Interferon-γ-Induced Sensitization of Colon Carcinomas to ZD9331 Targets Caspases, Downstream of Fas, Independent of Mitochondrial Signaling and the Inhibitor of Apoptosis Survivin

James Geller, I. Peták, Kinga Szekely Szucs, Katalin Nagy, David M. Tillman, Janet A. Houghton

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Abstract

We have demonstrated previously a Fas-dependent component in thymineless death of human colon carcinoma cells. Importantly, the cytotoxic effects of thymidine deprivation induced by 5-fluorouracil (FUra) combined with leucovorin (LV) was enhanced by IFN-γ, and the synergism was shown to be dependent on Fas, FUra-induced DNA damage, and independent of p53. Subsequently we examined the potential for synergistic interactions between IFN-γ and the specific thymidylate synthase inhibitor, ZD9331. IFN-γ sensitized colon carcinomas to ZD9331-induced apoptosis and loss in clonogenic survival, also dependent on ZD9331-induced DNA damage, independent of p53. Synergism occurred in HCT116, demonstrating previously RNA-mediated FUra/LV cytotoxicity that could not be potentiated by IFN-γ. Manipulation of the Fas death receptor pathway from the level of the receptor (Nok1/Nok2, Fas overexpression, and DN-FADD) to the mitochondria (Bcl-xL and Bcl-2) did not modulate ZD9331 ± IFN-γ-induced cytotoxicity in HT29, with the exception that Nok1/Nok2-blocking antibodies partially protected HT29 from the cytotoxic activity of ZD9331 alone. However, IFN-γ alone (but not ZD9331) up-regulated the expression of caspases -3, -4, -7, and -8, and in combination with ZD9331 demonstrated enhanced caspase activation and cleavage of poly(ADP-ribose) polymerase that was not prevented by overexpression of Bcl-2. Additionally, IFN-γ increased the activity of the proteasome in HT29, leading to selective down-regulation of the anti-apoptotic protein survivin, whereas simultaneously increasing Fas expression. However, reduction in the survivin:Fas ratio by transfection of survivin small interfering RNA and/or overexpression of Fas did not affect sensitivity of HT29 to ZD9331 ± IFN-γ. Data demonstrate that IFN-γ combined with ZD9331 is synergistic in additional cell lines that demonstrate RNA-mediated FUra/LV cytotoxicity, and that a major target of interaction is at the level of caspases, downstream of Fas, and independent of involvement of either the mitochondria or survivin.

Original languageEnglish
Pages (from-to)6504-6515
Number of pages12
JournalClinical Cancer Research
Volume9
Issue number17
Publication statusPublished - Dec 15 2003

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Caspases
Interferons
Colon
Apoptosis
Carcinoma
Fluorouracil
Leucovorin
DNA Damage
Mitochondria
RNA
CD95 Antigens
ZD 9331
Thymidylate Synthase
Death Domain Receptors
Apoptosis Regulatory Proteins
Blocking Antibodies
Poly(ADP-ribose) Polymerases
Proteasome Endopeptidase Complex
Caspase 3
Thymidine

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Interferon-γ-Induced Sensitization of Colon Carcinomas to ZD9331 Targets Caspases, Downstream of Fas, Independent of Mitochondrial Signaling and the Inhibitor of Apoptosis Survivin. / Geller, James; Peták, I.; Szucs, Kinga Szekely; Nagy, Katalin; Tillman, David M.; Houghton, Janet A.

In: Clinical Cancer Research, Vol. 9, No. 17, 15.12.2003, p. 6504-6515.

Research output: Contribution to journalArticle

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