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The apoptosis-inducing effect of gastrin on colorectal cancer cells relates to an increased IEX-1 expression mediating NF-κB inhibition

Abstract

Addressing the puzzling role of amidated gastrin17 (G17) and the gastrin/CCKB/CCK2 receptor in colorectal carcinogenesis, we analysed potential candidate genes involved in G17-dependent NF-κB inhibition and apoptosis. The colorectal carcinoma cell line Colo320 overexpressing the wild-type CCK2 receptor (Colo320wt) underwent G17-induced apoptosis along with suppressed NF-κB activation and decreased expression of the antiapoptotic NF-κB target genes cIAP1 and cIAP2, whereas G17 was without effect on Colo320 cells expressing a CCK2 receptor bearing a loss of function mutation (Colo320mut). Gene microarray analysis revealed an elevated expression of the stress response gene IEX-1 in G17-treated Colo320wt but not Colo320mut cells. Quantitative real-time PCR and conventional RT–PCR confirmed this G17-dependent increase of IEX-1 expression in Colo320wt cells. If these cells were subjected to IEX-1 knockdown by small interfering RNA transfection, the apoptosis-inducing effect of G17 was abolished. Moreover, tumor necrosis factor alpha (TNFα)- or 5-FU-induced apoptosis that is greatly enhanced by G17 treatment in Colo320wt cells was prevented if IEX-1 expression was repressed. Under these conditions of blocked IEX-1 expression, the NF-κB activity remained unaffected by G17, in particular in Colo320wt cells co-treated with TNFα and also the suppressive effect of G17 on cIAP1 and cIAP2 expression was not observed anymore if IEX-1 expression was blocked. Conversely, IEX-1 overexpression in Colo320mut cells caused an increase of basal and TNFα- or 5-FU-induced apoptosis, an effect not further triggered by G17 treatment. Using a xenograft tumor model in severe combined immune deficiency mice, we could show that experimental systemic hypergastrinemia induced by the administration of omeprazole led to enhanced apoptosis as well as to a marked increase of IEX-1 expression in Colo320wt tumors, but not in Colo320mut tumors. These observations indicate that the proapoptotic effect of G17 on human colon cancer cells expressing the wild-type CCK2 receptor is mediated by IEX-1, which modulates NF-κB-dependent antiapoptotic protection and thereby exerts tumor-suppressive potential.

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Abbreviations

CCK:

cholecystokinin

cIAP1/2:

cellular inhibitor of apoptosis protein 1/2

5-FU:

5-fluoro uracil

G17:

amidated gastrin (17–34)

IEX-1:

immediate-early gene on X-ray-1

MAPK:

mitogen-activated protein kinase

NF-κB:

nuclear factor kappa B

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Acknowledgements

This work was supported by a grant of the German Research Society DFG (SFB415/A13 to HS) and the Medical Faculty of the Kiel University (to SSM).

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Correspondence to H Schäfer.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Sebens Müerköster, S., Rausch, A., Isberner, A. et al. The apoptosis-inducing effect of gastrin on colorectal cancer cells relates to an increased IEX-1 expression mediating NF-κB inhibition. Oncogene 27, 1122–1134 (2008). https://doi.org/10.1038/sj.onc.1210728

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