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Autophagy modulators sensitize prostate epithelial cancer cell lines to TNF-alpha-dependent apoptosis

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Abstract

TNF-alpha levels in prostate cancer correlate with the extent of disease and are significantly elevated in the metastatic stage. TNF receptor superfamily controls two distinct signalling cascades, leading to opposite effects, i.e. apoptosis and survival; in prostate cancer TNF-alpha-mediated signalling induces cell survival and resistance to therapy. The apoptosis of prostate epithelial cancer cells LNCaP and PC3 was investigated upon treatment with the autophagy inhibitor 3-methyladenine and the autophagy inducer rapamycin, in combination with TNF-alpha. Cells were exposed to these molecules for 18, 24 and 48 h. Autophagy was assessed via LC3 Western blot analysis; propidium iodide and TUNEL stainings followed by flow cytometry or caspase-8 and caspase-3 activation assays were performed to evaluate apoptosis. TNF-alpha-induced apoptosis was potentiated by 3-methyladenine in the androgen-responsive LNCaP cells, whereas no effect was observed in the androgen-insensitive PC3 cells. Interestingly such pro-apoptosis effect in LNCaP cells was associated with reduced c-Flip levels through proteasomal degradation via increased reactive oxygen species production and p38 activation; such c-Flip reduction was reversed in the presence of either the proteasome inhibitor MG132 or the reactive oxygen species scavenger N-acetyl-cysteine. Conversely in PC3 but not in LNCaP cells, rapamycin stimulated TNF-alpha-dependent apoptosis; such effect was associated with reduced c-Flip promoter activity and FoxO3a activation. We conclude that TNF-alpha-induced apoptosis may be potentiated, in prostate cancer epithelial cells, through autophagy modulators. Increased sensitivity to TNF-alpha-dependent apoptosis correlates with reduced c-Flip levels which are consequent to a post-transcriptional and a transcriptional mechanism in LNCaP and PC3 cells respectively.

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Acknowledgments

We thank Dr Francesco Facchiano, Dr Stefania Rossi (Istituto Superiore di Sanità, Rome, Italy) and Dr Pierpaolo Coluccia (Sapienza University, Rome, Italy) for technical support. Fondazione Roma (http://www.fondazioneroma.it) and International FIRB (Fondo per gli Investimenti della Ricerca di Base Internazionale) (http://www.istitutopasteur.it) co-funded this work. The Authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. Department of Anatomy, Histology, Forensic Medicine and Orthopedics-Section of Histology and Medical Embryology, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via A. Scarpa, 14, 00161, Rome, Italy

    Claudia Giampietri, Simonetta Petrungaro, Fabrizio Padula, Elettra Sara Marini, Antonio Filippini & Elio Ziparo

  2. School of Medicine, Institute of Histology and Embryology, Catholic University of the Sacred Heart, 00168, Rome, Italy

    Alessio D’Alessio

  3. Laboratory of Vascular Pathology, Istituto Dermopatico dell’Immacolata, IDI-IRCCS, 00167, Rome, Italy

    Antonio Facchiano

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  1. Claudia Giampietri
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  2. Simonetta Petrungaro
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  3. Fabrizio Padula
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  4. Alessio D’Alessio
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Correspondence to Claudia Giampietri.

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Giampietri, C., Petrungaro, S., Padula, F. et al. Autophagy modulators sensitize prostate epithelial cancer cell lines to TNF-alpha-dependent apoptosis. Apoptosis 17, 1210–1222 (2012). https://doi.org/10.1007/s10495-012-0752-z

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