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Retinoids induced astrocytic differentiation with down regulation of telomerase activity and enhanced sensitivity to taxol for apoptosis in human glioblastoma T98G and U87MG cells

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Abstract

We hypothesized that induction of differentiation with retinoid could increase sensitivity to microtubule-binding drug taxol (TXL) for apoptosis in human glioblastoma T98G and U87MG cells. 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, overexpression of glial fibrillary acidic protein (GFAP), and also down regulated telomerase expression and activity, thereby increased sensitivity to TXL for apoptosis. Treatment of glioblastoma cells with TXL triggered production of reactive oxygen species (ROS), induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and activated the redox-sensitive c-Jun NH2-terminal kinase 1 (JNK1) pathway. Moreover, TXL activated Raf-1 kinase for phosphorylation and inactivaion of anti-apoptotic Bcl-2 protein. The events of apoptosis included increase in expression of Bax, down regulation of Bcl-2 and baculoviral inhibitor-of-apoptosis protein (IAP) repeat containing (BIRC) proteins, mitochondrial release of cytochrome c and Smac into the cytosol, increase in intracellular free [Ca2+], and activation of calpain, caspase-9, and caspase-3. Increased activity of caspase-3 cleaved inhibitor of caspase-activated DNase (ICAD) to release and translocate CAD to the nucleus for DNA fragmentation. Involvement of stress signaling kinases and proteolytic activities of calpain and caspase-3 in apoptosis was confirmed by pretreating cells with specific inhibitors. Taken together, our results suggested that retinoid (ATRA or 13-CRA) induced astrocytic differentiation with down regulation of telomerase activity to increase sensitivity to TXL to enhance apoptosis in glioblastoma cells. Thus, combination of retinoid and TXL could be an effective therapeutic strategy for controlling the growth of glioblastoma.

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Abbreviations

ANOVA:

Analysis of variance

Asc:

Ascorbic acid

ATRA:

All-trans retinoic acid

BIRC:

Baculovirus IAP repeat containing

CAD:

Caspase-3-activated DNase

13-CRA:

13-cis Retinoic acid

DMSO:

Dimethyl sulfoxide

HRP:

Horseradish peroxidase

hTERT:

Human telomerase reverse transcriptase (hTERT)

IAP:

Inhibitor-of-apoptosis protein

ICAD:

Inhibitor of caspase-3-activated DNase

Kd :

Dissociation constant

MAPK:

Mitogen-activated protein kinase

p-NA:

p-Nitroanilide

OD:

Optical density

ROS:

Reactive oxygen species

RT-PCR:

Reverse transcription-polymerase chain reaction

SBDP:

Spectrin breakdown product

TXL:

Taxol

SB203580:

4-(4-Fluorophenyl)-2-(4-methylsulfinyl phenyl)-5-(4-pyridyl)-1H-imidazole

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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. Division of Neurology, Department of Neurosciences, Medical University of South Carolina (MUSC), 96 Jonathan Lucas Street, Suite 325E, P.O. Box 250606, Charleston, SC, 29425, USA

    Arabinda Das, Naren L. Banik & Swapan K. Ray

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

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Das, A., Banik, N.L. & Ray, S.K. Retinoids induced astrocytic differentiation with down regulation of telomerase activity and enhanced sensitivity to taxol for apoptosis in human glioblastoma T98G and U87MG cells. J Neurooncol 87, 9–22 (2008). https://doi.org/10.1007/s11060-007-9485-1

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