Molecular signatures of N-nitroso compounds in Caco-2 cells: implications for colon carcinogenesis.
N-nitroso compounds (NOC) are genotoxic, carcinogenic to animals, and may play a role in human cancer development. Because the gastro-intestinal tract is an important route of exposure through endogenous nitrosation, we hypothesize that NOC exposure targets genetic processes relevant in colon carcinogenesis. To investigate these genomic responses, we analyzed the transcriptomic effects of genotoxic concentrations of two nitrosamides, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, 1 microM) and N-methyl-N-nitrosurea (MNU, 1 mM), and four nitrosamines, N-nitrosodiethylamine (NDEA, 50mM), N-nitrosodimethylamine (NDMA, 100 mM), N-nitrosopiperidine (NPIP, 40 mM), and N-nitrosopyrrolidine (NPYR, 100mM), in the human colon carcinoma cell line Caco-2. Gene Ontology gene group, consensus motif gene group and biological pathway analysis revealed that nitrosamides had little effect on gene expression after 24 h of exposure, whereas nitrosamines had a strong impact on the transcriptomic profile. Analyses showed modifications of cell cycle regulation and apoptosis pathways for nitrosamines which was supported by flow cytometric analysis. We found additional modifications in gene groups and pathways of oxidative stress and inflammation, which suggest an increase in oxidative stress and proinflammatory immune response upon nitrosamine exposure, although less distinct for NDMA. Furthermore, NDEA, NPIP, and NPYR most strongly affected several developmental motif gene groups and pathways, which may influence developmental processes. Many of these pathways and gene groups are implicated in the carcinogenic process and their modulation by nitrosamine exposure may therefore influence the development of colon cancer. In summary, our study has identified pathway modifications in human colon cells which may be associated with cancer risk of nitrosamine exposure in the human colon.
Hebels
Dennie G A J
DG
Department of Health Risk Analysis and Toxicology, Maastricht University, 6200 MD Maastricht, The Netherlands. d.hebels@grat.unimaas.nl
Jennen
Danyel G J
DG
Kleinjans
Jos C S
JC
de Kok
Theo M C M
TM
D017209:Apoptosis / Q000187:drug effects; D018938:Caco-2 Cells; D002453:Cell Cycle / Q000187:drug effects; D003110:Colonic Neoplasms / Q000139:chemically induced* / Q000473:pathology*; D020552:Comet Assay; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005819:Genetic Markers; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009603:Nitroso Compounds / Q000737:chemistry* / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification
D005819:Genetic Markers; D009603:Nitroso Compounds; D012313:RNA