Inhibition by chlorophyllin of 2-amino-3-methylimidazo-[4,5-f]quinoline-induced tumorigenesis in the male F344 rat.

This report describes a 1-year carcinogenicity bioassay in which male F344 rats received 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) by gavage 2-3 times per week throughout the first 35 weeks of study, while 0.1% sodium/copper chlorophyllin (CHL) was given concomitantly in the drinking water. The carcinogen dose was calculated to provide an average exposure of 4.2 mg IQ/rat per day, equivalent to that reported in previous carcinogenicity study in which rats were given 0.03% IQ in the diet. Adjusted for length of follow-up, inhibition was highly significant for total tumor burden (P < 0.001), and for specific sites was significant in the small intestine and Zymbal's gland (P < or = 0.1), and in the liver (P < 0.01). In rats given CHL, the first onset of tumors in the Zymbal's gland was delayed from week 20 to 36, but was shortened from week 31 to 26 in the skin (P < 0.05). These results support a chemopreventive role for CHL in several of the major target organs for IQ tumorigenesis in the rat, but raise concerns over possible deleterious effects in the skin.

Colonic cell proliferation, apoptosis and aberrant crypt foci development in rats given 2-amino-3-methylimidaz.

Sodium-copper chlorophyllin (CHL) inhibits the formation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)- and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced colonic aberrant crypt foci (ACF) and tumours in the F344 rat when it is given simultaneously with either carcinogen. However, CHL reportedly increased the incidence of dimethylhydrazine (DMH)-induced colon tumours in the same species when administered post-initiation. In the present study, rats were given IQ (130 mg/kg body weight, by oral gavages on alternating days) for 2 weeks, starting in experiment week 3, and one week after the final IQ dose rats received CHL treatment until the study was terminated at 16 weeks. Compared with animals given carcinogen alone, the mean number of IQ-induced ACF per colon was reduced significantly by 1% (w/v) CHL in the drinking water (P < 0.05), whereas 0.1% and 0.01% CHL had no effect. These CHL concentrations increased in a dose-related manner both the terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) and bromodeoxyuridine (BrdU) labelling indices in the distal colon. However, the lowest concentration tested, 0.001% CHL, increased the mean number of IQ-induced ACF per colon (P < 0.05), and increased the BrdU labelling index without a concomitant change in TUNEL. These studies indicated that 0.001% CHL promoted IQ-ACF due to deregulation of the homeostatic balance between cell birth and apoptosis in the colonic mucosa, whereas higher concentrations of CHL had either no effect or protected against IQ-induced ACF by causing dose-related increases in the overall rate of cell turnover in the colon.

Post-initiation effects of chlorophyllin and indole-3-carbinol in rats given 1,2-dimethylhydrazine or 2-amino-3-methyl- imidazo.

Chlorophyllin (CHL) is a water-soluble derivative of chlorophyll, the ubiquitous pigment in green and leafy vegetables, whereas indole-3-carbinol (I3C) is present in cruciferous vegetables such as cabbage, broccoli and cauliflower. In rats initiated with 1,2-dimethylhydrazine (DMH), CHL and I3C reportedly promoted or enhanced the incidence of colon tumors when they were administered after, or during and after the carcinogen exposure, respectively. The same compounds given post-initiation inhibited the formation of colonic aberrant crypts induced by heterocyclic amines, such as 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), but tumor suppression was not examined in the latter studies. In the present investigation, male F344 rats were treated with IQ or DMH during the first 5 weeks of a 1 year study; IQ was given in the diet (0.03%), whereas DMH was administered once a week by s.c. injection (20 mg/kg body wt). Beginning 1 week after the last dose of IQ or DMH until sacrifice, rats received 0.001, 0.01 or 0.1% (w/v) CHL in the drinking water or 0.001, 0.01 or 0.1% I3C in the diet. Compared with controls given carcinogen alone, 0.1% I3C treatment suppressed the multiplicity of IQ-induced colon tumors, and CHL inhibited in a dose-related manner the incidence of IQ-induced liver tumors. However, 0.001% CHL increased significantly the multiplicity of DMH-induced colon tumors while having no effect on the colon tumors induced by IQ. These results indicate that both the choice of carcinogen as well as the dose of the tumor modulator can be important determinants of the events that occur during post-initiation exposure to CHL or I3C. Based on the present findings and data in the literature, it is possible for CHL and I3C to act as tumor promoters or anticarcinogens, depending upon the test species, initiating agent and exposure protocol.

beta-Catenin mutation in rat colon tumors initiated by 1,2-dimethylhydrazine and 2-amino-3-methylimidazo[4,5-f]quinoline, and the effect of post-initiation treatment with chlorophyllin and indole-3-carbinol.

Carcinogens 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 1,2-dimethylhydrazine (DMH) induce colon tumors in the rat that contain mutations in beta-catenin, but the pattern of mutation differs from that found in human colon cancers. In both species, mutations affect the glycogen synthase kinase-3beta consensus region of beta-catenin, but whereas they directly substitute critical Ser/Thr phosphorylation sites in human colon cancers, the majority of mutations cluster around Ser33 in the rat tumors. Two dietary phytochemicals, chlorophyllin and indole-3-carbinol, given post-initiation, shifted the pattern of beta-catenin mutations in rat colon tumors induced by IQ and DMH. Specifically, 17/39 (44%) of the beta-catenin mutations in groups given carcinogen plus modulator were in codons 37, 41 and 45, and substituted critical Ser/Thr residues directly, as seen in human colon cancers. None of the tumors from groups given carcinogen alone had mutations in these codons. Interestingly, many of the mutations that substituted critical Ser/Thr residues in beta-catenin were from a single group given DMH and 0.001% chlorophyllin, in which a statistically significant increase in colon tumor multiplicity was observed compared with the group given DMH only. These tumors had marked over-expression of cyclin D1, c-myc and c-jun mRNA and c-Myc and c-Jun proteins were strongly elevated compared with tumors containing wild-type beta-catenin. The results indicate that the pattern of beta-catenin mutations in rat colon tumors can be influenced by exposure to dietary phytochemicals administered post-initiation, and that the mechanism might involve the altered expression of beta-catenin/Tcf/Lef target genes.