Calcium and carcinogenesis of the mammary gland.

Effects of dietary calcium on mammary carcinogenesis in rats were investigated because of evidence that calcium counteracts the promotion of colon cancer by dietary fat and because experimental diets for rats normally contain higher amounts of calcium and vitamin D than do human diets. Our earlier experiments indicated that yields of tumors induced in young, Sprague-Dawley rats by 7,12-dimethylbenz(a)-anthracene (DMBA) were higher when dietary calcium, phosphate, and vitamin D were decreased. Results of an experiment in which dietary amounts of calcium, phosphate, and vitamin D were varied independently suggested that phosphate and vitamin D have interactive effects with calcium. Another experiment in which dietary vitamin D alone was varied provided evidence that higher amounts inhibited tumorigenesis in the presence of low amounts of calcium and phosphate but the results with a high-calcium and -phosphate diet were inconclusive. The findings suggest that low amounts of dietary calcium and vitamin D and high amounts of phosphate increase susceptibility to DMBA-induced mammary neoplasia.

Effects of dietary fat, calcium, and vitamin D on growth and mammary tumorigenesis induced by 7,12-dimethylbenz(a)anthracene in female Sprague-Dawley rats.

This study was designed to test the influence of dietary calcium and vitamin D levels on the promotional effect of high-fat diets on chemically induced mammary carcinogenesis. In a small preliminary experiment (Experiment A), 40 female Sprague-Dawley rats, 43 days old, were randomly divided into 5 groups (8 rats/group) and fed a semipurified diet containing 3% sunflower seed oil (SF) by weight, 1.5 mg of calcium/kcal and 0.5 IU vitamin D/kcal of diet. After 1 week, each rat was given 2.5 mg of dimethylbenz(a)anthracene by gastric gavage. One week later, the animals were switched to 1 of 4 diets varying in fat (3 or 20% SF by weight), calcium (1.5 or 0.25 mg/kcal), vitamin D (0.5 or 0.05 IU/kcal), and phosphate or to a fifth diet containing 3% SF by weight, 0.1 mg of calcium/kcal and 0.05 IU of vitamin D/kcal. In all 5 diets, calcium:phosphate weight ratios were maintained at 1.2:1. In animals fed the high-fat diet, reduction of dietary calcium (1.5 to 0.25 mg/kcal) and vitamin D (0.5 to 0.05 IU/kcal) increased the incidence of mammary lesions from 37 to 75% and the total number of lesions from 4 to 16. A trend toward an increase in lesion weight and total lesion burden was also seen. To confirm these results, the experiment was repeated using the same protocol; 126 rats were divided into 6 groups, treated with dimethylbenz(a)anthracene, and fed the diets as described. A sixth diet was included that contained 20% SF by weight, 0.01 mg of calcium/kcal, and 0.05 IU of vitamin D/kcal. As for Experiment A, in animals fed the high-fat diet, reduction of dietary calcium (1.5 to 0.25 mg/kcal) and vitamin D (0.5 to 0.05 IU/kcal) resulted in an increase in total mammary lesions from 31 to 55, a significant increase in average lesion burden/rat with lesions (1.6 +/- 0.6 to 12 +/- 3 g), and a trend toward increasing weight of lesions. The effect was less obvious in animals fed the low-fat diet where, in both experiments, an increase in the incidence of mammary lesions was observed only when the dietary calcium was reduced from 1.5 to 0.1 mg/kcal. These data suggest that decreasing calcium and vitamin D increase the promoting effects of a high-fat diet on mammary tumorigenesis in the rat.

Effects of dietary fat on long-term growth and mammary tumorigenesis in female Sprague-Dawley rats given a low dose of DMBA.

The effects of dietary fat on experimental mammary cancer have typically been observed in relatively young animals. However, in human populations, breast cancer incidence and mortality are highest in postmenopausal women. To develop an animal model that stimulates the human situation more closely, female Sprague-Dawley rats were given a relatively small dose (1.5 mg) of 7,12-dimethylbenz[a]anthracene (DMBA) at 50 days of age while on a semipurified diet containing 3% sunflower-seed oil. One week later, half of the 70 rats were transferred to a diet containing 20% sunflower-seed oil. Very few mammary lesions appeared until about 35 weeks after administration of DMBA, at which time palpable mammary nodules began to appear in many of the animals on the high-fat diet. More than half of the animals in this group had developed nodules by Week 41, whereas the other half of the animals on the low-fat diet developed nodules by Week 46. Rats on the high-fat diet gradually became much more obese than those on the low-fat diet and were significantly heavier at the time they developed lesions. The incidence of nodules continued to increase in both groups and reached 100% in the group fed the high-fat diet by Week 55, with a 70% incidence of adenocarcinomas. At this time, 79% of the animals on the low-fat diet had palpable nodules without a plateau in incidence being reached. On autopsy, adenocarcinomas were found in 57% of animals on the low-fat diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Membrane glycoprotein and surface free energy changes in hypoxic fibroblast cells.

Hypoxia affects the biochemistry of mammalian cells and thus alters their sensitivity to subsequent chemo- and radiotherapy. When V79 Chinese hamster lung fibroblasts were grown under conditions of extreme hypoxia (less than 10 ppm O2) there was a significant shift in the membrane glycoprotein composition. Scanning electron microscopy revealed altered cell surface morphology including loss of pseudopodial projections. Experiments to determine changes in interfacial free energy of these cells using equilibrium two phase systems of poly(ethylene glycol) (PEG) and dextran were carried out. Test fluid droplets of the denser dextran-rich phase were formed on layers of cells in the PEG-rich phase as the bathing medium, and the contact angles the droplets made with the cell layers were measured from photomicrographs. The contact angles on cells in the plateau phase increased significantly with time of exposure to hypoxia, from 25 degrees (zero time) to 35 degrees (6 h) to 60 degrees (9 h). Contact angles on cells in the exponential phase increased from 80 degrees (zero time) to 150 degrees after 20 h of hypoxia. It appears that the altered contact angles reflect changes in cell surface hydrophobicity that may, in part, reflect alterations in the membrane glycoprotein composition.

Changes in ultrastructure and function of hypoxic V79 fibroblast cells after treatment with misonidazole.

Morphologic and enzymatic changes due to exposure to the radiosensitizing chemical, misonidazole, have been identified in V79 cells grown in a system in which oxygen tensions and culture density have been controlled. Misonidazole prevented the increase in mitochondrial size normally seen during exposure of these cells to conditions of moderate hypoxia (2 X 10(3) ppm O2). Mitochondrial size was also significantly decreased in cells from exponential cultures exposed to 1 mmol/l misonidazole. Morphologic changes to the mitochondria that varied from data reported elsewhere were also noted. Misonidazole caused a significant initial decrease in cytochrome oxidase activity after 4 hours of exposure of aerobic and moderately hypoxic cultures that did not return to normal in chronically hypoxic cells during continued exposure to the drug.

Ultrastructural changes in V79 hamster lung fibroblasts during hypoxic exposure.

Cells in tumors that are deprived of their blood supply become hypoxic. These stressed cells adapt to their new environments by altering their metabolic regimen which in time induces cellular structure changes. The morphologic make-up of these O2-deprived cells is the focal point of this electron microscopy study. V-79 hamster lung fibroblast cells grown as monolayer cultures were examined under controlled culture density and oxygen tensions - normal aerobia (2.1 X 10(5) ppm O2), and extreme hypoxia (less than 10 ppm O2). Electron micrographs of these cells demonstrated a loss of structural mitochondrial integrity accompanied with large increases in both mitochondrial and lipid vacuole size following exposure to extreme hypoxia. When these cells were reoxygenated, those mitochondria which had not become degenerate returned to their normal state however, lipids still continued to accumulate in vacuoles for a further 6 h. Addition of 1 mM palmitic acid to aerobic cultures evoked similar lipid and mitochondrial irregularities as were observed in hypoxic cells although, the latter were not as marked. When this saturated fatty acid was added to hypoxic cells no further structural alterations were seen. The cellular changes manifested during this study were subjected to quantitative measurements and these results have given an insight into the scope and variety of ultrastructural changes which have resulted from exposure of cultured cells to hypoxic conditions.

Dietary fiber and cancer: a supplement for intervention studies.

Dietary fiber is one of several variables being considered in the study of the relationship between diet and cancer. Intervention trials in which dietary fiber is increased are the most direct way of assessing the possible role of fiber in this disease. Two dietary snack products have been developed for use in a fiber intervention study: the high-fiber snack (HFS), which supplies 23 g of dietary fiber per day (mostly from wheat bran) and the low-fiber product (LFS), which provides 3.5 g. Over a 12-week period, 28 volunteers consumed the HFS for 6 weeks and the LFS for 6 weeks. Compliance, as assessed by reports, through recovery of a riboflavin marker in the urine and fecal fiber analysis, was good. The only adverse effects reported were mild abdominal discomfort and gas. Serum ferritin and calcium decreased in some subjects, indicating a need to supplement the products with these essential minerals. Consumption of the snacks did not affect total energy intake or the intake of the nutrients monitored.