cis-hydroxyproline stimulates the growth of rat mammary carcinoma cells.

With the ever-increasing evidence that the extracellular matrix (ECM) can stimulate tumor growth, it follows that inhibiting the synthesis of tumor-derived stroma may be a potential therapeutic target of cancer progression. The proline analog cis-hydroxyproline (CHP), an inhibitor of collagen deposition, was examined for its effects on the growth of clonal tumor cells that differentially produce type IV collagen and laminin. Two separate clones derived from rat mammary carcinoma cells that produce high and low amounts of type IV collagen and laminin were injected into the flanks of nude mice. Tumors in animals receiving CHP treatment grew faster than tumors in control animals receiving saline, although statistically not significant. Furthermore, upon administration of CHP to these clones in culture, increased proliferation rates of both cell types were observed. These results show that CHP is not useful in preventing stromal development and growth of rat mammary tumor xenografts.

Chronic energy restriction versus energy cycling and mammary tumor promotion.

Chronic energy restriction significantly inhibits mammary tumor promotion in rodents. The present work studied the effect of short-term, intermittent energy restriction or energy cycling on mammary tumor promotion since this feeding paradigm mimics the dieting habits of humans. Female Sprague-Dawley rats were given 7,12-dimethylbenz[a]anthracene (DMBA) at 50 days of age (5 mg ig). One week later, rats were randomly divided into three dietary groups. One group was fed ad libitum throughout the study (AL), another (ER) was fed 40% fewer calories than the AL group, and a third, energy-cycled group (EC) was fed in repeated cycles of 2 days of feeding at a level comparable to that of AL rats followed by 2 days of 40% energy restriction. At 10 weeks post-DMBA, the mammary tumor incidences in the AL and EC groups were the same, but incidence in the ER group was significantly lower. A second experiment examined serum levels of three hormones thought to play a role in mammary tumorigenesis. After 12 or 24 days on diet, ER rats had lower insulin levels compared with the other groups. Serum insulin levels in AL and EC rats were the same. After 24 days on diet, estradiol levels were significantly lower and corticosterone levels higher in the ER and EC groups compared with the AL group. Although energy cycling is a type of energy restriction that lowers overall weight gain and energy intake, it does not inhibit mammary tumor promotion as does chronic energy restriction. These data also suggest that feed efficiency and serum insulin levels correlate with susceptibility to mammary tumor promotion.

Failure of high fat diets to promote mammary cancers in spontaneously hypertensive rats.

Rat strains differ in their susceptibilities to chemically-induced mammary carcinogenesis. The present study tested the hypothesis that the spontaneously hypertensive rat (SHR) strain is resistant to mammary carcinogenesis. Resistance would imply the presence of the mammary carcinoma suppressor (MCS) gene. Therefore, we also wanted to test the ability of this gene to inhibit mammary tumor promotion in the presence of high fat diets. Female SHRs were treated with DMBA (5 mg/rat) at 50 days of age and transferred to either a 20% corn oil or 19% menhaden oil + 1% corn oil diet one week later. At 17 weeks post-DMBA none of the rats in either group developed mammary carcinomas. Multiple palpable nodules formed in the mammary gland indicating that initiation had occurred. We conclude that the SHR strain is genetically resistant to DMBA-induced mammary carcinogenesis by mechanisms involving a blockade of promotion.

High fish oil diet increases oxidative stress potential in mammary gland of spontaneously hypertensive rats.

1. The purpose of this study was to determine whether high omega-3 (19% menhaden oil, 1% corn oil) or high omega-6 (20% corn oil) fatty acid diets would decrease expression of hypertension in the female spontaneously hypertensive rat (SHR), promote tumourigenesis in the rat 7,12-dimethylbenz[a]anthracene (DMBA) model of mammary cancer or increase the susceptibility of the mammary gland to lipid peroxidation. A group of rats on a 5% corn oil diet served as the low fat control group. 2. We found that the high omega-3 and high omega-6 fatty acid diets did not significantly decrease mean arterial pressure. Marked differences occurred between the effects of omega-3 and omega-6 high fatty acid diets on baseline oxidation, auto-oxidation and iron-ascorbate catalyzed oxidation. The omega-3 diet showed 675% increase in basal oxidation, a 2624% increase in auto-oxidation and a 4244% increase in iron-ascorbate catalyzed oxidation compared to the omega-6 diet in mammary tissue homogenates. Although all rats were given 5 mg DMBA (i.g.), no mammary tumours were observed in any of the dietary groups. 3. We conclude that: (i) high polyunsaturated fatty acid diets do not decrease blood pressure in the female SHR; (ii) high fish oil diet markedly increases oxidative potential in the mammary gland; and (iii) the female SHR is resistant to DMBA-induced tumourigenesis.

The effects of patterned calorie-restricted diets on mammary tumor incidence and plasma endothelin levels in DMBA-treated rats.

Chronic caloric restriction has been shown to inhibit mammary tumor promotion in the 7,12-dimethyl-benz[a]anthracene (DMBA) rat mammary tumor model. The objectives of this study were to determine (i) the effects of chronic caloric cycling (yo-yo dieting) on mammary tumor promotion by high fat diets and (ii) the effect of three dietary regimens +/- superimposed mammary tumor burden on plasma endothelin-1,2 (ET) levels. Female Sprague-Dawley rats were treated with DMBA (5 mg/rat) and divided into three dietary groups: ad libitum (AL) (containing 15% corn oil); 40% calorie restricted (CR) (containing 20% corn oil so consumption of fat was equivalent between AL and CR); a calorie cycled (CC) group fed alternatively AL and CR diets each 48 h period. After 10 weeks, tumor incidences were: AL, 63%; CR, 27%; CC, 57% (AL versus CR, P < 0.05; CC versus CR, P < 0.05; AL versus CC, NSD). ET levels (pg/ml plasma) were: AL, 16.0 +/- 6.54; CR, 32.31 +/- 0.34; CC, 23.44 +/- 5.04 (AL versus CR, P < 0.01; CC versus CR, P < 0.01; AL versus CC, P < 0.05). Plasma ET levels were independent of tumor incidence and tumor burden, but plasma ET levels were significantly increased in rats with a prior history of calorie restriction. As expected, maintained caloric restriction reduced mammary tumor incidence but intermittent caloric restriction (caloric cycling or yo-yo dieting) was without similar benefit.

Long-term feeding of corn oil, beef tallow, or menhaden oil and eicosanoid levels in BHE/cdb rats.

The effects of long-term feeding of 2 or 10% fat diets containing corn oil, beef tallow, or menhaden oil on the levels of eicosanoids in brain, plasma, and kidney medulla were studied. Male BHE/cdb rats, which carry a genetic trait for non-insulin-dependent diabetes mellitus, were fed these diets for 9 mo, at which time their glucose tolerance levels were determined, as were brain, kidney medulla, and plasma levels of PGE2, 6-keto-PGF1 alpha, and LTB4. Glucose tolerance was abnormal in the 2 and 10% corn oil groups and normal in the 10% menhaden oil groups. The glucose tolerance levels of the other groups were only slightly disturbed. The levels of LTB4 in kidney and plasma were not affected by dietary fat type or amount. LTB4 levels were significantly higher in the brains of rats fed a low level of beef tallow than in the brains of rats fed a higher level of beef tallow or a low level of menhaden or corn oil. LTB4 values for the brains of rats fed the other diets were intermediate and not different from the aforementioned values. The levels of 6-keto-PGF1 alpha were highest in rats fed the 10% corn oil diet regardless of the tissue assayed. Rats fed the 2 or 10% beef tallow or menhaden oil were not different but had lower levels of this eicosanoid metabolite than rats fed the corn oil diet. PGE2 levels followed the same pattern, suggesting that the impaired glucose tolerance of the corn oil-fed rats had a strong influence on the tissue levels of these eicosanoids.

Eicosanoid synthesis and ornithine decarboxylase activity in mammary tumors of rats fed varying levels and types of N-3 and/or N-6 fatty acids.

Virgin female Sprague-Dawley rats (50 days of age) were administered a single intragastric 10 mg dose of 7,12-dimethylbenz(a) anthracene (DMBA). Three weeks later they were placed on diets containing either 20% corn oil (CO), 20% primrose oil (PO), 20% black currant seed oil (BCO), 20% borage oil (BO), 15% menhaden oil plus 5% corn oil (15% MO + 5% CO), 10% menhaden oil plus 10% corn oil (10% MO + 10% CO), 5% menhaden oil plus 15% corn oil (5% MO + 15% CO) or 10% menhaden oil plus 10% borage oil (10% MO + 10% BO). Incidences of mammary tumors at 16 weeks post-DMBA were 80% in rats fed the CO diet, 84% in rats fed PO diet, 67% in rats fed BCO diet, 88% in rats fed BO diet, 60% in rats fed 15% MO + 5% CO diet, 67% in rats fed 10% MO + 10% CO diet, 83% in rats fed 5% MO + 15% CO diet, and 92% in rats fed 10% MO + 10% BO diet. Tumor multiplicity was lowest in PO-fed rats and highest in BO-fed rats. The tumor burden per tumor-bearing rat was lowest in rats fed the 15% MO + 5% CO, and 10% MO + 10% CO, diets and highest in those fed 20% BCO diet. Although body weight at 16 weeks post DMBA was not significantly different among the dietary groups, food intake was significantly greater in rats fed a diet containing 20% BO, or 5% MO + 15% CO.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of D,L-2-difluoromethylornithine and indomethacin on mammary tumor promotion in rats fed high n-3 and/or n-6 fat diets.

Virgin female Sprague-Dawley rats (50 days of age) were administered a single intragastric 10-mg dose of 7,12-dimethylbenz(a)anthracene (DMBA). Twenty-one days later they were placed on diets containing either 20% corn oil (CO), 15% menhaden oil plus 5% corn oil (MO + CO), 20% CO plus 0.5% w/w of the irreversible ornithine decarboxylase inhibitor, D,L-2-difluoromethylornithine (CO + DFMO), 20% CO plus 0.004% w/w of the cyclooxygenase inhibitor indomethacin (CO + INDO), 20% CO + 0.004% INDO + 0.5% DFMO (CO + INDO + DFMO), or 15% MO + 5% CO + 0.5% DFMO (MO + CO + DFMO). The incidence of DMBA-induced mammary tumors was significantly reduced in rats fed diets containing DFMO but not in rats fed the diet containing indomethacin. Incidences of mammary tumors at 16 weeks post-DMBA were 86% in rats fed the CO diet, 83% in rats ingesting the diet containing CO + INDO, 28% in rats fed CO + DFMO, 32% in rats fed diet containing CO + INDO + DFMO, 59% in rats fed the MO + CO diet, and 24% in rats fed the MO + CO + DFMO diet. The average number of tumors and tumor burden per tumor-bearing rat were reduced and tumor latency was increased in all rats fed diets containing DFMO. Body weight gain, but not food intake, of rats fed the 20% fat + 0.5% DFMO diets was significantly less than in rats fed the 20% fat diets. Prostaglandin E and leukotriene (LTB4) syntheses, ODC activity and mammary tumorigenesis were significantly inhibited by feeding the diet containing menhaden oil or by adding 0.5% DFMO to any of the high fat diets. Feeding a 20% CO diet containing 0.004% INDO significantly reduced prostaglandin synthesis and ODC activity and increased LTB4 synthesis of mammary tumors but did not inhibit mammary tumorigenesis. This study suggests that the 5-lipoxygenase product LTB4 may be involved in mammary tumor production. Whereas a decrease in LTB4 appears to be associated with a decrease in tumorigenesis, an increase (as seen in the indomethacin group) was not associated with any change in the tumorigenic response.

Eicosanoid synthesis in 7,12-dimethylbenz(a)anthracene-induced mammary carcinomas in Sprague-Dawley rats fed primrose oil, menhaden oil or corn oil diet.

The comparative effects of high-fat diets (20%, w/w) on eicosanoid synthesis during mammary tumor promotion in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rats were studied using diets containing 20% primrose oil (PO), 20% menhaden oil (MO) or 20% corn oil (CO). Sprague-Dawley rats fed the PO or MO diet had 21% of 24% fewer adenocarcinomas, respectively, than rats fed the CO diet. Histologically (i.e., mitotic figures, inflammatory cell infiltration and necrosis), the CO-fed rats exhibited the highest frequency of changes within tumors. Plasma fatty acid composition was significantly altered by diet, reflecting the composition of the oils which were being fed. Only the plasma of PO-fed rats contained detectable levels of gamma-linolenic acid (GLA). Arachidonic acid (AA) levels were significantly higher (p less than 0.05) in PO-fed than in CO- or MO-fed rats. MO-fed rats had significantly higher levels of plasma palmitic acid, while palmitoleic, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids were detected only in MO-fed rats. As expected, linoleic acid (LA) and AA levels were lower (p less than 0.05) in the MO-fed rats than in PO- or CO-fed groups. The plasma of the CO-fed rats contained significantly higher levels of oleic acid. Eicosanoid synthesis in mammary carcinomas of rats fed the 20%-fat diets was 2-10 times higher than in mammary fat pads of control rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of 7,12-dimethylbenz(a)anthracene administration on hepatic drug metabolizing enzymes in female rats fed corn, primrose, and menhaden oils.

Female rats receiving a diet containing 20% menhaden oil beginning at 10 weeks of age and continuing for 13 weeks had hepatic benzo(a)pyrene [B(a)P] hydroxylase activity significantly higher than similar rats fed diets containing 20% corn oil or 20% oil of evening primrose. Compared to microsomes recovered from rats fed the corn oil diet, a significant increase in microsomal cytochrome P-450 content along with an increase in the activity of cytochrome P-450 mediated ethoxycoumarin O-dealkylase was evident in rats fed menhaden oil. Glutathione S-transferase activity of the cytosol of hepatocytes was increased twofold by the feeding of 20% menhaden oil, compared with the feeding of corn or primrose oil. Administration of 7,12-dimethylbenz(a)anthracene (DMBA) 21 days before instituting the diets enhanced B(a)P hydroxylase in all animals, with the activity greatest in those fed the menhaden oil. DMBA also caused a significant increase in ethoxycoumarin O-dealkylase in rats fed menhaden oil.

Effects of dietary primrose oil on mammary tumorigenesis induced by 7,12-dimethylbenz(a)anthracene.

The mammary tumor-promoting effect of a high-fat diet containing 20% evening primrose oil (PO) was compared to that of a 20% corn oil (CO) diet. Mammary tumors were induced in female Sprague-Dawley rats using 10 mg (Study 1) and 5 mg (Study 2) 7,12-dimethylbenz(a)anthracene (DMBA). The 10 mg dose of DMBA gave a total mammary tumor incidence of 47% in rats fed the PO diet and 80% for those fed the CO diet. When only adenocarcinomas were counted, the malignant mammary tumor incidences were 41% in rats fed the PO diet and 73% in rats fed the CO diet. In a second study using 5 mg DMBA to induce mammary tumors, total tumor incidences were 50% for PO-fed rats and 63% for those receiving a CO diet. Again, when only adenocarcinomas were counted, tumor incidences were 27% for PO- and 63% for CO-dieted rats. Analysis of plasma fatty acid profiles indicated that animals fed a 20% PO diet showed significant increases in 18:3 and 20:4 fatty acids and significant decreases in 16:0 and 18:1 compared to animals fed a 20% CO diet. These results indicate that the mammary tumor promoting effect of a diet containing 20% fat can be diminished by substituting PO for CO. Moreover, the promoting effect on mammary cancer by a high-fat diet could be depressed by feeding a source of gamma-linolenic acid (GLA).

Effect of thiamin deprivation on the in vitro metabolic activation of benzo(a)pyrene.

Feeding a thiamin-deficient diet to male and female rats for 3 weeks alters the mixed function oxidases responsible for metabolizing benzo(a)pyrene and enhances the response of these enzymes to induction by phenobarbital or 3-methylcholanthrene. The caloric restriction observed in thiamin deprivation may be partially responsible for the enhanced metabolism in this condition but, as established in pair-feeding studies, was not responsible for the enhanced response to enzyme inducers seen in these animals. The degree of altered response was also seen to depend on the sex of the rat and on the substrate concentration of the incubation mixture.

The effects of dietary corn oil on the metabolism and mutagenic activation on N-nitrosodimethylamine (DMN) by hepatic microsomes from male and female rats.

Microsomes from male and female rats fed a diet containing 10% corn oil metabolized N-nitrosodimethylamine (DMN) more rapidly than microsomes from rats fed a similar diet devoid of corn oil. The daily administration of phenobarbital for 4 days prior to harvesting the microsomes resulted in significant induction of the Vmax for N-demethylation of DMN in rats fed the fat-free diet but resulted in no induction (females) or suppression (males) of this enzyme in rats fed the diet containing corn oil. Using concentrations of DMN ranging from 12 to 100 mM, microsomes from rats fed the high fat diet activated DMN to produce mutagenesis in S. typhimurium (TA100) more rapidly than those from rats fed the fat-free diet. Phenobarbital administration induced this activation more effectively in rats fed the corn oil diet than in rats fed the fat-free diet. Phenobarbital induces DMN N-demethylation in rats fed both fat-free and 10% corn oil diets when the DMN concentration is above 10 mM and explains, at least in part, this enhanced mutagenic activation.

Histamine biosynthesis in shock.

The histidine decarboxylase activity of the lung and spleen was determined in rats made resistant to trauma either by prior sublethal exposure or by injection of extracts prepared from the spleens and plasma of trauma-resistant rats. The data describe the posttraumatic period in the normal animal as being associated with an increased histidine decarboxylase activity. In trauma-resistant animals, changes in the enzyme activity were prevented in the lung and were less pronounced in the spleen. The administration of extracts from trauma-resistant rats was similarly effective in impeding the changes in enzyme induction following trauma. It is suggested that an active humoral factor previously shown to be elaborated during conditioning and associated with the RES may act by inhibiting the activation of histidine decarboxylase.