title: The effect of clotrimazole on energy substrate uptake and carcinogenesis in intestinal epithelial cells.
abstract: Clotrimazole has anticarcinogenic activity in several cell types. Our aims were to investigate the anticarcinogenic effect of clotrimazole in a tumoral intestinal epithelial (Caco-2) cell line, to compare it with the effect in a nontumoral intestinal epithelial cell line (IEC-6 cells), and to investigate inhibition of energy substrate uptake as a mechanism contributing to it. The effect of clotrimazole on cell proliferation, viability and differentiation, H-deoxyglucose (H-DG), H-O-methyl-glucose (H-OMG), and C-butyrate uptake, as well as mRNA expression levels of glucose transporters was assessed. In Caco-2 cells, clotrimazole decreased cellular viability and proliferation and increased cell differentiation. The effect on cell proliferation and viability was potentiated by rhodamine123. Clotrimazole also decreased cellular viability and proliferation in IEC-6 cells, but increased the cellular DNA synthesis rate and had no effect on cell differentiation. Exposure of Caco-2 cells to clotrimazole (10 µmol/l) for 1 and 7 days increased (by 20-30%) the uptake of H-DG and H-OMG, respectively, but had no effect on C-butyrate uptake. The effect on H-DG and H-OMG transport was maximal at 10 µmol/l, and the pharmacological characteristics of transport were not changed. However, clotrimazole changed the mRNA expression levels of the facilitative glucose transporter 2 and the Na-dependent glucose cotransporter. Clotrimazole exhibits comparable cytotoxic effects in tumoral and nontumoral intestinal epithelial cell lines. In Caco-2 cells, the cytotoxic effect of clotrimazole was strongly potentiated by the inhibition of oxidative phosphorylation. Moreover, stimulation of glucose uptake might be a compensation mechanism in response to the glycolysis inhibition caused by clotrimazole.
authors.affiliation: Department of Biochemistry (U38-FCT), University of Porto, Porto, Portugal.
authors.forename: Pedro
authors.initials: P
authors.lastname: Gonçalves
authors.forename: Inês
authors.initials: I
authors.lastname: Gregório
authors.forename: João R
authors.initials: JR
authors.lastname: Araújo
authors.forename: Fátima
authors.initials: F
authors.lastname: Martel
mesh_terms: D019325:3-O-Methylglucose / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002087:Butyrates / Q000378:metabolism; D018938:Caco-2 Cells; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003022:Clotrimazole / Q000494:pharmacology*; D004247:DNA / Q000096:biosynthesis; D003847:Deoxyglucose / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051246:Glucose Transport Proteins, Facilitative / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D010085:Oxidative Phosphorylation; D020112:Rhodamine 123 / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D002087:Butyrates; D051246:Glucose Transport Proteins, Facilitative; D019325:3-O-Methylglucose; D020112:Rhodamine 123; D004247:DNA; D003847:Deoxyglucose; D003022:Clotrimazole