Influence of prolactin and growth hormone on rat mammary tumors induced by N-nitrosomethylurea

The effects of hypophysectomy and prolactin-suppressing drugs on the growth of mammary tumors induced in Sprague-Dawley rats by N-nitrosomethylurea and dimethylbenz(a)anthracene were compared. The influence of ovine prolactin and growth hormone administration on N-nitrosomethylurea-induced tumors was also studied in hypophysectomized animals. After hypophysectomy, all 13 tumors induced in 13 rats by N-nitrosomethylurea underwent regression, as did ten of 12 induced by dimethylbenz(a)anthracene. There were no new tumors. Pergolide mesylate, a long-acting ergoline derivative, was given in a dose of 80 micrograms twice daily by s.c. injection for 28 days. Only three of 12 N-nitrosomethylurea-induced tumors regressed, while four became static. However, only two new tumors developed in the 12 pergolide-treated rats, compared to 11 in the 12 untreated controls. Bromocriptine mesylate, at ten times the pergolide dose, was even less effective; one of 16 tumors regressed, two became static, and eight new tumors appeared in the 16 rats. In contrast, eight of 12 dimethylbenz(a)anthracene-induced tumors regressed during pergolide therapy, two became static, and there was only one new tumor among the 12 rats. Prolactin, 1 mg twice daily for 7 days by s.c. injection, was given to another eight rats bearing 11 N-nitrosomethylurea-induced tumors, commencing 7 days after hypophysectomy. Regression of five tumors borne by four rats was reversed but resumed when treatment was stopped. Regression of five tumors in the other four animals was arrested without regrowth; the sixth became inpalpable. All of these six grew rapidly when growth hormone, 2 mg twice daily, was administered in addition to prolactin.     

Net energy effects of dietary fat on chemically induced mammary carcinogenesis in F344 rats

The effect of net energy, as distinct from kilocalorie intake or the percent of fat in the diet, on 7,12-dimethylbenz[a]anthracene [(DMBA) CAS: 57-97-6]-induced mammary tumorigenesis in female inbred F344 rats was investigated. Rats were fed a 5% corn oil diet from weaning until DMBA administration, when they were switched to one of three dietary regimens: 5% corn oil diet, low-fat diet fed ad libitum (LF); 30% corn oil diet, high-fat diet fed ad libitum (HF); or 30% corn oil diet fed at a level providing a calculated net energy equivalent to the group on LF [high-fat diet fed at a restricted level (HF-R)]. Calculated relative net energy values of the amounts of diet actually consumed by the groups on HF-R, LF, and HF were, respectively, 0.90, 1.00, and 1.07 (kcal equivalent to 34.1, 42.2, and 40.8, respectively). Weight gain for the groups on LF and HF-R was the same throughout the experiment (24 wk), while rats on HF weighed significantly more at 6 weeks and thereafter. Body composition analyses at 24 weeks established that the groups on HF and HF-R were equivalent in fat: protein ratio, whereas the group on LF had about 35% less body fat and 15% more body protein. Carcass energy was in the following order for rats in these diet groups: HF greater than HF-R greater than LF. At 24 weeks, tumor incidences for the groups on HF, LF, and HF-R were, respectively, 73, 43, and 7%. These data indicated that tumor appearance does not depend on the percent fat in the diet per se but rather on a complex interaction involving energy intake, energy retention, and body size.     

Effect of dietary fat on X-ray-induced mammary cancer in Sprague-Dawley rats

We studied the effect of dietary fat levels on the induction of mammary cancer by 350 rads total-body X-radiation given to noninbred albino Sprague-Dawley rats at 50 days of age. Compared to rats on a low-fat (LF) diet (5% lard), rats on a high-fat (HF) diet (20% lard) from 30 days of age had more tumors, with a higher multiplicity of carcinomas per rat. LF-fed groups exhibited a longer median tumor latency period thatn did HF-fed groups. A similar trend toward more tumors with an earlier time of death was seen in rats given single iv doses of 50 mg 1-methyl-1-nitrosourea/kg and fed an HF diet as compared to an LF diet.     

The induction of bladder tumors in F344 rats by intravesicular administration of some nitrosamines

Three nitrosamines, metabolically related and formed in vivo from the bladder carcinogen nitrosomethyl-n-octylamine, were administered to groups of 12 female F344 rats by intraurethral instillation twice a week for 30 weeks. All three compounds induced tumors in the urinary bladder. Nitrosomethyl-2-oxopropylamine at 10 mg/ml was the most potent, causing death of half of the animals with tumors at 43 weeks, following a total dose of 1.0 mmol; most of the rats also had tumors of the nasal mucosa, and there were some tumors of the kidney and kidney pelvis. Nitrosomethyl-2-hydroxypropylamine at 10 mg/ml (total dose 1.0 mmol) was much less effective, the median week of death being 83 weeks. In addition to bladder tumors, this group had tumors of the nasal mucosa, esophagus, and kidney. Nitrosomethyl-3-carboxypropylamine at 75 mg/ml and a total dose of 6.2 mmol per rat induced a high incidence of bladder tumors and tumors of the kidney pelvis, but not tumors of the nasal mucosa; the median week of death for this group was 55 weeks. It is concluded that nitrosomethyl-n-alkylamines that induce bladder tumors by oral administration to rats are metabolized to nitrosomethyl-3-carboxypropylamine, which is excreted in the urine and further metabolized to nitrosomethyl-2-oxopropylamine, the proximate bladder carcinogen.     

Adrenal regulation of mammary tumorigenesis in female Sprague-Dawley rats: histopathology of mammary tumors

Mammary tumors induced in female Sprague-Dawley rats by feeding 7,12-dimethylbenz(a)anthracene (DMBA; 20 mg/100 g body weight) were classified according to histological criteria of tissue differentiation, cellular atypia, and evidence of invasion. The 549 tumors could be placed in three categories, nodular hyperplasia, nodular hyperplasia with atypia, and carcinoma, and combinations of all three. Although tumors classified histologically as carcinomas did not metastasize, upon transplantation to the kidney capsule, a tumor classified as a carcinoma grew for eight generations and metastasized. Tumor heterogeneity was a common finding in DMBA-initiated tumors. Carcinomas were an early lesion. As the length of time between DMBA treatment and sacrifice increased, more tumors with areas of carcinoma were found. Therefore, DMBA-initiated tumors progressed to carcinomas either soon after initiation or later by development within nodular hyperplasias. In 4 separate groups of animals (74 adrenalectomized rats and 90 intact rats), postinitiation adrenalectomy increased the numbers of carcinomas compared to intact animals. This effect was consistently seen in the cervical and thoracic mammary glands. We propose that the mechanism for enhancement of progression to greater malignancy by adrenalectomy may be inhibition of differentiation of initiated cells in the absence of glucocorticoids.     

Interaction of dietary fat and the thymus in the induction of mammary tumors by 7,12-dimethylbenz(a)anthracene

The interaction of dietary fat and the thymus in the induction of mammary tumors by dimethylbenz(a)anthracene has been examined in female Sprague-Dawley rats. In these experiments, rats fed diets of 0.5% (low fat), 5% (normal fat), or 20% (high fat) corn oil from weaning (21 days of age) were thymectomized or sham thymectomized at 35 days of age and were given 5 mg of dimethylbenz(a)anthracene at 55 days of age. Thymectomy exerted a protective effect in rats fed low and normal fat diets, and this was not reversed by Thymosin Fraction V. In high fat-fed rats, tumorigenesis was increased compared to the low fat groups, and in addition, the protective effect of thymectomy was absent. This differential effect of thymectomy could not be explained on the basis of changes in prolactin concentration, since prolactin levels were decreased in all dietary groups. Neither diet nor thymectomy affected corticosterone levels or the estrus cycle of mature rats. Peripheral blood lymphocytes were, however, decreased by both thymectomy and increasing the fat content of the diet. It is hypothesized that the promoting effect of dietary fat on dimethylbenz(a)anthracene-induced mammary tumorigenesis is mediated via the immune system, although a role for the endocrine system still cannot be ruled out.     

Effects of high dietary fat on the growth and development of ovarian-independent carcinogen-induced mammary tumors in rats

This study examined the influence of high dietary fat intake on the development of ovarian-independent mammary tumors in both vehicle-treated controls and rats made deficient in estrogen and prolactin during tumor induction. The majority of 7,12-dimethylbenz(a) anthracene (DMBA)-induced mammary tumors in rats are dependent on estrogen and prolactin for growth, and suppression of prolactin and estrogen at the time of tumor initiation causes a reduction in tumor incidence and increase in tumor latency. However, the majority of mammary tumors which do develop in these animals exhibit ovarian-independent growth. Sprague-Dawley rats were given 7.5 mg DMBA p.o. at 57 days of age. Starting 1 day prior to and continuing for 7 days after DMBA administration, rats were given daily injection of vehicle or the combination of tamoxifen (20 micrograms/rat) plus bromocryptine (5 mg/kg). At the end of drug treatment, rats in each treatment group were equally divided and placed on normal fat (5% corn oil) or high fat (20% corn oil) diets for the duration of the experiment. Vehicle-treated rats were ovariectomized 27 wk and drug-treated rats 47 wk after DMBA administration to determine tumor ovarian dependency. Vehicle-treated rats fed high fat diets showed significant increases in mammary tumor incidence and number as compared to similarly treated rats fed a normal fat diet, with approximately 80% of the tumors in each group being ovarian dependent. Likewise, tamoxifen-bromocryptine-treated rats fed a high fat diet showed a significant enhancement in mammary tumor number, although not incidence, as compared to similarly treated rats fed a normal diet. Tumors in these drug-treated groups displayed essentially the same incidence of ovarian dependence (23%). Tamoxifen-bromocryptine-treated groups displayed a 2-fold increase in latency of tumor appearance as compared to vehicle-treated controls; however, this long latency was not reduced when these rats were fed a high fat diet. These results demonstrate that high dietary fat stimulates ovarian-dependent and -independent mammary tumorigenesis in rats but does not influence the hormonal responsiveness of these tumors.     

Induction of liver tumors in F344 rats by crotonaldehyde

The tumorigenic activities in F344 rats of crotonaldehyde, a representative alpha, beta-unsaturated carbonyl compound, and N-nitrosopyrrolidine, which could produce crotonaldehyde upon metabolism, were compared. Groups of rats were treated with either crotonaldehyde (0.6 mM or 6.0 mM) or N-nitrosopyrrolidine (0.6 mM) in their drinking water for 113 or 84 weeks, respectively. At the 0.6 mM dose, crotonaldehyde induced neoplastic lesions of the liver in 9 of 27 rats; 2 rats had hepatocellular carcinomas, and 9 rats had neoplastic nodules. It also caused altered liver cell foci in 23 of 27 rats. The incidences of tumors and foci were significantly higher than those of the control group. N-Nitrosopyrrolidine induced hepatocellular carcinomas in 20 of 23 rats, liver neoplastic nodules in 16 of 23 rats, and altered liver cell foci in 23 of 23 rats. Thus, crotonaldehyde appears to be a weaker tumorigen than N-nitrosopyrrolidine. At the 6.0 mM dose, crotonaldehyde treatment caused moderate to severe liver damage in 10 of 23 rats. No preneoplastic or neoplastic lesions were observed in these rats. The remaining 13 rats of this group developed altered liver cell foci. The tumorigenicity of crotonaldehyde suggests that alpha, beta-unsaturated carbonyl compounds, which are ubiquitous in the human environment and can be formed endogenously, may be an important class of potential carcinogens.     

Effects of arctiin on PhIP-induced mammary, colon and pancreatic carcinogenesis in female Sprague-Dawley rats and MeIQx-induced hepatocarcinogenesis in male F344 rats

Chemopreventive effects of arctiin, a lignan isolated from Arctium lappa (burdock) seeds, on the initiation or post initiation period of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) induced mammary carcinogenesis in female rats and on 2-amino-3, 8-dimethylimidazo[4,5-f]quinoxaline (MeIQx)-associated hepatocarcinogenesis in male rats were examined. In experiment 1, female Sprague-Dawley (SD) rats were given intragastric doses of 100 mg/kg body wt of PhIP once a week for 8 weeks as initiation. Groups of 20 rats each were treated with 0.2 or 0.02% arctiin during or after PhIP initiation. Control rats were fed 0.2 or 0.02% arctiin, or basal diet alone during the experimental period. Animals were killed at the end of week 48. Although the incidence of mammary carcinomas did not significantly differ among the PhIP-treated groups, multiplicity was significantly decreased in rats given 0.2 (0.7+/-0.7, P<0.05) or 0.02% (1.0+/-1.1, P<0.05) arctiin after PhIP initiation as compared with the PhIP alone controls (2.1+/-2.5). The average number of colon aberrant crypt foci was also significantly decreased in these two groups. Pancreas acidophilic foci were induced in PhIP treated animals with slight decrease in the multiplicity with arctiin during the initiation phase. For liver carcinogenesis, groups of 15 male F344 rats were given a single intraperitoneal injection of diethylnitrosamine (DEN) and starting 2 weeks later, they were administered 0.03% MeIQx in the diet, MeIQx together with 0.5% arctiin, 0.1% arctiin or basal diet for 6 weeks. They were subjected to two-third partial hepatectomy 3 weeks after DEN initiation and killed at the end of week 8 for glutathione S-transferase placental form (GST-P) immunohistochemistry. The numbers and areas of preneoplastic GST-P positive foci were elevated by the treatment with MeIQx, and further increased by the simultaneous treatment with arctiin. These results indicate that arctiin has a protective effect on PhIP-induced carcinogenesis particularly in the mammary gland in the promotion period. On the other hand, it may have a weak co-carcinogenic influence on MeIQx-induced hepatocarcinogenesis. In addition, the results suggested that PhIP is a weak pancreatic carcinogen in female SD rats, targeting acinar cells.     

Selective induction of glandular stomach carcinoma in F344 rats by N-methyl-N-nitrosourea

To study the carcinogenic action of N-methyl-N-nitrosourea (MNU) on the stomach, MNU in distilled water at a concentration of 400 ppm was provided as drinking water to F344 male rats for 25 weeks (group I) or 15 weeks (group II). Twenty weeks following the cessation of the administration, invasive adenocarcinomas were found in the glandular stomach in 100% of 16 rats in group I and 38% of 21 rats in group II. Bone formation occurred within the stroma of carcinoma in 5 rats in group I. No neoplastic lesions developed in the esophagus, forestomach or duodenum of any rat. Thus, MNU in drinking water selectively induces glandular stomach carcinoma in high incidence in these rats.     

Effect of different levels of dietary trans fat or corn oil on azoxymethane-induced colon carcinogenesis in F344 rats

The effect of various levels of dietary corn oil or trans fat on azoxymethane (AOM; CAS: 25843-45-2)-induced carcinogenesis was investigated in female F344 rats fed the AIN-76 semipurified diets. Starting at 5 weeks of age, groups of rats were fed the low-fat diet containing 5% corn oil (designated as low-fat control diet). At 7 weeks of age, all animals except the vehicle-treated controls, were given sc injections of AOM (15 mg/kg body wt, once weekly) for 3 weeks. After 1 week, groups of animals were transferred to semipurified diets containing 13.6% corn oil and 23.5% corn oil or high-fat diets containing 5.9% corn oil plus 5.9% trans fat plus 11.8% Oleinate (low trans fat), 5.9% corn oil plus 11.8% trans fat plus 5.9% Oleinate (intermediate trans fat), and 5.9% corn oil plus 17.6% trans fat (high trans fat). Fecal bile acids were measured in vehicle-treated rats. All animals were necropsied 34 weeks after the last AOM injection. The animals fed the 23.5% corn oil diet had a higher incidence of colon tumors than did those in the groups fed the 5 and 13.6% corn oil diets. There was no difference in colon tumor incidence between the 5 and 13.6% corn oil diet groups. The animals fed the high-fat diets containing low trans fat, intermediate trans fat, and high trans fat developed significantly fewer liver and colon tumors and more small intestinal tumors than did the rats fed 23.5% corn oil diet. The excretion of fecal deoxycholic acid, lithocholic acid, and 12-ketolithocholic acid was higher in animals fed the 23.5% corn oil diet compared to the excretion in animals fed the other diets.     

Enhancement of esophageal carcinogenesis in male F344 rats by dietary phenylhexyl isothiocyanate

The purpose of this study was to evaluate the potential effectsof dietary 6-phenylhexyl isothiocyanate (PHITC) on N-nitrosomethylbenzylamine(NMBA)-induced esophageal carcinogenesis in rats. Groups of15 male F344 rats received weekly s.c. injections of NMBA in20% dimethylsulfoxide or the vehicle alone for 15 consecutiveweeks. Two weeks prior to initiation of carcinogen or vehicleinjections rats were provided with modified AIN-76A diet ormodified AIN-76A diet containing PHITC at levels of 0.4, 1.0or 2.5 µmol/g diet. Experimental controls consisted ofgroups that received only the vehicle (vehicle controls), NMBA(carcinogen controls) or PHITC at the high dose level of 2.5µmol/g diet. No esophageal tumors or preneoplastic lesionswere detected in rats that received the vehicle or PHITC alone.In contrast, all rats treated with NMBA alone or PHITC + NMBAexhibited esophageal tumors and preneoplastic esophageal lesions.In groups that received PHITC + NMBA tumor multiplicity wasincreased by 21–69% when compared with rats treated withNMBA alone, indicating that PHITC enhanced esophageal tumorigenesisin this model system. These results, in conjunction with ourprevious work, demonstrate that arylalkyl isothiocyanates mayinhibit or enhance esophageal tumorigenesis in the NMBA-treatedrat. The ability of isothiocyanates to inhibit or enhance experimentaltumorigenesis may depend on alkyl chain length of the isothiocyanate,the animal species examined and the specific carcinogen employed.     

Influence of dietary fat, caloric restriction, and voluntary exercise on N-nitrosomethylurea-induced mammary tumorigenesis in rats

The effect of dietary fat, energy restriction, and exercise on N-nitrosomethylurea (NMU:CAS:684-93-5)-induced mammary tumorigenesis in female F344 rats was investigated. Rats were fed the NIH-07 diet until N-nitrosomethylurea administration on Day 50 of age, when they were transferred to six treatment groups. Three sedentary groups were fed either high-fat (20%, w/w), medium-fat (10%), or low-fat (5%) diets ad libitum (HFAL, MFAL, LFAL, respectively); two sedentary groups were fed high fat and medium fat diets restricted to 75% of the food consumed by their ad libitum counterparts (HFR, MFR), and one group was fed a HFAL diet but allowed free access to an activity wheel (HFEX). Tumor yields among the three ad libitum sedentary groups were significantly greater in the HFAL and MFAL groups when compared to the LFAL group. Dietary restriction reduced tumor yields by more than 90% of ad libitum controls regardless of fat intake. Voluntary exercise reduced tumor yields and delayed time of tumor appearance in HFEX animals to levels similar to those found in LFAL animals. Animals with voluntary access to exercise wheels averaged between 1.03 and 2.85 miles/day, consumed more food (+18%), and exhibited greater weight gain (+13%) than their sedentary counterparts. Restricted animals exhibited significantly decreased body weight gains (-15%) compared to their ad libitum counterparts, but no differences in weight gains were detected among the HFAL, MFAL, and LFAL groups, despite widely varying amounts of fat intake. Body composition studies indicated that body fat content was not influenced by the quantity of fat consumed in the diet, but was significantly reduced by caloric restriction (-20 to 26%) and exercise (-20%). While the precise mechanisms underlying the tumor-promoting effects of HFAL diets and the antipromoting effects of energy restriction and exercise remain to be elucidated, available evidence suggests that these effects are not due to alterations in energy homeostasis per se, but may instead be exerted indirectly, and perhaps independently via endocrine, paracrine, or neurohormonal mechanisms.     

Histopathological and immunohistochemical studies on nickel sulfide-induced tumors in F344 rats

Twenty-five tumors induced in F344 male rats were examined histologically and immunohistochemically using antibodies against myoglobin, myosin, desmin and cathepsin B. Eight were from rats which had been given intramuscular (im) injection and 17 were from rats which had been given subcutaneous (sc) injection of 5 mg of Ni3S2. Among 10 rhabdomyosarcomas, myoglobin was detectable in 3, myosin in 8, and desmin in all, but cathepsin B was present in none. Out of 8 malignant fibrous histiocytomas, cathepsin B was detectable in all, but the other antigens were absent. In a leiomyosarcoma, only desmin was detected. In two fibrosarcomas, none of the markers were detected. In four undetermined tumors, one reacted only with anti-desmin antibody, two with only anti-cathepsin B antibody, and one with none of the antibodies. Of the three myogenic markers utilized in this study, anti-desmin antibody appeared to be the most sensitive. Cathepsin B was found mainly in the histiocytic cells of malignant fibrous histiocytoma. Thus, desmin appears to be particularly valuable in distinguishing immature myogenic tumors from other primitive tumors, while cathepsin B is useful in distinguishing malignant fibrous histiocytoma from other pleomorphic mesenchymal tumors.     

Inhibitory effect of chlorophyllin on PhIP-induced mammary carcinogenesis in female F344 rats

Chlorophyll and chlorophyllin, a water-soluble salt of chlorophyll,have been reported to inhibit carcinogen—DNA binding andexert antimutagenic activity for some carcinogenic heterocyclicamines and aflatoxins. In the present experiment, the possibleinhibitory effects of chlorophyllin on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) carcinogenicity were investigated. FemaleF344 rats were administered both PhIP, 0.02% in the diet, andchlorophyllin, 1% in the diet (group 1), or either PhIP (group2) or chlorophyllin (group 3) alone for 54 weeks. The incidenceof mammary adenocarcinomas induced by PhIP was reduced by chlorophyllinco-administration from 40 % (8/20 rats) to 15% (3/20). Whilethe difference was not statistically significant, the multiplicityof adenocarcinomas was significantly (P < 0.05) reduced bychlorophyllin co-administration from 0.50 per animal to 0.15.On the other hand, incidence of colon adenomas was slightly,but not significantly, increased from 10% to 20%. Neither mammarynor colon adenocarcinomas were observed in group 3. Thus, chlorophyllinreduced PhIP mammary carcinogenesis, suggesting that chlorophyllinis an effective chemopreventor when ingested simultaneouslywith the carcinogen.     

Induction of oral cavity tumors in F344 rats by tobacco-specific nitrosamines and snuff

The tumorigenic activities toward the oral cavity of snuff, its extracts, and two of its major nitrosamines, N'-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were evaluated in male F344 rats. In one protocol, groups of 21-30 rats were treated beginning at age 10 weeks by chronic application to the oral cavity for 131 weeks of either H2O, an H2O extract of snuff, an H2O extract of snuff enriched with ten times its indigenous concentration of NNN and NNK, or with NNN and NNK in H2O. The incidence of oral cavity tumors in the rats treated with NNN and NNK was 8 of 30, compared to 0 of 30 in controls (P less than 0.05). These results demonstrate that NNN and NNK can induce tumors locally in the oral cavity of F344 rats. Oral cavity tumors were also observed in 3 of 30 rats treated with snuff extract enriched with NNN and NNK, but not in the rats treated with snuff extract alone. In a second protocol, a test canal was surgically created in the lower lip of groups of 21-32 rats, and either snuff, H2O-extracted snuff, or snuff enriched with its own H2O extract was inserted in the test canal 5 times weekly for 116 weeks. A group of 10 control rats had surgery only. Among the 32 rats treated with snuff, 3 had oral cavity tumors; one was a squamous cell carcinoma originating in the test canal and invading the gingiva, one was a papilloma of the test canal, and one was a papilloma of the hard palate. Oral cavity tumors were also observed in 2 of 21 rats treated with H2O-extracted snuff and 1 of 32 rats treated with snuff enriched with its H2O extract. Oral tumors were not observed in control rats. The results of this study indicate that snuff and individual nitrosamines present in snuff can induce oral cavity tumors in F344 rats and support the epidemiological observations which indicate that snuff dipping causes oral cancer in man.     

Adrenal regulation of mammary tumorigenesis in female Sprague-Dawley rats: incidence, latency, and yield of mammary tumors

Huggins and Morii (J. Exp. Med., 114: 741, 1961) reported that massive adrenal necrosis occurs in 79 and 100% of female Sprague-Dawley rats receiving 20 and 30 mg, respectively, of the mammary carcinogen 7,12-dimethylbenz(a)anthracene (DMBA). Here, adrenal necrosis and regeneration were studied in 158 rats for up to 21 days post-DMBA by radioautography of the adrenals of animals given 50 microCi [3H]thymidine 30 min before sacrifice. Adrenal cell proliferation was markedly inhibited 21 days post-DMBA. Regenerated adrenals were more susceptible to this adrenocorticolytic effect. To investigate if alterations in adrenal function modify tumorigenesis, animals underwent adrenalectomies (ADX), hypophysectomies, ovariectomies, and pituitary transplants alone or in combination 6 days after receiving DMBA (20 mg/100 g intragastrically) at 50 days of age. To prevent adrenal necrosis, 24 animals were pretreated with metyrapone. Methylprednisolone acetate, 1 mg i.m., was given to 40 animals every 5 days beginning 6 days post-DMBA. There were 50 non-DMBA-treated intact and surgical controls. DMBA was necessary but not sufficient to induce mammary tumors. No tumors developed in controls or in 46 animals hypophysectomized 6 days after DMBA. Metyrapone reduced tumor incidence and yield. ADX after DMBA treatment increased the tumorigenic response and eliminated resistance to tumorigenesis in older rats. Only three tumors developed in DMBA-treated rats receiving methylprednisolone acetate. Mammary tumorigenesis was increased by pituitary transplant 6 days after DMBA to intact and ADX animals. Ovariectomy 6 days after DMBA was as effective as methylprednisolone acetate in preventing tumorigenesis; ADX did not overcome either inhibition. We conclude that adrenal hormones inhibit proliferation of initiated mammary cells.     

Effect of varying proportions of dietary fat on the development of N-nitrosomethylurea-induced rat mammary tumors

The influence of diets varying in their relative proportions of dietary fat on N-nitrosomethylurea (NMU)-induced mammary tumorigenesis, was assessed. Animals were initiated on day 50 of age with 25 mg/kg NMU and then placed on casein-based AIN-76A diets containing 5, 10, 16 and 23% corn oil (wt/wt). There were 30 animals/group and the experiment was terminated 22 weeks post-NMU. It was found that animals fed diets containing 16 and 23% corn oil exhibited tumor incidences between 2 and 3 times that of animals fed diets containing 5 and 10% corn oil. No differences in tumor incidence could be detected between the 5 and 10% groups or the 16 and 23% groups, suggesting that a threshold occurs at some point between 10 and 16% fat (or 20 and 33% of total calories as fat). The results of this study suggest that the tumor-promoting effects of dietary fat are manifested in terms of a threshold, rather than a linear dose-response effect.