Effects of treatment of rats with indole-3-carbinol on apoptosis in the mammary gland and mammary adenocarcinomas.
Induction of apoptosis is an approach to suppress carcinogenesis. The effects of a 12-week treatment of female Sprague-Dawley rats with indole-3-carbinol (I3C), beta-naphthoflavone or vehicle (40% ethanol in corn oil), by oral gavages starting 3 weeks after initiation of mammary tumorigenesis with 7,12-dimethylbenz[alpha]anthracene, on apoptotic activities in the mammary adenocarcinomas were examined. Apoptotic cells in tumor sections were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and quantitated by light microscopy and an Image-Plus Program. Activities of caspase-3, caspase-8 and caspase-9 were determined by colorimetric assays using the specific substrate and total tumor protein. There were no significant treatment-related effects on the numbers of apoptotic cells and caspase activities in the mammary adenocarcinomas. Likewise, protein expression levels of Bcl-2 and Bax genes in these tumors, determined by Western blot analysis, showed no treatment-related stimulation of apoptotic process. In the absence of tumorigenesis, the activities of caspase-3, caspase-8 and caspase-9 were increased up to approximately 3.6-fold in the mammary gland of rats treated with I3C at 5 or 25 mg/kg of body weight for 4 or 10 days. The I3C-effected induction of caspase-3 activity in the mammary gland was further confirmed by the cleavage of poly (ADP-ribose) polymerase. Treatment of rats with 3,3'-diindolylmethane, a major product of I3C in vivo, at the dose levels equimolar to those of I3C above, did not increase the caspase activities in the mammary gland. Thus, this I3C dimer does not seem to account for the increases of apoptotic activities in the mammary gland observed with I3C. The results suggest that increase of apoptosis in the mammary gland induced by I3C before initiation of tumorigenesis may contribute to suppression of tumor development.
Zhang
Xuexian
X
Veterans Affairs Medical Center (151), Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN, USA. zhang112@umn.edu
Malejka-Giganti
Danuta
D
D015127:9,10-Dimethyl-1,2-benzanthracene; D000230:Adenocarcinoma / Q000188:drug therapy* / Q000473:pathology; D000818:Animals; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002273:Carcinogens; D005260:Female; D007211:Indoles / Q000494:pharmacology*; D008321:Mammary Glands, Animal / Q000166:cytology / Q000187:drug effects; D008325:Mammary Neoplasms, Experimental / Q000139:chemically induced / Q000473:pathology* / Q000517:prevention & control; D051381:Rats; D017207:Rats, Sprague-Dawley; D019324:beta-Naphthoflavone / Q000494:pharmacology
D016588:Anticarcinogenic Agents; D002273:Carcinogens; D007211:Indoles; D015127:9,10-Dimethyl-1,2-benzanthracene; D019324:beta-Naphthoflavone; C016517:indole-3-carbinol