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Anti-proliferative Effects of Tricyclodecan-9-yl-xanthogenate (D609) Involve Ceramide and Cell Cycle Inhibition

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An Erratum to this article was published on 04 May 2012

Abstract

Tricyclodecan-9-yl-xanthogenate (D609) inhibits phosphatidylcholine (PC)-phospholipase C (PLC) and/or sphingomyelin (SM) synthase (SMS). Inhibiting SMS can increase ceramide levels, which can inhibit cell proliferation. Here, we examined how individual inflammatory and glia cell proliferation is altered by D609. Treatment with 100-μM D609 significantly attenuated the proliferation of RAW 264.7 macrophages, N9 and BV-2 microglia, and DITNC1 astrocytes, without affecting cell viability. D609 significantly inhibited BrdU incorporation in BV-2 microglia and caused accumulation of cells in G1 phase with decreased number of cells in the S phase. D609 treatment for 2 h significantly increased ceramide levels in BV-2 microglia, which, following a media change, returned to control levels 22 h later. This suggests that the effect of D609 may be mediated, at least in part, through ceramide increase via SMS inhibition. Western blots demonstrated that 2-h treatment of BV-2 microglia with D609 increased expression of the cyclin-dependent kinase (Cdk) inhibitor p21 and down-regulated phospho-retinoblastoma (Rb), both of which returned to basal levels 22 h after removal of D609. Exogenous C8-ceramide also inhibited BV-2 microglia proliferation without loss of viability and decreased BrdU incorporation, supporting the involvement of ceramide in D609-mediated cell cycle arrest. Our current data suggest that D609 may offer benefit after stroke (Adibhatla and Hatcher, Mol Neurobiol 41:206–217, 2010) through ceramide-mediated cell cycle arrest, thus restricting glial cell proliferation.

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Acknowledgements

Supported by NIH R01 NS063959 and AHA 11GRNT7360066 and resources provided by Veterans Affairs Hospital, Madison, WI. The authors have no conflict of interest to declare.

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Authors and Affiliations

  1. Department of Neurological Surgery, School of Medicine and Public Health, University of Wisconsin, Box 8660, Clinical Science Center, 600 Highland Ave, Madison, WI, 53792, USA

    Anchal Gusain, James F. Hatcher, Rao Muralikrishna Adibhatla, Umadevi V. Wesley & Robert J. Dempsey

  2. Cardiovascular Research Center, School of Medicine and Public Health, University of Wisconsin, Madison, WI, USA

    Rao Muralikrishna Adibhatla

  3. Neuroscience Training Program, School of Medicine and Public Health, University of Wisconsin, Madison, WI, USA

    Rao Muralikrishna Adibhatla

  4. William S. Middleton Veterans Affairs Hospital, Madison, WI, USA

    Anchal Gusain, James F. Hatcher, Rao Muralikrishna Adibhatla & Robert J. Dempsey

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  1. Anchal Gusain
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Correspondence to Rao Muralikrishna Adibhatla or Robert J. Dempsey.

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Anchal Gusain and James F. Hatcher contributed equally to this manuscript.

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Gusain, A., Hatcher, J.F., Adibhatla, R.M. et al. Anti-proliferative Effects of Tricyclodecan-9-yl-xanthogenate (D609) Involve Ceramide and Cell Cycle Inhibition. Mol Neurobiol 45, 455–464 (2012). https://doi.org/10.1007/s12035-012-8254-0

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