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Enhanced 5-fluorouracil cytotoxicity in high cyclooxygenase-2 expressing colorectal cancer cells and xenografts induced by non-steroidal anti-inflammatory drugs via downregulation of dihydropyrimidine dehydrogenase

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Abstract

Purpose

To prove that 5-FU cytotoxicity could be increased by combination with low-dose non-steroidal anti-inflammatory drugs (NSAIDs) (indomethacin or NS-398) in high cyclooxygenase-2- (COX-2) expressing cells and xenografts through the modulation of dihydropyrimidine dehydrogenase (DPD) mRNA expression and/or enzyme activity.

Methods

HT-29 cells were grown on collagen IV coated plates (HT-29-C). The antiproliferative effect of 5-fluorouracil (5-FU) ± NSAIDs was examined on non-COX-2 expressing HT-29 and COX-2-expressing HT-29-C cells by sulphorhodamine B assay. The COX-2 and DPD expressions were visualized by immunofluorescent staining, and prostaglandin E2 levels were measured by ELISA kit. The HT-29 xenograft was established in SCID mice and treated with 5-FU ± NSAIDs for 5 days. The tumor volume, enzyme activity, and DPD mRNA expression were investigated by caliper, radioenzymatic method, and real-time RT-PCR, respectively. The drug interaction was calculated for both combinations (5-FU + indomethacin and 5-FU + NS-398).

Results

Collagen IV up-regulated significantly the COX-2 and DPD mRNA, and protein expressions, and also their enzyme activities in HT-29 cells. NSAIDs enhanced in a synergistic manner the cytotoxic effect of 5-FU treatment both in vitro and in vivo. Downregulation of DPD was observed after 5-FU monotherapy, but the combined effect of NSAIDs and 5-FU on DPD mRNA expression, and enzyme activity was superior to the effect of 5-FU alone.

Conclusions

Since 5-FU + NSAID treatment can alter the DPD enzyme activity resulting in an enhanced cytotoxic effect, further studies in clinical practice are warranted.

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Acknowledgments

The SCID mice were generously gifted by Dr. D. Gaál. The excellent technical assistance of Cs. Polényi Makácsné, A. Nagy, J. Kútvölgyi, A. Éber Mousáné, A. Sztodola and A. M. Borza are greatly appreciated. This study was supported by the Jedlik Ányos Grant (NKFP1-00024/2005).

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Authors and Affiliations

  1. Department of Clinical Research, National Institute of Oncology, Budapest, Hungary

    Andrea Réti, Éva Pap, Vilmos Adleff, Judit Kralovánszky & Barna Budai

  2. 1st Institute of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary

    András Jeney

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  1. Andrea Réti
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Correspondence to Judit Kralovánszky.

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Réti, A., Pap, É., Adleff, V. et al. Enhanced 5-fluorouracil cytotoxicity in high cyclooxygenase-2 expressing colorectal cancer cells and xenografts induced by non-steroidal anti-inflammatory drugs via downregulation of dihydropyrimidine dehydrogenase. Cancer Chemother Pharmacol 66, 219–227 (2010). https://doi.org/10.1007/s00280-009-1149-8

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