Hormonal profiles and estrogen receptors in Egyptian female breast cancer patients.

AIMS AND BACKGROUND: Hormones are considered to be an important factor in the etiology of breast cancer. Serum hormonal profiles of premenopausal and postmenopausal breast cancer patients as well as estrogen receptor (ER) concentrations in breast cancer tissues were examined in an attempt to establish a possible association between hormones and breast cancer risk and to elucidate the biological features of the disease among Egyptian female patients. METHODS: Levels of estradiol (E2), testosterone (T), progesterone (P), LH, FSH, prolactin, T3, T4 and TSH were measured by highly specific radioimmunoassays in the sera of women with breast cancer and compared to those of control subjects. ER concentrations in breast tumor tissues were measured using 125I-radioreceptor assay. RESULTS: Levels of T and prolactin showed a significant increase in both premenopausal and postmenopausal patients. E2 and P levels were significantly increased in follicular premenopausal and postmenopausal patients. Luteal E2 showed non-significant changes, whereas the luteal P level was significantly decreased. No significant alterations were found in the levels of serum LH, FSH, T3, T4 and TSH either in premenopausal or postmenopausal patients. Higher levels of ER were found in the tumors of postmenopausal than in those of premenopausal patients. A positive correlation was found between levels of ER and age of the patients (r = 0.35), whereas a negative correlation was observed between ER and serum E2 (r = -0.26). CONCLUSIONS: This study provides evidence of an association between high levels of serum E2 and T and increased risk of breast cancer in postmenopausal women. Abnormalities in serum P and prolactin are probably associated with a breast cancer risk and ER may be considered as a biochemical marker for breast cancer development.

Effect of Vicia faba and bran feeding on nitrosamine carcinogenesis and formation.

The goal of this work was to study the effect of the most common Egyptian food items, Vicia faba beans (VF) and bran, on the carcinogenicity of dibutylnitrosamine (DBN) precursors (dibutylamine and nitrite). Mice receiving DBN precursors showed a delayed gain in body weight as well as decreased protein level and 5-nucleotidase activity. Acid ribonuclease, alkaline phosphatase, and DNA level and rate of synthesis were significantly increased compared with corresponding controls. Hepatomas and bladder papillomas developed in 60% and 40% of mice, respectively, after nine months of treatment. On the other hand, administration of VF or bran, in addition to DBN precursors, lessened the damage caused by DBN precursors alone, except DNA level and rate of synthesis were elevated. Alkaline phosphatase was also elevated when bran was administered with DBN precursors. However, these elevations were still less than corresponding elevations in mice receiving DBN precursors alone. The incidence of hepatoma was also reduced to only 20% for both groups. Meanwhile, incidence of bladder papillomas was only 20% in mice receiving VF in addition to DBN precursors, and bladder papillomas were completely absent in mice receiving bran in addition to DBN precursors. In vitro studies were also performed to clarify the effect of VF or bran on diphenylnitrosamine (DPhNA) and its formation from its precursors (diphenylamine and nitrite). The study revealed that VF and bran have the ability to eliminate nitrite and DPhNA from the reaction media and to reduce the rate of formation of DPhNA from its precursors. This reaction depends on the concentration and form of VF or bran and the duration of the reaction. Thus it is concluded that some naturally occurring food items, such as VF and bran, could protect humans against the hazardous effect of nitrosamines and their precursors.

Effect of soybean, Vicia faba, and vitamin C on the carcinogenicity of DMBA.

A single dose of 10 mg of 7,12-dimethylbenz[a]anthracene (DMBA), administered to rats through intragastric intubation, was sufficient to induce many biochemical and histopathological changes in their mammary tissue. Significant increases were observed in the activity levels of the enzymes acid ribonuclease, 5-nucleotidase, alkaline phosphatase, and beta-glucuronidase in mammary tissue homogenates of DMBA-treated rats after an experimental period of five months. Histopathological studies of the mammary tissue also revealed malignant epithelial tumors (cribriform carcinoma) induced among 85% of the treated rats, with an incidence of 4 tumors in 12 mammary glands. Nevertheless, administration of 30% soybean in the diet of rats or 5,000 ppm ascorbic acid in their drinking water in addition to DMBA revealed a significant chemoprotective effect against the carcinogenesis induced by DMBA alone. This chemoprotective effect was demonstrated by the normalization of the activity levels of the enzymes studied in mammary tissue homogenates, because most of the enzymes were maintained at near the levels in the control animals. The incidence and number of tumors were also decreased. Cribriform carcinoma was observed in 50% of the rats, and the incidence of the affected glands was 2 in 12 mammary glands among both groups. On the other hand, a less chemoprotective effect was observed due to Vicia faba administration.

Distribution and toxicity of 5-fluorouracil after intraperitoneal and anal submucosal administration.

The correlation of 5-fluorouracil (5-FU) distribution and its toxicity had been investigated in Albino rats. H3-5-FU was administrated either by intraperitoneal (I.P.) or anal submucosal (A.sm.) route. 5-FU was promptly distributed in different organs with marked accumulation in the pelvic area after A.sm. and in liver and kidney after I.P. administration. Acute toxicity (L.D. 50) was stronger after I.P. (63 mg/kg compared with 80 mg/kg after A.sm.). Side effects expressed in elevation of transaminase and alkaline phosphatase and increase in liver tissue glucose-6-phosphatase and total white blood count were much pronounced after I.P. administration. The results suggest the possibility of using A.sm. route for administration.

Soybean and ascorbate feeding in experimental carcinogenesis: immunological studies.

Soybean and/or sodium ascorbate have been demonstrated to have protective effects against carcinogenicity of the nitrosamine precursors dibutylamine and sodium nitrite in rats. Some immunological aspects of the cellular and humoral compartments of the immune system were investigated in this experimental model. The parameters chosen for the study were the lymphocyte transformation test, migration inhibition test, the demonstration of Fc and Fc plus complement receptors on the lymphocyte surface (EA and EAC rosette-forming test), and the determination of total serum proteins and their fractions. The results indicated that dibutylnitrosamine can be formed in vivo from its precursors dibutylamine and sodium nitrite, and that it affects the immune system of rats which showed an immunosuppressed state present as depressed cellular and humoral immunity. The protective role of soybean and/or ascorbate against carcinogenicity of dibutylamine and sodium nitrite was demonstrated by the absence of tumor formation. These agents were found to be capable of increasing the defense mechanism of rats by enhancing both cellular and humoral immunity in the presence of the carcinogenic nitrosamine precursors dibutylamine and sodium nitrite.

Potentiation of therapeutic effect of methanesulphonate and protection against its organ cytotoxicity by vitamin C in Ehrlich ascites carcinoma bearing mice.

The effect of vitamin C upon the therapeutic index and side effects produced by methanesulfonate of aminoglycols (drug 864T, NSC 140117) had been evaluated in a laboratory system. The antitumor action of 864T, vitamin C and their combination were evaluated in Ehrlich ascites carcinoma (EAC) cells in vivo. Tissue toxicity was assessed using liver and intestinal DNA, RNA, protein contents and their synthesis as parameters. In addition, G-6-pase, 5-Nase and Alk, pase activity levels in both tissues were also measured. Drug 864T (200 mg/kg) produced 50 percent long-term survivors in tumor bearing mice in addition to 10 percent early mortality while in combination with vitamin C (250 mg/kg x 6), there was 80 percent long term survivors with no mortality related to drug toxicity. No toxicity, in all the parameters used, was observed when 864T was given in combination with vitamin C. Drug 864T alone produced a significant decrease in protein content of both liver and intestinal tissue while in combination with vitamin C normal levels were maintained. In addition, all the parameters used were either elevated or decreased by 864T treatment and returned to normal levels in combination with vitamin C. This study proved that vitamin C may be useful not only to potentiate the effect of anticancer drug 864T on the Ehrlich ascites carcinoma but also to antagonize the side effects of the drug.

Effects of soyabean and ascorbic acid on experimental carcinogenesis.

1. Ultrastructural changes in liver tissue of mice fed nitrosamine precursors, dibutylamine and nitrite, were observed. 2. The protective effect of soyabean in a diet containing nitrosamine precursors was demonstrated. 3. Liver tissue was examined to investigate the anticarcinogenicity of ascorbic acid. 4. The significance of soyabean and ascrobic acid in counteracting the potential hazards due to nitrosamine precursors is discussed.

Effect of soybean feeding on experimental carcinogenesis--III. Carcinogenecity of nitrite and dibutylamine in mice: a histopathological study.

The potential carcinogenic effect of nitrosamine precursors, DBA (dibutylamine) and nitrite, was clearly demonstrated pathologically in the liver and bladder of male Swiss albino mice. Benign tumours were induced in the bladder with an incidence of 40%, and hepatomas were detected in the liver in 27% of the cases. The protective effect of soybean and ascorbic acid, added separately to the diet or to the drinking water respectively, was demonstrated by a marked reduction in dysplastic features and absence of tumour in both the liver and the urinary bladder.

Frequency of active ras oncogenes in human bladder cancers associated with schistosomiasis.

The frequency of active ras oncogenes in human bladder cancers associated with schistosomiasis, the cause of which is suspected to be a chemical carcinogen(s) in urine, was examined. Of 9 squamous cell carcinomas of the bladder surgically obtained in Egypt, none scored as positive in the regular DNA transfection assay using NIH/3T3 cells as recipients. The restriction fragment length polymorphism assay at codon 12 of the H-ras gene confirmed the absence of an activating mutation at this site in all of them. Western blotting analysis of electrophoretic mobilities of the ras p21 proteins, a method which can detect at least some of the point mutations within codons 12 and 61 of ras genes, suggested a point mutation within codon 61 in one out of the 7 tumors analyzed. In contrast to the low frequency of detection of mutationally activated ras oncogenes, enhanced expression of the ras p21 proteins was demonstrated in 4 of them by this analysis. The carcinogenic process involved in the endemic bilharzial bladder cancers is thus not associated with detectable point mutations within ras genes at a higher frequency than those in non-bilharzial bladder cancers in Japan or the USA.

Escherichia coli infection of the urinary bladder: induction of tumours in rats receiving nitrosamine precursors and augmentation of bladder carcinogenesis by N-nitrosobutyl (4-hydroxybutyl)amine.

Experimental introduction of Escherichia coli type 04 into the subserosa of the urinary bladder of female Fischer 344 rats produced chronic bacterial infection in more than 90% of animals. Groups of rats with bacterial infection were given sodium nitrate and either piperazine (Group 1) or dibutylamine (Group 2) in the drinking-water. Control, noninfected animals received nitrate and either piperazine (Group 3) or dibutylamine (Group 4). At 40 weeks, transitional-cell carcinomas of the bladder were detected in 9/30 rats in Group 1 compared to 0/34 in Group 3 (p less than 0.0005), and in 11/34 rats in Group 2 compared to 0/32 in Group 4 (p less than 0.0003). Early changes were examined by scanning and transmission electron microscopy as well as autoradiography. Preneoplastic liver foci were detected in infected groups of animals receiving amine and nitrate, indicating reabsorption of the carcinogen synthesized in situ to induce distant organ transformation. In another experiment, E. coli infection augmented bladder carcinogenesis by N-nitrosobutyl(4-hydroxybutyl)amine (NBHBA), as indicated by earlier appearance of bladder tumours (six weeks compared to nine weeks) and, after 25 weeks, higher incidences of transitional-cell carcinomas (41/46 compared to 39/53, p less than 0.05), squamous metaplasia (43% compared to 9%, p less than 0.0001), glandular metaplasia (26% compared to 13%, p less than 0.05) and muscle invasion (30% compared to 11%, p less than 0.01) in the E. coli-infected group receiving carcinogen compared to the noninfected group receiving carcinogen, respectively. These results indicate that bacterial infection of the urinary bladder may play a major role in bladder carcinogenesis, both by helping in-situ nitrosamine synthesis and by augmenting carcinogenesis by nitrosamines.

Anal submucosal injection: a new route for drug administration in pelvic malignancies. Part I. Experimental study of misonidazole distribution in serum and tissues, with special reference to urinary bladder. Preliminary report.

The anal and oral administration routes were compared in 30 rats to study the distribution of misonidazole, a radiation sensitizer, in the serum and tissues with special reference to the urinary bladder. 14C-labelled misonidazole was administered in a dose of 0.2 ml water/100 gm body weight containing 1 mu Ci misonidazole. The dose was given orally by stomach tube in 15 rats, and was injected in the submucosa of the anal canal in another 15. Animals were then killed after 15, 30, 60, or 120 minutes or after 24 hours. Organs were dissected, and radioactivity was determined in each by the internal standard method. The study has shown that the highest drug concentration in the bladder tissue relative to the serum was achieved with the anal submucosal route. Its level was eight and five times that of the serum 15 and 30 minutes after administration, respectively, in contrast to the oral route in which the drug concentration was one-quarter and equal to the serum level at the same time intervals. The anal route would thus provide the adequate channel required for misonidazole to promote radiation responsiveness in bladder carcinoma.

Serum estrogen level in Egyptian breast cancer patients.

In menstruating 20-29 year old breast cancer patients, the total estrogen level showed a significant increase in the early follicular phase compared to normal healthy subjects. Such a difference was not observed in 30-45 year-old patients. A nonsignificant decrease was observed in the estradiol level of premenopausal breast cancer patients compared to normal healthy subjects. However, in postmenopausal breast cancer patients, the total estrogen level as well as the estradiol level showed a significant increase compared to that of normal healthy subjects.

Immunoglobulin A, G and E levels in Egyptians with cancer: influence of schistosomiasis.

The main patient series consisted of 415 Egyptians attending the Cairo Cancer Institute and comprising 286 bladder cancer, 97 breast cancer, 14 head and neck cancer and 18 gastrointestinal cancer cases. Also included in the study were 36 patients with active schistosomiasis and 89 health controls. Serum IgA, IgG, IgG subclasses, IgE, Schistosoma and Ascaris-specific IgE (RAST) and the acute-phase protein CRP were measured in all, or sub-sets, of the main patient group. The well-established increase in IgE and IgG levels, and the more recently reported increase in the levels of IgG3 and IgG4 subclasses in patients with schistosomiasis, were also found in bilharzial bladder cancer, indicating that humoral immunity persists in cancer-bearing patients. However, the plasma protein profile in bilharzial bladder cancer is further modified by significant increases in the levels of IgA, IgG, IgG1, IgG2 and CRP when compared to levels in patients with Bilharzia in the absence of neoplastic change. Patients with cancers not associated with parasitic infestation also had significant increases in their serum levels of IgG1, IgG2, IgG, IgE and CRP when compared to healthy Egyptian controls, but 41% of these non-bladder cancer patients showed IgE responses to previous parasitic infestations suggesting that any immunological response to cancer would be on the background of a variable non-specific increase of IgE.

Effect of new anticancer methanesulfonates on different biochemical parameters of mouse liver.

The comparative effect to two chemically related new synthesized anticancer drugs namely 3,3'-(methylimino) di(l-propanol)dimethanesulfonate(ester)diphenyl disulfonate, No. 838 D (NSC, 140-115) and 3,3' iminodi-l-propanol dimethanesulfonate(ester)p-toluene disulfonate, No. 864 T (NSC, 140-117), on liver DNA, RNA, lipids and glucose-6-phosphatase, after single 100 mg i.p. injection was studied. Both drugs significantly reduced DNA, RNA and total lipid content on day one and two after administration. Normal levels were regained on day three. Both drugs did not cause a decrease in glucose-6-phosphatase activity level, on the contrary, the enzyme was significantly elevated on day three after administration. The present results demonstrate that these drugs induce no severe or permanent damage to hepatic cells in the administered dose.

A study on the etiological factors of bilharzial bladder cancer in Egypt. 6. The possible role of urinary bacteria.

A correlation was obtained between a positive nitrite test in urine and the severity of urinary bacterial infection. Bacteria isolated from the urine of bilharzial or bladder cancer patients were found to be rich in nitrate reductase activity. "Escherichia coli" was the most common microorganism isolated from these specimens. Urine and several urinary constituents activate bacterial nitrate reductase. beta-Glucuronidase activity in the urine of patients with chronic "Schistosoma haematobium" infection and bladder cancer was measured and shown to be significantly greater than that of urine of normal control subjects. Urinary bacterial infection was shown to be the source of the increased urinary level of enzyme activity at pH 7.0.

A study on the etiological factors of bilharzial bladder cancer in Egypt: 4. beta-carotene and vitamin A level in serum.

The possible role of vitamin A in the pathogenesis of bilharzial bladder cancer among Egyptians, particularly as it relates to the histopathologic tumor type, was investigated. Bilharzial patients and bladder cancer patients with squamous cell carcinoma, the most prevalent type in Egypt, showed significantly lower levels of vitamin A than normal male subjects. In contrast bladder cancer patients with transitional cell carcinoma had levels that were not significantly different from normal male subjects. The possible role of vitamin A in the etiology of bilharzial bladder cancer is discussed.

A study on the etiological factors of bilharzial bladder cancer in Egypt: 5-urinary nitrite in a rural population.

Urinary nitrite was present in 5.6% of 2379 individuals from a rural population infested with "Schistosoma haematobium". A higher frequency was observed in symptomatic patients with active bilharzial cystitis (25%) and patients with bladder cancer associated with schistosomiasis (66.2%); conversely, urinary nitrite was absent in normal urban individuals. The frequency of urinary nitrite was higher in females (6.4%) than males (4.6%), and was more frequent in adults than extremes of age. The presence of urinary nitrite was associated with urinary infection and was commonly accompanied by cellular atypia in urine, in the form of dysplasia. Under these circumstances, carcinogenic nitrosamines are liable to be produced in the bladder from urinary nitrite and amines. These observation support the possible role of urinary bacterial infection, commonly associated with bilharzial cystitis, in bladder carcinogenesis.

Prostatic carcinoma in Egypt: epidemiology and etiology.

In Egypt, the relative frequency of prostatic carcinoma is much lower than elsewhere reported. On the other hand, carcinoma of the male breast shows a strikingly higher frequency. These findings could be related to the prevalent bilharzial infestation which impairs the liver detoxication mechanism. As a result there is an increased estrogen and diminished androgen level. These metabolic changes can contribute to the rarity of prostatic cancer and the high frequency of breast cancer in males.