingomyelin synthase activity is modulated by CD95L.
  • Targeting sphingomyelin synthesis is a potential strategy to enhance cancer cell apoptosis.