In vitro and in vivo evaluation of NCX 4040 cytotoxic activity in human colon cancer cell lines

Anna Tesei¹ , Paola Ulivi² , Francesco Fabbri¹ , Marco Rosetti² , Carlo Leonetti³ , Marco Scarsella³ , Gabriella Zupi³ , Dino Amadori¹ , Manlio Bolla⁴ and Wainer Zoli¹

¹Division of Oncology and Diagnostics, Morgagni-Pierantoni Hospital, Forlì, Italy

²Istituto Oncologico Romagnolo, Forlì, Italy

³Preclinical Experimental Laboratory, Regina Elena Institute for Cancer Research, Rome, Italy

⁴NicOx SA, Sophia-Antipolis, France

author email corresponding author email

Journal of Translational Medicine 2005, 3:7doi:10.1186/1479-5876-3-7


Published:	3 February 2005

Abstract

Background

Nitric oxide-releasing nonsteroidal antiinflammatory drugs (NO-NSAIDs) are reported to be safer than NSAIDs because of their lower gastric toxicity. We compared the effect of a novel NO-releasing derivate, NCX 4040, with that of aspirin and its denitrated analog, NCX 4042, in in vitro and in vivo human colon cancer models and investigated the mechanisms of action underlying its antitumor activity.

Methods

In vitro cytotoxicity was evaluated on a panel of colon cancer lines (LoVo, LoVo Dx, WiDr and LRWZ) by sulforhodamine B assay. Cell cycle perturbations and apoptosis were evaluated by flow cytometry. Protein expression was detected by Western blot. In the in vivo experiments, tumor-bearing mice were treated with NCX 4040, five times a week, for six consecutive weeks.

Results

In the in vitro studies, aspirin and NCX 4042 did not induce an effect on any of the cell lines, whereas NCX 4040 produced a marked cytostatic dose-related effect, indicating a pivotal role of the -NO₂group. Furthermore, in LoVo and LRWZ cell lines, we observed caspase-9 and -3-mediated apoptosis, whereas no apoptotic effect was observed after drug exposure in WiDr or LoVo Dx cell lines. In in vivo studies, both NCX 4040 and its parental compound were administered per os. NCX 4040 induced a 40% reduction in tumor weight. Conversely, aspirin did not influence tumor growth at all.

Conclusions

NCX 4040, but not its parental compound, aspirin, showed an in vitro and in vivo antiproliferative activity, indicating its potential usefulness to treat colon cancer.