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Pretreatment with AT-101 enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis of breast cancer cells by inducing death receptors 4 and 5 protein levels

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Abstract

Purpose

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily and has been shown to induce extrinsic pathway of apoptosis in many types of cancer cells. AT-101, an (−)-enantiomer of gossypol, is a potent anticancer agent that is shown to be an inhibitor of Bcl-2/Bcl-XL. In this study, we searched whether pretreatment with either of these drugs would result in the enhancement of apoptosis through induction of death receptors and activation of mitochondrial pathways within breast cancer cells.

Methods

Human breast cancer (MCF-7 and MDA-MB-231) and normal breast cells (MCF-10A) were treated with drugs alone/in combination/sequentially. XTT cell viability assay was used to evaluate cytotoxicity. For showing apoptosis, both DNA Fragmentation and caspase 3/7 activity measurements were done. ELISA and Western blot analysis were done to assess DR4 and DR5 protein levels. The expression levels of apoptotic proteins were assessed by human apoptosis antibody array.

Results

The sequential treatment of AT-101 followed by TRAIL resulted in significant synergistic cytotoxicity and apoptosis. Moreover, pretreatment of breast cancer cells with AT-101 and then with TRAIL caused enhancement of the expression levels of DR4 and DR5 in both cancer cell lines, suggesting that these cells were under strong apoptotic stimuli.

Conclusions

These findings all together, strongly suggest that pretreatment with AT-101 enhances TRAIL-induced death-inducing signaling complex resulting in the engagement of the mitochondrial pathway to apoptosis in breast cancer cells. These promising, preliminary results make AT-101 and TRAIL a novel combination treatment candidate for breast cancer.

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Acknowledgments

This work was supported by grants from Celal Bayar University Scientific Research Projects Committee of Celal Bayar University. (Project No: FBE 2009-093).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Section of Molecular Biology, Department of Biology, Faculty of Science and Arts, Celal Bayar University, Muradiye, 45140, Manisa, Turkey

    Asli Kisim, Harika Atmaca & Selim Uzunoglu

  2. Division of Medical Oncology, Tulay Aktas Oncology Hospital, School of Medicine, Ege University, Bornova, 35100, Izmir, Turkey

    Burcu Cakar, Bulent Karabulut, Canfeza Sezgin, Ruchan Uslu & Burcak Karaca

Authors
  1. Asli Kisim
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  2. Harika Atmaca
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  3. Burcu Cakar
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  4. Bulent Karabulut
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  5. Canfeza Sezgin
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  6. Selim Uzunoglu
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  7. Ruchan Uslu
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  8. Burcak Karaca
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Correspondence to Burcak Karaca.

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Kisim, A., Atmaca, H., Cakar, B. et al. Pretreatment with AT-101 enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis of breast cancer cells by inducing death receptors 4 and 5 protein levels. J Cancer Res Clin Oncol 138, 1155–1163 (2012). https://doi.org/10.1007/s00432-012-1187-1

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  • DOI: https://doi.org/10.1007/s00432-012-1187-1

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