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Basic Research

Ethanol promotes cytotoxic effects of tumor necrosis factor-related apoptosis-inducing ligand through induction of reactive oxygen species in prostate cancer cells

Prostate Cancer and Prostatic Diseases volume 16, pages 16–22 (2013)Cite this article

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Abstract

Background:

Effective treatment of prostate cancer (PCa) remains a major challenge due to chemoresistance to drugs including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Ethanol and ethanol extracts are known apoptosis inducers. However, cytotoxic effects of ethanol on PCa cells are unclear.

Methods:

In this study we utilized PC3 and LNCaP cell culture models. We used immunohistochemical analysis, western blot analysis, reactive oxygen species (ROS) measurement, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) Cell Proliferation Assay, Annexin-V staining and flow cytometry for quantification of apoptosis. In vitro soft agar colony formation and Boyden chamber invasion assays were used. Tumorigenicity was measured in a xenotransplantation mouse model.

Results:

Here, we demonstrate that ethanol enhances the apoptosis-inducing potential of TRAIL in androgen-resistant PC3 cells and sensitizes TRAIL-resistant, androgen sensitive LNCaP cells to apoptosis through caspase activation, and a complete cleavage of poly (ADP)-ribose polymerase, which was in association with increased production of ROS. The cytotoxicity of ethanol was suppressed by an antioxidant N-acetyl cystein pretreatment. Furthermore, ethanol in combination with TRAIL increased the expression of cyclin-dependent kinase inhibitor p21 and decreased the levels of Bcl-2 and phosphorylated-AKT. These molecular changes were accompanied by decreased proliferation, anchorage-independent growth and invasive potential of PC3 and LNCaP cells. In vivo studies using a xenotransplantation mouse model with PC3 cells demonstrated significantly increased apoptosis in tumors treated with ethanol and TRAIL in combination.

Conclusions:

Taken together, use of ethanol in combination with TRAIL may be an effective strategy to augment sensitivity to TRAIL-induced apoptosis in PCa cells.

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Acknowledgements

This work was supported in part by Lake Champlain Cancer Research Organization (LCCRO) at the University of Vermont, Burlington, VT, Department of Neurosurgery, University of Wisconsin, Madison, WI, and NEAUA post residency research scholarship award. We thank Core Facilities of COBRE-Neuroscience (2P20RR16435-6) and Vermont Cancer Center (VCC) DNA analysis and Flow Cytometry Facility for their technical help.

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Authors and Affiliations

  1. Department of Surgery and Urology, University of Vermont, Burlington, VT, USA

    M K Plante & J B Folsom

  2. Vermont Cancer Center, University of Vermont, Burlington, VT, USA

    M K Plante, W T Arscott & S W Tighe

  3. Department of Neurosurgery, School of Medicine and Public Health, University of Wisconsin, Madison, WI, USA

    R J Dempsey & U V Wesley

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  1. M K Plante
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Correspondence to U V Wesley.

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Plante, M., Arscott, W., Folsom, J. et al. Ethanol promotes cytotoxic effects of tumor necrosis factor-related apoptosis-inducing ligand through induction of reactive oxygen species in prostate cancer cells. Prostate Cancer Prostatic Dis 16, 16–22 (2013). https://doi.org/10.1038/pcan.2012.37

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