Canine Mammary Tumors: Classification, Biomarkers, Traditional and Personalized Therapies.

In recent years, many studies have focused their attention on the dog as a proper animal model for human cancer. In dogs, mammary tumors develop spontaneously, involving a complex interplay between tumor cells and the immune system and revealing several molecular and clinical similarities to human breast cancer. In this review, we summarized the major features of canine mammary tumor, risk factors, and the most important biomarkers used for diagnosis and treatment. Traditional therapy of mammary tumors in dogs includes surgery, which is the first choice, followed by chemotherapy, radiotherapy, or hormonal therapy. However, these therapeutic strategies may not always be sufficient on their own; advancements in understanding cancer mechanisms and the development of innovative treatments offer hope for improved outcomes for oncologic patients. There is still a growing interest in the use of personalized medicine, which should play an irreplaceable role in the research not only in human cancer therapy, but also in veterinary oncology. Moreover, immunotherapy may represent a novel and promising therapeutic option in canine mammary cancers. The study of novel therapeutic approaches is essential for future research in both human and veterinary oncology.

Amounts of tissue magnesium and some trace elements in cats with mammary tumors related to various clinicopathological parameters.

BACKGROUND: Mammary tumors are one of the major malignancies seen in cats. Researchers have indicated the similarity between the epidemiological and clinicopathological patterns of feline mammary tumors and human breast cancer (HBC). In recent years, the investigation of trace elements in cancer tissues becomes prevalent in HBC due to the role of these elements in biochemical and physiological processes. This study, it is aimed to evaluate some trace elements in feline mammary tumors according to clinical and pathological findings. METHODS: A total of 60 tumoral masses from 16 female cats with mammary tumors were included in the study. The study groups were formed according to histopathology as malignant epithelial tumor (MET; n = 39) and hyperplasia and dysplasia (H&D; n = 21). Copper (Cu), Iron (Fe), Magnesium (Mg), Manganese (Mn), Selenium (Se) and Zinc (Zn) trace elements in mammary tissues were analyzed by using an inductively coupled plasma-optical emission spectrophotometer. RESULTS: The mean age and weight of the cats were 11.75 ± 0.75 years and 3.35 ± 0.21 kg; respectively. Eleven of 16 cats were intact whereas the rest of them had been spayed. Metastases were observed in 10 cats. Tissue Mg level in group MET was significantly higher than in group H&D (P < 0.01) while the other elements had not significant differences between the groups. In group MET, analyzed elements were not statistically significant related to the inflammation, ulceration and invasion to the peripheral muscle (P > 0.05). However, tissue Fe level was significantly higher in T2 than in T3 (P < 0.05). The mean levels of tissue Fe, Mg and Mn had significant differences related to histological grading as P < 0.01, P < 0.05 and P < 0.001; respectively. A mild to severe correlation was found between tissue Zn and Se, Cu, Fe, Mg, and Mn levels. CONCLUSION: Tissue Mg and some trace elements were evaluated in feline mammary tumours in regard to various clinicopathological parameters. Tissue Mg level was sufficient to differentiate the malignant epithelial tumors from hyperplasia and dysplasia. However, Mn and Se tended to distinguish different tumor types. Tissue Fe, Mg and Mn had significant differences related to histological grading. Also, the Fe level was significantly higher in T2 than in T3 and Zn level tended to be higher in T3 than in T1. It was concluded that Mg, Se, Mn, Fe, Cu and Zn provided useful information on the pathogenesis of feline mammary tumors. Further research is needed on the tissue and serum concentrations of trace elements which may provide valuable information for the disease prognosis.

Bcl2-dependent antineoplastic effects of Calotropis procera root extract against canine mammary tumor cells.

There has been a prevailing trend in the application of herbal medicine as cancer therapeutics. Calotropis procera is an ayurvedic plant applied to ameliorate various illnesses. There is no report on the anti-tumor effects of the root of the plant on canine tumors, although it has been used for the treatment of various diseases in human medicine. The objective of the present study was to investigate the antitumor potential of ethanolic root extract of C. procera against canine mammary tumor cell line (CF41-Mg). MTT, western blot, and flow cytometry assays were carried out to evaluate the possible cytotoxicity and apoptosis induction of the extract. MTT results showed that the extract had a potent cytotoxic activity in a dose-dependent manner with an IC50 of 9.00 µg mL-1. Based on the results of flow cytometry and western blotting, IC50 concentration of the extract induced significant apoptosis in the studied cell line, possibly through down-regulation of Bcl-2 expression. The results of the present study clearly indicated that the root extract of C. procera had promising anti-cancer activity and could be considered as a candidate for the treatment of mammary tumors.

Dietary Selenium Supplementation Does Not Attenuate Mammary Tumorigenesis-Mediated Bone Loss in Male MMTV-PyMT Mice.

Bone wasting occurs during the progression of breast cancer and contributes to breast cancer mortality. We evaluated the effect of methylseleninic acid (MSeA), an anti-carcinogenic form of selenium, on bone microstructural changes in the presence of mammary tumors in a male breast cancer model of mouse mammary tumor virus-polyomavirus middle T-antigen (MMTV-PyMT). In this study, we performed microcomputed tomographic analysis of femurs and vertebrae collected from a study showing that dietary supplementation with MSeA reduces mammary tumorigenesis in male mice. Compared to age-matched, non-tumor-bearing mice (MMTV-PyMT negative), the presence of mammary tumors significantly reduced the bone volume fraction, trabecular thickness, and bone mineral density while it increased the structure model index in femurs, but not in vertebrae. Moreover, mammary tumorigenesis decreased plasma concentrations of osteocalcin. Supplementation with MSeA did not affect these changes in MMTV-PyMT mice. In conclusion, mammary tumorigenesis caused bone loss in MMTV-PyMT mice. However, dietary supplementation with MSeA did not attenuate mammary tumor-associated bone loss in this model of male breast cancer.

Nelumbo nucifera Leaves Prevent NMU-Induced Mammary Tumor through Downregulation of Fatty Acid Synthase, Estrogen Receptor-α and Her2 Expression.

Diet polyphenol can reportedly prevent the formation of breast-cancer cells. Nelumbo nucifera leaf extract (NLE) is enriched with polyphenols and has several cellular functions, such as anti-atherosclerosis, anti-inflammation, and antitumor. In this study, we investigated the role of NLE in the prevention of N-methyl-N-nitrosourea (NMU)-induced mammary tumor formation. Cotreatment with NLE significantly reduced the NMU-induced tumor incidence, number, and volume. NLE administration significantly repressed the tumor growth and weight of nude mice upon inoculation with BT-474 cancer cells. Immunohistochemical staining indicated that fatty acid synthetase, estrogen receptor (ER)-α, and phosphorylated ER-α were obviously reduced in the cancer part of BT-474 inoculated nude mice upon administration of 2% NLE. Western blot analysis revealed that NLE and NLPE (polyphenol-rich NLE) repressed ER-α expression and phosphorylation and decreased the phosphorylation of Her-2 without affecting their expression. Overall, NLE and NLPE exhibited more effective antitumor abilities in NMU-induced mammary cancer formation than with tamoxifen and Herceptin.

Rodent Models Assessing Mammary Tumor Prevention by Soy or Soy Isoflavones.

While epidemiological studies performed in Asian countries generally show that high levels of dietary soy are associated with reduced breast cancer risk, studies in Western countries have typically failed to show this correlation. In an attempt to model the preventative actions of soy on mammary tumor development, rodent models have been employed. Thirty-four studies were identified that evaluated the impact of soy products or purified soy isoflavones on mammary tumor initiation (studies evaluating established mammary tumors or mammary tumor cell lines were not included) and these studies were separated into mammary tumors induced by chemical carcinogens or transgenic expression of oncogenes based on the timing of soy administration. Regardless of when soy-based diets or purified isoflavones were administered, no consistent protective effects were observed in either carcinogen-induced or oncogene-induced mammary tumors. While some studies demonstrated that soy or purified isoflavones could reduce mammary tumor incidence, other studies showed either no effect or tumor promoting effects of soy products or isoflavones. Most importantly, only five studies found a decrease in mammary tumor incidence and six studies observed a decrease in tumor multiplicity, two relevant measures of the tumor preventative effects of soy or isoflavones. The variable outcomes of the studies examined were not completely surprising given that few studies employed the same experimental design. Future studies should be carefully designed to more accurately emulate soy consumption observed in Asian cultures including lifetime exposure to less refined soy products and potentially the incorporation of multigenerational feeding studies.

Long-chain omega-3 polyunsaturated fatty acids decrease mammary tumor growth, multiorgan metastasis and enhance survival.

Epidemiological studies show a reduced risk of breast cancer (BC) in women consuming high levels of long-chain (LC) omega-3 (ω-3) fatty acids (FAs) compared with women who consumed low levels. However, the regulatory and mechanistic roles of dietary ω-6 and LC-ω-3 FAs on tumor progression, metastasis and survival are poorly understood. Female BALB/c mice (10-week old) were pair-fed with a diet containing ω-3 or an isocaloric, isolipidic ω-6 diet for 16 weeks prior to the orthotopic implantation of 4T1 mammary tumor cells. Major outcomes studied included: mammary tumor growth, survival analysis, and metastases analyses in multiple organs including pulmonary, hepatic, bone, cardiac, renal, ovarian, and contralateral MG (CMG). The dietary regulation of the tumor microenvironment was evaluated in mice autopsied on day-35 post tumor injection. In mice fed the ω-3 containing diet, there was a significant delay in tumor initiation and prolonged survival relative to the ω-6 diet-fed group. The tumor size on day 35 post tumor injection in the ω-3 group was 50% smaller and the frequencies of pulmonary and bone metastases were significantly lower relative to the ω-6 group. Similarly, the incidence/frequencies and/or size of cardiac, renal, ovarian metastases were significantly lower in mice fed the ω-3 diet. The analyses of the tumor microenvironment showed that tumors in the ω-3 group had significantly lower numbers of proliferating tumor cells (Ki67+)/high power field (HPF), and higher numbers of apoptotic tumor cells (TUNEL+)/HPF, lower neo-vascularization (CD31+ vessels/HPF), infiltration by neutrophil elastase+ cells, and macrophages (F4/80+) relative to the tumors from the ω-6 group. Further, in tumors from the ω-3 diet-fed mice, T-cell infiltration was 102% higher resulting in a neutrophil to T-lymphocyte ratio (NLR) that was 76% lower (p < 0.05). Direct correlations were observed between NLR with tumor size and T-cell infiltration with the number of apoptotic tumor cells. qRT-PCR analysis revealed that tumor IL10 mRNA levels were significantly higher (six-fold) in the tumors from mice fed the ω-3 diet and inversely correlated with the tumor size. Our data suggest that dietary LC-ω-3FAs modulates the mammary tumor microenvironment slowing tumor growth, and reducing metastases to both common and less preferential organs resulting in prolonged survival. The surrogate analyses undertaken support a mechanism of action by dietary LC-ω-3FAs that includes, but is not limited to decreased infiltration by myeloid cells (neutrophils and macrophages), an increase in CD3+ lymphocyte infiltration and IL10 associated anti-inflammatory activity.

Antineoplastic effect of pectic polysaccharides from green sweet pepper (Capsicum annuum) on mammary tumor cells in vivo and in vitro.

The present study investigated the antineoplastic effects of pectic polysaccharides that were extracted from green sweet pepper (Capsicum annuum [CAP]) in the Ehrlich carcinoma in mice and in human mammary tumor lineages. After the subcutaneous inoculation of 2 × 106 Ehrlich tumor cells, Female Swiss mice received 50, 100, or 150 mg/kg CAP or vehicle orally once daily or methotrexate (2.5 mg/kg, i.p., every 5 days) for 21 days. CAP dose-dependently reduced Ehrlich tumor growth. It also reduced the viability of MCF-7, MDA-MB-231, and MDA-MB-436 human mammary cell lineages. Treatment with CAP reduced the gene expression of vascular endothelial growth factor in vivo and in vitro, reduced vessel areas of the tumors, and induced necrosis in Ehrlich solid tumors. CAP treatment significantly increased Interleukin-6 in tumors. The antineoplastic effect of CAP appears to depend on the regulation of inflammation and angiogenesis. Further studies are encouraged to better understand the CAP potential for the treatment of breast tumors.

Chemoprevention of LA7-Induced Mammary Tumor Growth by SM6Met, a Well-Characterized Cyclopia Extract.

Breast cancer (BC) is the leading cause of cancer-related deaths in women. Chemoprevention of BC by using plant extracts is gaining attention. SM6Met, a well-characterized extract of Cyclopia subternata with reported selective estrogen receptor subtype activity, has shown tumor suppressive effects in a chemically induced BC model in rats, which is known to be estrogen responsive. However, there is no information on the estrogen sensitivity of the relatively new orthotopic model of LA7 cell-induced mammary tumors. In the present study, the potential chemopreventative and side-effect profile of SM6Met on LA7 cell-induced tumor growth was evaluated, as was the effects of 17ß-estradiol and standard-of-care (SOC) endocrine therapies, such as tamoxifen (TAM), letrozole (LET), and fulvestrant (FUL). Tumor growth was observed in the tumor-vehicle control group until day 10 post tumor induction, which declined afterward on days 12-14. SM6Met suppressed tumor growth to the same extent as TAM, while LET, but not FUL, also showed substantial anti-tumor effects. Short-term 17ß-estradiol treatment reduced tumor volume on days prior to day 10, whereas tumor promoting effects were observed during long-term treatment, which was especially evident at later time points. Marked elevation in serum markers of liver injury, which was further supported by histological evaluation, was observed in the vehicle-treated tumor control, TAM, LET, and long-term 17ß-estradiol treatment groups. Alterations in the lipid profiles were also observed in the 17ß-estradiol treatment groups. In contrast, SM6Met did not augment the increase in serum levels of liver injury biomarkers caused by tumor induction and no effect was observed on lipid profiles. In summary, the results from the current study demonstrate the chemopreventative effect of SM6Met on mammary tumor growth, which was comparable to that of TAM, without eliciting the negative side-effects observed with this SOC endocrine therapy. Furthermore, the results of this study also showed some responsiveness of LA7-induced tumors to estrogen and SOC endocrine therapies. Thus, this model may be useful in evaluating potential endocrine therapies for hormone responsive BC.

Oridonin inhibits aberrant AKT activation in breast cancer.

Aberrant activation of phosphatidylinosito-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) signaling in cancer has led to pursuit of inhibitors for targeting this pathway. However, inhibitors of PI3K and AKT have failed to yield efficacious results without adverse effects. Here, we screened a library containing 441 authenticated traditional chinese medicine (TCM) plant extracts by examining their effect on cell viability of a human mammary epithelial cell line HMEC-PIK3CAH1047R, which expresses mutant PIK3CAH1047R and has constitutively active AKT signaling. We found that Oridonin, an extract from Rabdosia rubescens, reduced cell viability to the greatest extent. Oridonin binds to AKT1 and potentially functions as an ATP-competitive AKT inhibitor. Importantly, Oridonin selectively impaired tumor growth of human breast cancer cells with hyperactivation of PI3K/AKT signaling. Moreover, Oridonin prevented the initiation of mouse mammary tumors driven by PIK3CAH1047R. Our results suggest that Oridonin may serve as a potent and durable therapeutic agent for the treatment of breast cancers with hyperactivation of PI3K/AKT signaling.

Marine fish oil is more potent than plant-based n-3 polyunsaturated fatty acids in the prevention of mammary tumors.

Marine-derived n-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have been shown to inhibit mammary carcinogenesis. However, evidence regarding plant-based α-linolenic acid (ALA), the major n-3 PUFA in the Western diet, remains equivocal. The objective of this study was to examine the effect of lifelong exposure to plant- or marine-derived n-3 PUFAs on pubertal mammary gland and tumor development in MMTV-neu(ndl)-YD5 mice. It is hypothesized that lifelong exposure to n-3 PUFA reduces terminal end buds during puberty leading to delayed tumor onset, volume and multiplicity. It is further hypothesized that plant-derived n-3 PUFAs will exert dose-dependent effects. Harems of MMTV-FVB males were bred with wild-type females and fed either a (1) 10% safflower (10% SF, n-6 PUFA, control), (2) 10% flaxseed (10% FS), (3) 7% safflower plus 3% flaxseed (3% FS) or (4) 7% safflower plus 3% menhaden (3% FO) diet. Female offspring were maintained on parental diets. Compared to SF, 10% FS and 3% FO reduced (P<.05) terminal end buds at 6 weeks and tumor volume and multiplicity at 20 weeks. A dose-dependent reduction of tumor volume and multiplicity was observed in mice fed 3% and 10% FS. Antitumorigenic effects were associated with altered HER2, pHER-2, pAkt and Ki-67 protein expression. Compared to 10% SF, 3% FO significantly down-regulated expression of genes involved in eicosanoid synthesis and inflammation. From this, it can be estimated that ALA was 1/8 as potent as EPA+DHA. Thus, marine-derived n-3 PUFAs have greater potency versus plant-based n-3 PUFAs.

Dampness-Heat Accelerates DMBA-Induced Mammary Tumors in Rats.

OBJECTIVE: To investigate the impact of dampness-heat (DH) on the development of mammary tumors in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rats. METHODS: Forty rats were randomly divided into 3 groups in a randomized block design, including the control group (n=13), DMBA group (n=14), and DMBA plus DH group (n=13). Rats in the DMBA group and DMBA plus DH group were intragastrically administrated with DMBA (100 mg/kg) for twice, once per week, while rats in the control group were treated with equivalent volumes of sesame oil. After DMBA administration, rats in the DMBA plus DH group were exposed to a simulated climate chamber with ambient temperature (33.0±0.5°C) and humidity (90%±5%) for 8 weeks, 8 h per day. The body weight, time of tumor formation, and number of tumors were measured weekly to calculate tumor incidence, average latency period, average number of tumors, and average tumor weight. At the end of the experiment, the levels of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) in serum, and the contents of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1ß in serum and tumor tissue were measured, respectively. Some tumor tissues were processed for hematoxylin-eosin staining to determine the histopathological changes. RESULTS: Compared with DMBA, DMBA plus DH significantly increased the average number of tumors, average tumor weight, levels of serum MMP-9, TIMP-1, TNF-α and IL-1ß, and contents of tumor tissue TNF-α and IL-1ß (P<0.05 or P<0.01). CONCLUSION: DH could accelerate the development of mammary tumors through increasing the expressions of MMP-9, TIMP-1, TNF-α and IL-1ß in DMBA-induced rats.

A Comparative Study on the In Vitro Effects of the DNA Methyltransferase Inhibitor 5-Azacytidine (5-AzaC) in Breast/Mammary Cancer of Different Mammalian Species.

Murine models are indispensible for the study of human breast cancer, but they have limitations: tumors arising spontaneously in humans must be induced in mice, and long-term follow up is limited by the short life span of rodents. In contrast, dogs and cats develop mammary tumors spontaneously and are relatively long-lived. This study examines the effects of the DNA methyltransferase (DNMT) inhibitor 5-Azacytidine (5-AzaC) on normal and tumoral mammary cell lines derived from dogs, cats and humans, as proof of concept that small companion animals are useful models of human breast cancer. Our findings show that treatment with 5-AzaC reduces in vitro tumorigenicity in all three species based on growth and invasion assays, mitochondrial activity and susceptibility to apoptosis. Interestingly, we found that the effects of 5-AzaC on gene expression varied not only between the different species but also between different tumoral cell lines within the same species, and confirmed the correlation between loss of methylation in a specific gene promotor region and increased expression of the associated gene using bisulfite sequencing. In addition, treatment with a high dose of 5-AzaC was toxic to tumoral, but not healthy, mammary cell lines from all species, indicating this drug has therapeutic potential. Importantly, we confirmed these results in primary malignant cells isolated from canine and feline adenocarcinomas. The similarities observed between the three species suggest dogs and cats can be useful models for the study of human breast cancer and the pre-clinical evaluation of novel therapeutics.

Selenium Induces an Anti-tumor Effect Via Inhibiting Intratumoral Angiogenesis in a Mouse Model of Transplanted Canine Mammary Tumor Cells.

Selenium (Se) has been widely reported to possess anti-tumor effects. Angiogenesis is the formation of new blood vessels and is required to supply oxygen, nutrients, and growth factors for tumor growth, progression, and metastasis. To explore whether the anti-tumor effect of Se was associated with angiogenesis in vivo, we studied the effects of sodium selenite (Sel) and methylseleninic acid (MSA) on tumors induced by canine mammary tumor cells (CMT1211) in mice; cyclophosphamide (CTX) served as a positive control. The results showed that the Se content was significantly increased in the Sel and MSA groups. Se significantly inhibited the tumor weights and volumes. Large necrotic areas and scattered and abnormal small necrotic areas were observed in the Se treatment group. Immunofluorescence double staining showed a reduction in the microvessel density (MVD) and increment in the vessel maturation index (VMI) compared with the untreated control group. As expected, the protein and mRNA levels of the angiogenesis factors angiopoietin-2 (Ang-2), platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) were decreased in the Se-treated tumors by IHC, as shown by western blotting and RT-QPCR. We also found that organic Se MSA provided stronger inhibition of tumor growth compared with inorganic sodium selenite (Sel). Altogether, our results indicated that Se exerted anti-tumor effects in vivo at least partially by inhibiting angiogenic factors.

Anestesia convencional e técnica de tumescência em cadelas submetidas à mastectomia. Avaliação da dor pós-operatória / Conventional anesthesia and tumescent technique in bitches which underwent mastectomy. Evaluation of postoperative pain

As neoplasias mamárias são as mais comuns em cadelas e geralmente acometem fêmeas de meia-idade a idosas, não castradas ou que foram submetidas ao procedimento de ovario-histerectomia tardiamente. A principal forma de tratamento é a excisão cirúrgica, sendo a ressecção unilateral das glândulas mamárias o procedimento mais realizado na prática veterinária. O objetivo do presente estudo foi comparar a dor pós-operatória em cadelas submetidas ao procedimento de mastectomia unilateral sob efeito das técnicas anestésicas de tumescência ou convencional. Foram utilizadas 20 cadelas, divididas em dois grupos: um grupo recebeu apenas a anestesia geral inalatória (grupo convencional), e o outro recebeu adicionalmente a anestesia infiltrativa por tumescência (grupo tumescência). Elas foram avaliadas nas primeiras 24 horas do período pós-operatório, e o processo álgico foi avaliado pela escala de dor da Universidade de Melbourne. As médias dos escores obtidos ao longo do tempo foram submetidas ao teste de Tukey a um nível de 5 por cento de significância (p<0,05). Não foram observadas diferenças significativas entre os grupos com relação à dor pós-operatória. A técnica anestésica infiltrativa por tumescência apresentou a vantagem da redução do sangramento transoperatório e mostrou-se exequível em pequenos animais, entretanto seu uso está relacionado à experiência e à preferência do cirurgião e do anestesista...

Mammary tumors are the most common neoplasm in bitches. Intact, mild to advanced aged female dogs are generally more affected. The main treatment is surgical excision and unilateral mastectomy is the most performed procedure in veterinary practice. The aim of this study was to compare the postoperative pain in dogs which underwent unilateral mastectomy and were anesthetized with tumescence and conventional anesthesia. Twenty bitches were randomly divided into two groups: Conventional Group (GC), which received only general inhalational anesthesia and Tumescence Group (GT), which also received tumescence anesthesia. All dogs were evaluated during the first 24 hours postoperatively. The scale of Pain from the University of Melbourne was used for the evaluation. The tukey test at a 5 percent level of significance (p <0.05) was used. There were no significant differences between groups related to postoperative pain. The technique of tumescent anesthesia reduced bleeding during surgery and is feasible in dogs. Its use is related to the surgeon and anesthesiologist's experience and preferences...

Effect of Antioxidants on the Incidence of 7, 12-dimethylbenzanthracene-induced Mammary Tumor in Rats

The inhibitory effect of selenium, vitamin E, and BHA on DMBA-induced mammary tumorigenesis in rats was investigated. Dietary vitamin E (200 IU/Kg diet) alone could not reduce the tumor incidence at 25 weeks after DMBA administration (10mg DMBA/rat) when selenium was deficient. Selenium supplementation (2ppm in drinking water) to rats fed a practical diet (0.17 ppm Se) reduced the tumor incidence to 14.3% from 75% at 27 weeks after DMBA administration. Dietary supplementation of BHA (0.75%) also reduced the incidence of DMBA-induced mammary tumor to 42.9% at 27 weeks after DMBA-treatment. Rats fed a diet deficient in both selenium and vitamin E contained significantly lower glutathione peroxidase activity and higher malondialdehyde in muscle. However, supplementation of selenium or BHA to the rats fed a practical diet did not alter the malondialdehyde content and glutathione peroxidase activities in muscle, skin and mammary gland. Dietary selenium increased the tissue selenium level. DMBA-induced mammary tumorigenesis was reduced by antioxidants tested but the anticarcinogenic effect of selenium or BHA seems to be independent of glutathione peroxi-dase activity.