Butyrate induces glutathione S-transferase in human colon cells and protects from genetic damage by 4-hydroxy-2-nonenal.
Butyrate, one of the major products of gut fermentation, is known to inhibit proliferation, induce apoptosis and differentiation, and increase phase II enzyme activities in tumor cells, whereas little information is available on protective effects in less-transformed colon cells. The aim of this study was to investigate whether the chemoprotective mechanism of glutathione S-transferase (GST) induction by butyrate could also play a role in earlier stages of colon carcinogenesis and whether chemoresistance of cells toward the endogenous genotoxic risk factor 4-hydroxy-2-nonenal (HNE) could be a consequence of butyrate treatment. As cell models, we used the human tumor cell lines HT29 and HT29 clone 19A, a differentiated subclone with properties resembling primary colon cells. We determined the expression of GSTP1 protein (enzyme-linked immunosorbent assay), the major GST in HT29, GSTP1 mRNA (Northern blotting), GST activity, intracellular glutathione, and total protein. The genotoxic impact of HNE (100-200 microM) was compared in butyrate-treated and nontreated cells using single-cell microgel electrophoresis. Our results show that GSTP1 mRNA, GSTP1 protein, GST activity, and total protein were increased (1.2- to 2.5-fold) and glutathione levels were maintained after 24-72 h of incubation with 4 mM butyrate. Moreover, a marked reduction of HNE-induced genotoxicity was caused by preincubation with butyrate. Butyrate also induced the phosphorylation of extracellular signal-regulated kinases (ERK1/2, Western blotting) after 5-30 min, which indicates a regulation of GST expression by this signal pathway. Most effects were greater in HT29 parent cells than in clone cells. In conclusion, butyrate enhances expression of GST and other proteins in both cell lines, which leads to an enhanced chemoprotection, reducing the impact of HNE genotoxicity. Thus butyrate could play a role in early and later stages of cancer prevention by reducing exposure to relevant risk factors.
Ebert
M N
MN
Department of Nutritional Toxicology, Institute for Nutrition, Friedrich-Schiller University, D-07743 Jena, Germany.
Beyer-Sehlmeyer
G
G
Liegibel
U M
UM
Kautenburger
T
T
Becker
T W
TW
Pool-Zobel
B L
BL
D000447:Aldehydes / Q000494:pharmacology*; D016588:Anticarcinogenic Agents / Q000494:pharmacology; D016587:Antimutagenic Agents / Q000494:pharmacology*; D002087:Butyrates / Q000494:pharmacology*; D003106:Colon / Q000187:drug effects* / Q000201:enzymology; D003110:Colonic Neoplasms / Q000235:genetics / Q000517:prevention & control; D004249:DNA Damage / Q000187:drug effects; D004790:Enzyme Induction / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics; D011110:Polymorphism, Genetic; D012333:RNA, Messenger / Q000032:analysis; D014407:Tumor Cells, Cultured
D000447:Aldehydes; D016588:Anticarcinogenic Agents; D016587:Antimutagenic Agents; D002087:Butyrates; D007527:Isoenzymes; D012333:RNA, Messenger; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase; D005978:Glutathione; C027576:4-hydroxy-2-nonenal