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Molecular Mechanisms of the Combination of Retinoid and Interferon-gamma for Inducing Differentiation and Increasing Apoptosis in Human Glioblastoma T98G and U87MG Cells

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Abstract

Glioblastoma is the deadliest brain tumor that remains incurable. We examined efficacy of combination of retinoid and interferon-gamma (IFN-γ) in human glioblastoma T98G and U87MG cells. We conjectured that retinoid could induce differentiation with down regulation of telomerase activity to increase sensitivity to IFN-γ for apoptosis in glioblastoma cells. Indeed, treatment of cells with 1 μM all-trans retinoic acid (ATRA) or 1 μM 13-cis retinoic acid (13-CRA) for 7 days induced astrocytic differentiation with upregulation of glial fibrillary acidic protein (GFAP) and down regulation of telomerase activity. Wright staining and ApopTag assay showed, respectively, morphological and biochemical features of apoptosis in glioblastoma cells following exposure to 200 units/ml IFN-γ for 48 h. Induction of differentiation was associated with decreases in levels of nuclear factor kappa B (NFκB), inducible nitric oxide synthase (iNOS), and production of nitric oxide (NO) so as to increase sensitivity to IFN-γ for apoptosis. Notably, IFN-γ induced signal transducer and activator of transcription-1 (STAT-1) to bind to gamma-activated sequence (GAS) of the target gene. Also, IFN-γ activated caspase-8 and cleaved Bid to truncated Bid (tBid) for translocation to mitochondria. Fura-2 assay showed increases in intracellular free [Ca2+] and activation of calpain in apoptotic cells. Besides, increases in Bax:Bcl-2 ratio and mitochondrial release of cytochrome c and Smac into the cytosol activated caspase-9 and caspase-3 for apoptosis. Taken together, our results showed that retinoid induced astrocytic differentiation with down regulation of telomerase activity and enhanced sensitivity to IFN-γ for increasing apoptosis in human glioblastoma cells.

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Acknowledgments

This investigation was supported in part by the R01 grants (CA-91460 and NS-57811) from the National Institutes of Health (Bethesda, MD, USA).

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Authors and Affiliations

  1. Department of Neurosciences, Medical University of South Carolina, Charleston, SC, 29425, USA

    Arabinda Das & Naren L. Banik

  2. Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, 6439 Garners Ferry Road, Columbia, SC, 29209, USA

    Swapan K. Ray

Authors
  1. Arabinda Das
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  2. Naren L. Banik
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  3. Swapan K. Ray
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Correspondence to Swapan K. Ray.

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Special issue article in honor of Dr. George DeVries.

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Das, A., Banik, N.L. & Ray, S.K. Molecular Mechanisms of the Combination of Retinoid and Interferon-gamma for Inducing Differentiation and Increasing Apoptosis in Human Glioblastoma T98G and U87MG Cells. Neurochem Res 34, 87–101 (2009). https://doi.org/10.1007/s11064-008-9669-x

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  • DOI: https://doi.org/10.1007/s11064-008-9669-x

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