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Rapid and profound potentiation of Apo2L/TRAIL-mediated cytotoxicity and apoptosis in thoracic cancer cells by the histone deacetylase inhibitor Trichostatin A: the essential role of the mitochondria-mediated caspase activation cascade

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Abstract

Apo2L/TRAIL is actively investigated as a novel targeted agent to directly induce apoptosis of susceptible cancer cells. Apo2L/TRAIL-refractory cells can be sensitized to the cytotoxic effect of this ligand by cytotoxic chemotherapeutics. The aim of this study was to evaluate the in vitro tumoricidal activity of the Apo2L/TRAIL + Trichostatin A in cultured thoracic cancer cells and to elucidate the molecular basis of the synergistic cytotoxicity of this combination. Concurrent exposure of cultured cancer cells to sublethal concentrations of Apo2L/TRAIL and Trichostatin A resulted in profound enhancement of Apo2L/TRAIL-mediated cytotoxicity in all cell lines regardless of their intrinsic susceptibility to this ligand. This combination was not toxic to primary normal cells. While Apo2L/TRAIL alone or Trichostatin A alone mediated < 20% cell death, 60 to 90% of cancer cells were apoptotic following treatment with TSA + Apo2L/TRAIL combinations. Complete translocation of Bax from the cytosol to the mitochondria compartment was mainly observed in combination-treated cells and this was correlated with robust elevation of caspase 9 proteolytic activity indicative of activation of the mitochondria apoptogenic effect. Profound TSA + Apo2L/TRAIL–mediated cytotoxicity and apoptosis were completely abrogated by either Bcl2 over-expression or by the selective caspase 9 inhibitor, highlighting the essential role of mitochondria-dependent apoptosis signaling cascade in this process. Moreover, increased caspase 8 activity observed in cells treated with the TSA + Apo2L/TRAIL combination was completely suppressed by Bcl-2 over-expression or by the selective caspase 9 inhibitor indicating that the elevated caspase 8 activity in combination-treated cells was secondary to a mitochondria-mediated amplification feedback loop of caspase activation. These finding form the basis for further development of HDAC inhibitors + Apo2L/TRAIL combination as novel targeted therapy for thoracic malignancies.

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Abbreviations

TSA:

Trichostatin A

TRAIL:

TNF-related apoptosis inducing ligand

NSCLC:

non-small cell lung cancer

EsC:

esophageal cancer

MPM:

malignant pleural mesothelioma

HDACI:

Histone deacetylase inhibitor

TUNEL:

terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling

MTT:

(4,5-dimethylthiazo-2-yl)-2,5-diphenyl tetrazolium bromide

Apaf-1:

apoptotic protease activating factor 1

SMAC/DIABLO:

second mitochondria-derived activator

AIF:

apoptosis inducing factor

GFP:

green fluorescence protein

RIP:

receptor interacting protein

FLIP:

FLICE inhibitory protein

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Authors and Affiliations

  1. Section of Thoracic Oncology, Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Rishindra M. Reddy, Wen-Shuz Yeow, Alex Chua, Duc M. Nguyen, Aris Baras, M. Firdos Ziauddin, Susan M. Shamimi-Noori, Justin B. Maxhimer, David S. Schrump & Dao M. Nguyen

  2. Research Scholar, Howard Hughes Medical Institute, Bethesda

    Susan M. Shamimi-Noori & Justin B. Maxhimer

  3. Recipients of Cancer Research Training Award, NCI, NIH, MD

    Alex Chua, Duc M. Nguyen & Aris Baras

  4. Clinical Research Center, Room 4W-4-3940, 10 Center Drive, Bethesda, MD, 20892-1201, USA

    Dao M. Nguyen

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  1. Rishindra M. Reddy
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  2. Wen-Shuz Yeow
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  3. Alex Chua
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  4. Duc M. Nguyen
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  5. Aris Baras
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  6. M. Firdos Ziauddin
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  7. Susan M. Shamimi-Noori
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  8. Justin B. Maxhimer
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  9. David S. Schrump
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  10. Dao M. Nguyen
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Correspondence to Dao M. Nguyen.

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R.M. Reddy and W.-S. Yeow contributed equally to this work.

This research was supported by the Intramural Research Program of the National Cancer Institute, NIH.

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Reddy, R.M., Yeow, WS., Chua, A. et al. Rapid and profound potentiation of Apo2L/TRAIL-mediated cytotoxicity and apoptosis in thoracic cancer cells by the histone deacetylase inhibitor Trichostatin A: the essential role of the mitochondria-mediated caspase activation cascade. Apoptosis 12, 55–71 (2007). https://doi.org/10.1007/s10495-006-0484-z

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