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The apoptotic pathway triggered by the Fhit protein in lung cancer cell lines is not affected by Bcl-2 or Bcl-x(L) overexpression

Oncogene volume 23, pages 9102–9110 (2004)Cite this article

Abstract

The expression of the tumour suppressor protein fragile histidine triad (Fhit) is often impaired in many human cancers and its restoration in Fhit-negative cancer cell lines suppresses tumorigenicity and induces apoptosis. Although the proapoptotic function of Fhit is well documented, little is known about its precise mechanism of action and further studies are needed in order to elucidate the putative therapeutic properties of this protein. To this end, we have engineered the lung cancer cell line NCI-H460 in order to express different molecules involved in the control of apoptotic pathways. Infection of these cells with an adenoviral vector transducing the Fhit gene (Ad-Fhit) revealed that complete protection from apoptosis was conferred by the inhibitor of caspases Cytokine response modifier A (CrmA) and by a dominant-negative form of the adapter protein Fas-associated death domain (FADD) and partial protection by a dominant-negative form of caspase-8, while cells over expressing mitochondrial mediators of the apoptotic response such as Bcl-2 or Bcl-x(L) that are resistant to treatment with cisplatin, remained highly susceptible to cell death triggered by Fhit gene transfer. In line to what was observed in H460 cells, Ad-Fhit efficacy was not affected by Bcl-2 overexpression also in two other lung cancer cell lines (A549 and Calu-1). Analysis of cytochrome c release also confirmed that in Bcl-2- or Bcl-x(L)-expressing cells apoptosis could be detected by terminal deoxynucleotidyl-transferase mediated dUTP nick-end labelling (TUNEL) assay before any evidence of mitochondrial membrane perturbation. In conclusion, our analysis indicates that the Fhit protein exerts its oncosuppressor activity through induction of an apoptotic mechanism that seems to be FADD dependent, caspase-8 mediated and independent from mitochondrial amplification.

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Abbreviations

Fhit:

fragile histidine triad

NSLC:

non-small-cell lung cancer

FADD:

Fas-associated death domain

CrmA:

cytokine response modifier A

MOI:

multiplicity of infection

TUNEL:

terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling

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Acknowledgements

We thank Professor P Krammer (Deutsches Krebsforschungszentrum, Heidelberg, Germany) for the anti-caspase-8 antibody, Drs Massimo Zeviani and Valeria Tiranti (Istituto Nazionale Neurologico ‘C Besta’, Milan, Italy) for the anti-COX IV antibody and Drs Delia Mezzanzanica and Paola Perego (Istituto Nazionale Tumori, Milan, Italy) for helpful discussions and for providing the anti-cytochrome c and anti-Bcl-X(L) antibodies, respectively. We are also grateful to Dr Hideshi Ishii for technical advice and Professor Carlo M Croce (both at Kimmel Cancer Center, Jefferson Medical College, Philadelphia, PA, USA) for comments and suggestions. This work was partially supported by grants from Associazione Italiana per la Ricerca sul Cancro (AIRC) and from Ministero Italiano della Sanita' to GS. LR and FA are recipients of fellowships from AIRC and Ministero Italiano della Sanita'.

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Author notes

  1. Carlos G Ferreira

    Present address: Clinical and Translational Research Service, Coordenaçäo de Pesquisa, Instituto Nacional de Cancer (INCA), Rio De Janeiro, RJ, Brazil

Authors and Affiliations

  1. Department of Experimental Oncology, Istituto Nazionale Tumori, Via Venezian 1, Milan, 20133, Italy

    Luca Roz, Francesca Andriani & Gabriella Sozzi

  2. Department of Medical Oncology, Vrije Universiteit Medical Center, Amsterdam, HV 1081, The Netherlands

    Carlos G Ferreira & Giuseppe Giaccone

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  1. Luca Roz
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  2. Francesca Andriani
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  3. Carlos G Ferreira
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  5. Gabriella Sozzi
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Correspondence to Luca Roz.

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Roz, L., Andriani, F., Ferreira, C. et al. The apoptotic pathway triggered by the Fhit protein in lung cancer cell lines is not affected by Bcl-2 or Bcl-x(L) overexpression. Oncogene 23, 9102–9110 (2004). https://doi.org/10.1038/sj.onc.1208142

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