Showing papers by "Bandaru S. Reddy published in 1996"

Evaluation of Cyclooxygenase-2 Inhibitor for Potential Chemopreventive Properties in Colon Carcinogenesis

Abstract: Epidemiological and laboratory studies indicate an inverse relationship between the risk of colon cancer development and intake of nonsteroidal antiinflammatory agents, including aspirin. One of the mechanisms by which nonsteroidal antiinflammatory agents inhibit colon carcinogenesis is through the inhibition of prostaglandin production by cyclooxygenase isozymes (COX-1 and COX-2). Overexpression of COX-2 has been observed in colon tumors. Thus, selective inhibitors of COX-2 could potentially serve as chemopreventive agents. We have assessed the chemopreventive properties of SC-58635, a COX-2 inhibitor, and of sulindac, as a positive control, in a double-blind study, using azoxymethane-induced colonic aberrant crypt foci (ACF) as a measure of efficacy. Five-week-old male F344 rats were fed the control diet (modified AIN-76A) or experimental diets containing 150 or 1500 ppm SC-58635, 320 ppm sulindac, or 1500 ppm placebo. Two weeks later, all animals except those in vehicle (normal saline)-treated groups were s.c. injected with azoxymethane (15 mg/kg of body weight, once weekly for 2 weeks). At 16 weeks of age, all rats were sacrificed and colons were evaluated for ACF. As expected, dietary administration of sulindac suppressed ACF development as such and reduced crypt multiplicity in terms of number of aberrant crypts/ focus. Administration of 1500 ppm SC-58635 inhibited total ACF induction and crypt multiplicity by about 40-49%. Our finding that SC-58635 significantly suppressed colonic ACF formation and crypt multiplicity strengthens the hypothesis that a selective COX-2 inhibitor possesses chemopreventive activity against colon carcinogenesis.

Modulating effect of amount and types of dietary fat on colonic mucosal phospholipase A2, phosphatidylinositol-specific phospholipase C activities, and cyclooxygenase metabolite formation during different stages of colon tumor promotion in male F344 rats

Abstract: Epidemiological and laboratory animal model studies suggest that the effect of dietary fat in colon carcinogenesis depends not only on the amount but on its fatty acid composition. Animal model studies demonstrated that high dietary corn oil or safflower oil rich in omega-6 fatty acids increased the colon tumor promotion, whereas diets containing fish oil high in omega-3 fatty acids had no such enhancing effect. One of the mechanisms by which high dietary fat enhances colon carcinogenesis may be through the modulation of colonic mucosal phospholipase A2 (PLA2) and phosphatidylinositol-specific phospholipase C (PI-PLC), which are dominant pathways for arachidonic acid release and formation of eicosanoids. PI-PLC is also responsible for diacylglycerol formation and protein kinase C-dependent signal transduction and cell proliferation. In the present study, we investigated the modulating effect of high fat diets rich in omega-3 and omega-6 fatty acids on colonic mucosal PLA2, PI-PLC activities, and eicosanoid (prostaglandins and thromboxane B2) formation from arachidonic acid via cyclooxygenase (COX) during different stages of azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. At 5 weeks of age, groups of animals were fed the low-fat diet containing 5% corn oil. Beginning at 7 weeks of age, all animals except those intended for vehicle treatment received AOM s.c. once weekly for 2 weeks at a dose rate of 15 mg/kg body weight. Vehicle-treated groups received an equal volume of normal saline. One day after the second AOM or vehicle treatment, groups of animals were transferred to experimental diets containing 23.5% corn oil and 20.5% fish oil + 3% corn oil, whereas one group continued on the low-fat diet containing 5% corn oil. Groups of animals were then sacrificed at weeks 1, 12, and 36 after the second AOM-or saline-treatment. Colonic mucosa harvested at weeks 1, 12, and 36 and colonic tumors obtained at week 36 were analyzed for PLA2, PI-PLC, and eicosanoid formation from arachidonic acid by the action of COX. The results demonstrate that colon carcinogen treatment increases the activities of colonic mucosal PLA2 and PI-PLC and the formation of prostaglandins and thromboxane A2 from arachidonic acid through COX throughout the study period compared to saline-treated animals fed similar diets. The activities of PLA2, PI-PLC, and COX were significantly higher in colon tumors compared to colonic mucosa. These results also demonstrate that a high-fat diet containing corn oil increases colonic mucosal and tumor PLA2 and PI-PLC and the formation of prostaglandins and thromboxane B2 by the action of COX as compared to low dietary corn oil or a diet high in fish oil. The results of our study offer one of the mechanisms by which the amount and types of dietary fat modulate colon carcinogenesis.

Effect of amount and types of dietary fat on intestinal bacterial 7 alpha-dehydroxylase and phosphatidylinositol-specific phospholipase C and colonic mucosal diacylglycerol kinase and PKC activities during stages of colon tumor promotion.

Abstract: It is evident from many studies that the effect of dietary fat on colon tumor promotion depends not only on the amount of fat but especially on fatty acid composition. Animal model studies have shown that diets which are high in omega-6 fatty acids increase colon tumor promotion, whereas diets rich in omega-3 fatty acids have no such enhancing effect. The mechanisms by which the high fat content of the diet promotes colon carcinogenesis may include the production of secondary bile acids in the colon and the modulation of colonic luminal bacterial 7 alpha-dehydroxylase that is involved in generating secondary bile acids, phosphatidylinositol-specific phospholipase C (PI-PLC), and mucosal PI-PLC, as well as diacylglycerol (DAG) kinase and protein kinase C (PKC). In the present study, we investigated the effect of high-fat diets that are rich in omega-3 and omega-6 fatty acids on cecal bacterial 7 alpha-dehydroxylase and PI-PLC, fecal secondary bile acids, and colonic mucosal DAG kinase and PKC activities during different stages of colon carcinogenesis in male F344 rats. At 5 weeks of age, groups of animals were fed a low-fat diet containing 5% corn oil (LFCO). Beginning at 7 weeks of age, all animals, except those intended as vehicle controls, received azoxymethane (AOM) s.c. once weekly for 2 weeks at a dose rate of 15 mg/kg body weight. Vehicle-treated groups received s.c. injections of normal saline. One day after the second AOM or saline treatment, the experimental groups of animals were transferred to a high-fat diet containing 23.5% corn oil (HFCO) or 20.5% fish oil + 3% corn oil (HFFO). One group continued on the LFCO diet. Animals were sacrificed at weeks 1, 12, and 36 after the AOM or saline treatment. Colonic mucosa were harvested at weeks 1, 12, or 36, and the colonic tumor tissues were examined for PKC and DAG kinase activities. Contents of the cecum were analyzed for bacterial 7 alpha-dehydroxylase and PI-PLC activities. Stool samples collected at week 12 were analyzed for bile acids. High corn oil content of the diet significantly increased the cecal bacterial 7 alpha-dehydroxylase and PI-PLC activities as compared to the diets with high fish oil or low corn oil content. Animals fed the HFCO diet excreted higher levels of secondary bile acids, such as deoxycholic acid and lithocholic acid, than those fed the LFCO or HFFO diets. Carcinogen treatment significantly enhanced the activities of DAG kinase and total membrane PKC activities in colonic mucosa compared to saline treatment in all dietary groups. Animals treated with saline or AOM and fed HFCO showed increased levels of DAG kinase and membrane PKC activities in the colonic mucosa when compared to LFCO and HFFO groups. DAG kinase and membrane PKC activities were higher in colon tumors than in the surrounding colonic mucosa, and also increased levels of these enzyme activities were found in the HFCO diet group. These results indicate that the modifying effect of dietary fat on colonic bacterial enzymes, secondary bile acids, colonic mucosal and tumor DAG kinase, and PKC that may play a role in colon carcinogenesis depends on the types and amount of fat given. The colon tumor-enhancing effect of a HFCO diet in contrast to the high dietary fish oil may be, in part, explained on the basis of its modulating effect on these bacterial and colonic mucosal enzymes and colonic secondary bile acids relevant to colon tumor promotion.

Regional distribution of carcinogen-induced colonic neoplasia in the rat.

Abstract: Carcinogen induction of neoplasms in rodent colon has been used as a model for human colon cancer development and for evaluating chemopreventive regimens. We studied the regional distribution of small and large intestinal tumors in 229 rats given azoxymethane (AOM) once weekly for two weeks (15 mg/kg sc). The AOM regimen induced 63% more tumors in distal (DC) than in proximal colon (PC), although tumor volume was greater in PC. A high‐fat (23% com oil) diet increased tumors in PC and DC (p < 0.01). Caloric restriction of 10–30% of the ad libitum diet progressively reduced DC tumor formation but did not alter PC tumors. Tumor volume was unaffected by either regimen. Small intestinal tumors were concentrated in the proximal 15 cm of the intestine and were unaffected by dietary manipulation. This AOM model of colon tumor formation approximates human colon cancer distribution and is an appropriate model for rodent chemopreventive studies.

Inhibition of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced lymphoma formation by oltipraz

Abstract: 2-Amino-1-methyl-6-phenylimidazo[4,5- b ]pyridine (PhIP) is a mutagenic and carcinogenic heterocyclic amine present in pyrolysate products of meat and fish and has been shown to induce tumors in the colon, mammary gland, and possibly lymphatic system. Experiments were designed to examine the lymphoma-inducing capacity of PhIP and to test the inhibitory effects of oltipraz on PhIP-induced lymphomas in male F344 rats. Beginning at 5 weeks of age, groups of rats were fed the diets containing 0, 200, and 400 ppm oltipraz with or without 100–400 ppm PhIP. All animals were continued on this regimen until the 58th week. The results indicate that administration of PhIP produced lymphomas in 75% of rats Most of the large lymphomas were thymomas (65%), and these lymphomas developed in less than 6 months. Death of animals during the course of the study was due to suffocation produced by a large lymphoma that filled the entire thoracic cavity, resulting in collapse of the lungs. Administration of 200–400 ppm oltipraz significantly protected rats from PhIP-induced toxicity; most of the rats survived until termination of the experiments. It is noteworthy that the addition of oltipraz at 200 and 400 ppm in the diet suppressed the PhIP-induced lymphomas to 90–100%. In conclusion, PhIP-induced lymphomas in the laboratory rat appears to be a very useful model to analyze the genesis of lymphomas, and oltipraz serves as a potential chemopreventive agent for lymphomas.

Lack of Tumorigenicity of Cholesterol Epoxides and Estrone-3,4-quinone in the Rat Mammary Gland

Abstract: The purpose of this study is to test the long-standing hypothesis that endogenous agents found in human breast fluid and in plasma are potential initiators of breast cancer. Therefore, we evaluated the tumorigenicity in the mammary glands of female CD rats of cholestan-5α,6α-epoxy-3β-ol (cholesterol-α-epoxide), cholestan-5β,6β-epoxy-3β-ol (cholesterol-β-epoxide), and 1,5(10)estradiene-3,14,17-trione (estrone-3,4-quinone). As a positive control, trans -3,4-dihydroxy- anti -1,2-epoxy-1,2,3,4-tetrahydrobenzo[ c ]phenanthrene (BcPDE) was used. Rats were fed a high-fat AIN-76A diet (23.5% corn oil) to mimic the Western dietary composition. Because literature data suggest that the endogenous agents tested in this study are weak electrophiles, the total doses of cholesterol epoxides (12.3 µmol/rat) and of estrone-3,4-quinone (30 µmol/rat) were 10- and 25-fold higher, respectively, than that of BcPDE (1.2 µmol/rat). Each agent was dissolved in DMSO, and one-sixth of the total dose was injected under each of six nipples on the left side of the rat, whereas DMSO only was injected under the nipples on the right side. The thoracic glands of the rat were treated at 30 days of age, and those located in the inguinal area were treated on the following day. The experiment was terminated 44 weeks after treatment. Consistent with our previous study, BcPDE was a strong mammary carcinogen. However, there were no differences between rats treated with DMSO alone and those receiving DMSO containing cholesterol-α-epoxide, cholesterol-β-epoxide, or estrone-3,4-quinone. The results of this study clearly indicate, for the first time, that metabolites derived from cholesterol and estrone lack tumorigenic activity in the rat mammary gland, at least under the conditions of the present protocol.

Effects of dietary fat content on the metabolism of NNK and on DNA methylation induced by NNK

Abstract: The available data support the concept that high-fat diets increase cytochrome P-450 activities in the liver, leading to increased rates of carcinogen metabolism and, in some instances, DNA adduct formation. Therefore we investigated whether a high-fat diet can also influence DNA methylation by the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the lungs of rats. Male F344 rats were fed a regular AIN-76A low-fat (5% corn oil) or AIN-76A high-fat (23.5% corn oil) diet. After three weeks on this dietary regimen, the animals were injected subcutaneously once daily for four days with NNK at 0.39 mmol/kg body wt. Groups of rats were sacrificed 4 and 24 hours after the last NNK administration; livers and lungs were excised for DNA isolation. We found that the high-fat diet significantly enhanced the formation of O6-methylguanine (O6-mGua) in the rat lung four hours (p < 0.01) after the last carcinogen administration. This may, in part, account for our previous finding in regard to the enhancing effect of the high-fat diet on NNK-induced lung carcinogenesis. There was no effect on O6-mGua or 7-mGua in the rat liver at either time point. To further elucidate the enhancing effect of the high-fat diet on DNA methylation by NNK in the lung, we determined its effect on the in vitro and in vivo metabolism of NNK. The in vitro data indicated that dietary fat has no measurable effect on liver and lung microsomal mixed-function oxidase in catalyzing the metabolic activation of NNK. The results of the metabolism study of NNK in vivo appear to be consistent with the in vitro finding, in that fat had no effect on the excretion pattern of NNK or on the distribution pattern of its urinary metabolites. It is apparent that the enhancing effect of the high-fat diet on O6-mGua in the lung of rats that was measured four hours after NNK injection requires future investigations.

Calcium carbonate treatment of diarrhoea in intestinal bypass patients

Abstract: Objective : To study the effect of supplemental calcium carbonate on faecal water, calcium, bile acid and lipid concentration and output in intestinal bypass subjects with diarrhoea. Design : Prospective, single-arm treatment trial of oral calcium carbonate, 2400 or 3600 mg Ca 2+ per day, given for 12 weeks. Methods : Faecal constituents were determined in wet or lyophilized stool of 24-h collections at baseline and at 12 weeks of study. Calcium was measured by absorption spectrophotometry. Bile acids, long-chain fatty acids and short-chain fatty acids were analysed by gas chromatography. Results : In 15 subjects, calcium supplementation reduced bowel frequency by a mean of 49%, faecal wet weight by a mean of 50% (1292 to 646g per day), and dry weight by a mean of 36%, P<0.001. Faecal water concentration was reduced from 76.4 ± 1% to 70.2 ± 2%, P<0.01. Faecal water concentration was inversely correlated with dry faecal calcium concentration (r = -0.75, P=0.00001). Conclusion : Calcium reduces the diarrhoea of intestinal bypass patients. It is suggested that the constipating effect of calcium may be related to reduction in faecal water.