Showing papers by "Bandaru S. Reddy published in 2001"

Modulation of experimental colon tumorigenesis by types and amounts of dietary fatty acids

Abstract: Epidemiological studies and laboratory animal model assays suggest that a high intake of dietary fat promotes colorectal cancer. Several in vivo and in vitro studies support the hypothesis that omega-6 fatty acids promote colon tumorigenesis, whereas omega-3 fatty acids lack promoting activity. Fat intake in the United States traditionally includes high amounts (30% of total caloric intake) of saturated fat rather than omega-6 fatty acids. Therefore, the present study was designed to compare the modulatory effects of a high-fat diet containing mixed lipids (HFML), a diet rich in saturated fatty acids (the average American diet), a diet with fish oil (HFFO) that is rich in omega-3 fatty acids, and a low-fat corn oil diet (LFCO) on the formation of chemically induced colonic aberrant crypt foci (ACF) and tumors, cyclooxygenase (COX)-2 activity, and apoptosis during experimental colon carcinogenesis. At 5 weeks of age, groups of male F344 rats were fed a 5% corn oil diet (LFCO). At 7 weeks of age, rats intended for carcinogen treatment received s.c. injections of azoxymethane at a dose level of 15 mg/kg of body weight once weekly for 2 weeks. Beginning 1 day after the carcinogen treatment, groups of rats were then maintained on experimental diets containing 20% HFML or 20% HFFO. Rats were killed at 8, 23, or 38 weeks after azoxymethane treatment. Colonic ACF and tumors were evaluated histopathologically, and apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated nick end labeling method. Colonic mucosae and tumor samples harvested at week 38 were analyzed for COX-2 synthetic activity and expression. The rats fed the HFML diet showed significantly increased total colonic ACF (P or = 4 aberrant crypts/focus (P or = 4) in multicrypt foci in rats given the HFML diet as compared with such foci in rats fed the HFFO or LFCO diets. By week 23, the HFML diet had significantly increased the incidence of colonic tumors (30-60%) and their multiplicity (100-141%) when compared with the effects of the LFCO or HFFO diets. At week 38, the HFML diet had induced 100% colon tumor incidence and a 4-fold multiplicity of adenocarcinomas compared with the LFCO and HFFO diets. At weeks 23 and 38, a significantly lower percentage of apoptotic colonic epithelial cells were observed in the tumors of animals fed the HFML diet as compared with those fed the HFFO diet. The HFML diet caused significantly increased levels of COX-2 activity in colon tumors (P < 0.05-0.01), and these tumors had enhanced levels of COX-2 expression as compared with those in assays with LFCO or HFFO diets. These observations demonstrate for the first time that HFML diets containing high levels of saturated fatty acids (such as those in Western diets) promote colon carcinogenesis. Although the mechanisms involved in colon tumor promotion by a HFML diet are not fully known, our results indicate that the modulation of eicosanoid production via the influence on COX activity and the suppression of apoptosis may play a key role in HFML diet-induced colon tumorigenesis.

Docosahexaenoic acid regulated genes and transcription factors inducing apoptosis in human colon cancer cells

Abstract: Epidemiological and preclinical studies demonstrate that consumption of diets high in omega-3 fatty acids (n-3 PUFAs) reduce the risk of colon cancer. Docosahexaenoic acid (DHA), a long chain polyunsaturated fatty acid (PUFAs) is a major constituent of nutrients rich in n-3 PUFAs. There are studies to indicate that colon tumor inhibition by n-3 PUFA-rich diets is, in part, mediated through modulation of signaling pathways that alter gene expression which are involved in colon tumor growth. In the present study using CaCo-2 colon cancer cell lines we examined the effects of DHA on the genetic precursors of human colon cancer at the transcription level using DNA oligonucleotide arrays. Our results indicated that DHA inhibits the growth of CaCo-2 cells and induces apoptosis. For gene expression analysis using DNA microarrays, total RNA extracted from DHA treated CaCo-2 cells was converted to cDNA, labeled with Cy5-dCTP (DHA-treated) and Cy3-dCTP (untreated cells) and used as probes for hybridization in human chip spotted with 3,800 oligonucleotides consisting of 156 functional categories. The expression profiles of genes indicated a reprogramming pattern of previously known and unknown genes and transcription factors that provided clues to the possible functional mechanism of DHA. An average of (ratios from triplicate experiments) 504 out of 3,800 genes expressed after 48 h of DHA treatment. Altered expression on the transcription factors includes down regulation of nine members of the RNA II polymerases, transcription co-repressor associated protein and enhancer binding proteins such as AP2, in addition to changes in the expression of zinc finger group of transcription factors. Activation of cytochrome c which triggers caspases was associated with the elevated expression of pro-apoptotic caspases 10, 13, 8, 5 and 9 in DHA treated cells. Activation of cyclin-dependent kinase inhibitors such as p21 (waf1/cip1), p27, p57, p19 and growth arrest specific proteins by more than 2-fold is consistent with the induction of apoptosis and inactivation of antiapototic Bcl-2 family of genes. Inactivation of prostaglandin family of genes, lipoxygenases and altered expression of peroxisome proliferators (PPARalpha and gamma) by DHA seem to indicate a lipid peroxidation-induced apoptosis in addition to effect reflected on the modification of cell cycle regulatory genes. These findings support the conclusion that a genomewide expression profiling of human colon cancer precursor genes and transcription factors provides a set of novel regulatory mechanism(s) to determine the chemopreventive efficacy of DHA and thus to prevent the inflammation and neoplasia.

Chemoprevention of colon cancer by a glutathione conjugate of 1,4-phenylenebis(methylene)selenocyanate, a novel organoselenium compound with low toxicity.

Abstract: We have consistently shown that several synthetic Organoselenium compounds are superior cancer chemopreventive agents and less toxic than selenite or certain naturally occurring selenoamino acids. 1,4-Phenylene bis (methylene)selenocyanate ( p -XSC) is the lead Organoselenium compound in that it has been shown to be the most effective and the least toxic agent in several experimental cancer models. It is not known whether p -XSC or one of its metabolites is responsible for its chemopreventive efficacy. As an initial step, we synthesized one of its putative metabolites, i.e. , the glutathione conjugate of p -XSC ( p -XSe-SG), and determined its stability in the pH range from 2 to 8 and in the diet under normal feeding conditions. We also assessed its maximum tolerated dose and examined its chemopreventive efficacy against azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. p -XSe-SG proved to be very stable over the pH range tested. The maximum tolerated dose of p -XSe-SG determined in a 6-week subchronic toxicity study was found to be >210 ppm (>40 ppm selenium) when the compound was added to AIN-76A high-fat diet. To assess the efficacy of this agent in the postinitiation period of colon carcinogenesis, male F344 rats 6 weeks of age were fed the high-fat diet, and at beginning of weeks 7 and 8, all of the rats intended for carcinogen treatment were given AOM at a dose of 15 mg/kg body weight by s.c. injection. Two days after the carcinogen treatment, the groups of rats consuming the high-fat control diet began their respective high-fat experimental diet regimens with 0, 56, or 84 ppm p -XSe-SG (0.1, 10, and 15 ppm of selenium) supplementation. All animals continued on their respective diets for 38 weeks after the AOM-treatment and were then killed. Colon tumors were evaluated histologically using routine procedures and were also analyzed for cyclooxygenase (COX)-1 and COX-2 expression and enzymatic activities. The results indicate that p -XSeSG administered during the post-initiation stage significantly inhibited both the incidence ( P P P p -XSe-SG in the diet significantly suppressed total COX activity ( P P p -XSe-SG, is stable in the diet and at wide pH ranges, inhibits colon carcinogenesis when administered during the postinitiation stage, and inhibits COX activity. Compared with previous efficacy studies and considering the toxicity associated with selenium, p -XSe-SG seems to be the least toxic Organoselenium chemopreventive agent thus far tested in the experimental colon carcinogenesis. Studies are in progress to delineate whether p -XSe-SG is also effective when administered during the progression stage of colon carcinogenesis.

Multiorgan sensitivity to anticarcinogenesis by the organoselenium 1,4-phenylenebis(methylene)selenocyanate.

Abstract: The data in this report clearly indicate that the form (structure) in which selenium is used is the most critical determinant of success in future clinical trials. Synthetic organoselenium compounds can be tailored to achieve greater chemopreventive efficacy with minimal toxic side effects by structural modifications. We demonstrated that 1,4-phenylenebis(methylene)selenocyanate is a powerful chemopreventive agent against the development of experimental colon, mammary, lung, and oral carcinogenesis. On the basis of metabolism studies of organoselenium compounds and those reported in the literature, our working hypothesis is that aromatic selenol intermediates are important entities in cancer chemoprevention. In addition, we suggest that 1,4-phenylenebis(methylene)selenocyanate not only serves as a chemopreventive agent, but it may be valuable in preventing metastatic diseases in future studies in the clinic.

Inhibitors of inducible nitric oxide synthase for chemoprevention and treatment of cancers

Abstract: Agents and methods for chemoprevention and treatment of neoplasia are described, the agents including a selective inhibitor of inducible nitric oxide synthase and a combination of a selective inhibitor of inducible nitric oxide synthase and an inhibitor of cylcooxygenase-2 in a pharmaceutical composition. The agents and methods are used for chemoprevention and treatment of neoplasia including colorectal cancer and other cancers affecting epithelial cells throughout the body. The agents can also be used to treat the fibrosis that occurs with radiation therapy, as well as adenomatous polyps, including those with familial adenomatous polyposis (FAP).

Modulation of inducible nitric oxide synthase expression in rat intestinal cells by colon tumor promoters.

Abstract: Metabolic epidemiological studies in humans and laboratory animal models demonstrate that consumption of diets high in fat and low in fiber excrete increased levels of 1,2-diacyl-sn-glycerol (DAG) and secondary bile acids such as deoxycholic acid (DA) and lithocholic acid that have been shown to promote colon carcinogenesis. The secondary bile acids and DAG have been shown to activate protein kinase C (PKC) and induce colonic cell proliferation. A large body of evidence indicates that iNOS, an inducible isoform of nitric oxide synthase, is over-expressed in human colon adenomas and in chemically-induced colon tumors of laboratory animals. However, the precise cascade of intracellular events that leads to the iNOS over-expression remains to be fully explored. In this study, we investigated the relationship between induction of iNOS and activation of the PKC pathway by DAG and DC, in an in vitro system using the rat intestinal cell line, RIE-1. As an initial step, we determined whether lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA) modulate iNOS protein expression in RIE-1 cells. Treatment of RIE-1 cells with LPS and PMA for 4 h significantly elevated the iNOS protein expression. The induction of iNOS by the treatment with LPS/PMA was concentration- and time-dependent. Treatment with LPS/DAG or LPS/DC also caused iNOS over-expression in a concentration- and time-dependent fashion suggesting that DAG and DC induce iNOS activity in intestinal cells. Pretreatment with specific PKC inhibitors, bisindolylmaleimide I or Go 6976, inhibited LPS/PMA, LPS/DAG, or LPS/DC-induced iNOS expression and activity. Extracts of the cells treated with LPS/PMA, LPS/DAG or LPS/DC had a high iNOS activity compared to that of control (p<0.04 to p<0.0001). Taken together, our data suggest a possible role of colon tumor promoters, DAG and DC, for iNOS over-expression through activation of the PKC pathway.

Effect of Types and Amount of Dietary Fat During the Initiation Phase of Hepatocarcinogenesis

Abstract: The effects of various levels of corn oil and lard fed during the initiation stage of azoxymethane (AOM)-induced hepatocarcinogenesis were studied in male Fischer 344 rats. The animals were fed diets containing 5%, 13.6%, and 23.5% corn oil or lard two weeks before, during, and until one week after injections of AOM (15 mg/kg body wt s.c.) once weekly for two weeks. One week after AOM treatment, groups of animals fed the 13.6% and 23.5% corn oil or lard diet were transferred to their respective 5% corn oil or lard diet and fed these diets until the termination of the study (34 wk). Immunohistochemical staining of glutathione S-transferase placental form was performed in the liver, and the number of glutathione S-transferase placental form-positive foci was determined. Density, average area, and unit area of foci were significantly inhibited in the animals fed the 13.6% and 23.5% lard diets compared with those fed the 13.6% and 23.5% corn oil diets. These results indicate that the effect of dietary fat during the initiation phase of AOM-induced hepatocarcinogenesis depends on the type of fat and its fatty acid composition. Additionally, the enhancing effect of a corn oil diet in hepatocarcinogenesis is mainly present during the initiation phase of carcinogenesis compared with a lard diet.