Attenuated Leishmanial sphingolipid induces apoptosis in A375 human melanoma cell via both caspase-dependent and -independent pathways

  • Jagnyeswar Ratha
  • Kajal Nayan Majumdar
  • Koushik Dhara
  • Suman Kumar Singh
  • Krishna Das Saha
  • Ranjan Bhadra


A fraction of attenuated Leishmanial lipid (ALL) rich in sphingolipids, previously shown to have apoptosis inducing activity in mouse melanoma (B16F10) and human melanoma (A375) cells, was resolved to isolate the bioactive sphingolipid. The mechanism of apoptosis induction by this bioactive attenuated Leishmanial sphingolipid (ALSL) was studied in A375 cells. Apoptosis induced by ALSL in A375 cells was found to be dose and time-dependent. Exposure of cells to ALSL resulted in a rapid increase in reactive oxygen species generation. Pretreatment of cells with the antioxidant N-acetyl-cystein reduced ROS generation and attenuated apoptosis induced by ALSL. Again, ALSL sensitization resulted in the activation of caspase-3 and -9 but not caspase-8. However, inhibitors of these caspases could not protect the cells completely from ALSL-induced apoptosis. N-acetyl-cystein pretreatment was again found to attenuate the activation of caspase-3 and -9. ALSL treatment also resulted in the alteration of mitochondrial membrane potential, and release of pro-apoptotic factors such as cytochrome c and apoptosis inducing factor (AIF) from mitochondria. Furthermore, c-Jun N-terminal kinase was activated that resulted in apoptosis of A375 cells, whereas p38 MAPK was activated to counteract the stress generated in cells in response to ALSL treatment. Taken together, our results indicate that ALSL-induced apoptosis of A375 cells is mediated by both mitochondrial caspase-dependent and -independent pathways and it involves ROS and JNK activation in the mitogen-activated protein kinase cascade.


Attenuated Leishmanial sphingolipid Apoptosis ROS AIF Melanoma 



We thank our director, Prof. Siddhartha Roy for his patronization of and keen interest in this work. We are indebted to Dr. Basudev Achari of our institute for critically reviewing the manuscript. Financial assistance from the Council of Scientific and Industrial Research (CSIR), India is thankfully acknowledged.


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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Jagnyeswar Ratha
    • 1
  • Kajal Nayan Majumdar
    • 1
  • Koushik Dhara
    • 2
  • Suman Kumar Singh
    • 3
  • Krishna Das Saha
    • 1
  • Ranjan Bhadra
    • 4
  1. 1.Cellular Biochemistry DivisionIndian Institute of Chemical BiologyKolkataIndia
  2. 2.Department of Inorganic ChemistryIndian Association for the Cultivation of ScienceKolkataIndia
  3. 3.Department of Biomedical SciencesUniversity of BradfordBradford, West YorkshireUK
  4. 4.Vijaygarh Jyotish Ray CollegeResearch & Development CenterKolkataIndia

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