Micronuclei and carcinogen DNA adducts as intermediate end points in nutrient intervention trial of precancerous lesions in the oral cavity.
In cancer chemoprevention trials, biomarkers as intermediate end points have gained importance. A variety of biomarkers have been proposed as intermediate end points for upper aerodigestive tract cancers. This study was aimed at studying the frequency of micronucleated cells and carcinogen DNA adducts as indicators of DNA damage and intervention end points in chemoprevention trials. Reverse smokers of chutta (rolled tobacco) from four villages numbering 298 in total were selected. Out of these, 150 were supplemented with four nutrients (vitamin A, riboflavin, zinc and selenium) and 148 controls received placebo, one capsule twice a week for 1 year. Slides of buccal smears were prepared and stained with Fuelgen reaction and counterstained with Fast Green and examined microscopically for the presence of micronucleated cells. Oral cell washings were collected and centrifuged. The DNA adducts were evaluated by the 32P post-labelling assay method. Protein and RNA free DNA (adducted) isolated from the cells was digested with MN/SPD and the DNA adducts isolated by the butanol enrichment procedure. The DNA adducts were identified and quantitated by multidimensional chromatography on PEI-TLC sheets by screen enhanced autoradiography and presented as RAL (relative adduct labelling) values. Both the micronuclei and DNA adducts were significantly elevated in subjects with lesions. At the end of 1 year the frequency of micronuclei decreased significantly (P < 0.001) in the supplemented subjects with or without lesions. The DNA adducts in the supplement group at the end of 1 year also reduced significantly. The adducts decreased by 95% in subjects with all categories of lesions and by 72% in subjects without lesions. No such effects were noted in the placebo group. The two biomarkers investigated in the case study appear to be modifiable by the administration of micronutrient supplements.(ABSTRACT TRUNCATED AT 250 WORDS)
Prasad
M P
MP
National Institute of Nutrition, Indian Council of Medical Research, Hyderabad, India.
Mukundan
M A
MA
Krishnaswamy
K
K
D000328:Adult; D000368:Aged; D002610:Cheek; D018736:DNA Adducts / Q000032:analysis / Q000187:drug effects*; D005260:Female; D006801:Humans; D008297:Male; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects*; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000209:etiology / Q000235:genetics / Q000628:therapy*; D011230:Precancerous Conditions / Q000209:etiology / Q000235:genetics / Q000628:therapy*; D012907:Smoking / Q000009:adverse effects; D014131:Trace Elements / Q000627:therapeutic use*
D018736:DNA Adducts; D014131:Trace Elements