Folate deficiency diminishes the occurrence of aberrant crypt foci in the rat colon but does not alter global DNA methylation status.
OBJECTIVE
It has been suggested that a diminished folate status may enhance colorectal carcinogenesis by causing DNA hypomethylation. The aims of the present study were to assess the impact of different levels of folate depletion on azoxymethane (AOM)-induced aberrant crypt foci (ACF) formation and DNA hypomethylation in the colon of male Sprague-Dawley rats.


METHODS
Rats, aged 4 weeks, were divided into four groups and were fed semipurified diets either containing adequate folate (control), devoid of folate (FD) or FD + 1% succinylsulfathiazole before AOM treatment (FD1) or during the last 4 weeks of the study (FD2). At 8 weeks of age, all animals received subcutaneous injections of AOM once weekly for 3 weeks at a dose rate of 15 mg/kg bodyweight. Animals were necropsied 6 weeks after the last AOM injection and the ACF were visualized under light microscopy in formalin-fixed, methylene blue-stained colons.


RESULTS
Blood folate concentrations were significantly depleted (P < 0.001) in the treatment groups consuming folate deplete diets, with the FD2 treatment group having significantly lower folate levels compared with all other groups. Higher plasma homocysteine concentrations (P < 0.001) were observed in the groups that exhibited diminished blood folate levels. There were no significant differences in global DNA methylation in the liver or colonic mucosa between the four groups, despite some groups exhibiting marked folate depletion. Animals with the most severe folate deficiency (FD2) had a lower final bodyweight and had significantly fewer ACF in their colon (P < 0.05) compared with control animals. Total (mean +/- SEM) ACF counts were as follows: control 286+/-24; FD 290+/-25; FD1 218+/-32; and FD2 205+/-27.


CONCLUSIONS
In this model, folate deficiency diminished the occurrence of ACF but did not alter global DNA methylation status in the colon.
Le Leu
R K
RK
University of Adelaide, Department of Medicine, tCSIRO, Health Sciences and Nutrition, South Australia, Australia. richard.leleu@dhn.csiro.au
Young
G P
GP
McIntosh
G H
GH
D000818:Animals; D001397:Azoxymethane / Q000008:administration & dosage; D002273:Carcinogens / Q000008:administration & dosage; D002851:Chromatography, High Pressure Liquid; D003106:Colon / Q000378:metabolism; D003110:Colonic Neoplasms / Q000139:chemically induced / Q000209:etiology* / Q000378:metabolism; D019175:DNA Methylation; D003627:Data Interpretation, Statistical; D004195:Disease Models, Animal; D005492:Folic Acid / Q000097:blood; D005494:Folic Acid Deficiency / Q000150:complications*; D006710:Homocysteine / Q000097:blood; D007279:Injections, Subcutaneous; D007413:Intestinal Mucosa / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D011230:Precancerous Conditions / Q000139:chemically induced / Q000209:etiology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012307:Risk Factors; D013997:Time Factors
D002273:Carcinogens; D006710:Homocysteine; D005492:Folic Acid; D001397:Azoxymethane