Search Results (82)

Breast cancer remains a leading cause of cancer-related mortality worldwide, highlighting the need for novel therapeutic strategies that selectively target tumor cells while minimizing systemic toxicity. Dietary ammonium hydroxide enhancement (AHE) has previously been shown to modulate metabolic pathways in animal studies. However, its potential as a localized anticancer therapy has not been investigated. In the present study, we evaluated the antitumor efficacy and systemic safety of NH₄OH using complementary in vitro and orthotopic breast cancer xenograft models. MDA-MB-231 breast cancer cells and non-tumorigenic MCF10A mammary epithelial cells were treated with increasing concentrations of NH₄OH (2.5–225 µM) to assess dose-dependent effects on cell proliferation, viability, and apoptosis. Following this, MDA-MB-231 cells were orthotopically xenografted into female athymic nude mice and treated by intratumoral injection with NH₄OH using a stepwise dose-escalation regimen (0.01%, 0.1%, and 0.5%; total volume of 20 μL per tumor divided between two injection sites) or phosphate-buffered saline (PBS). Differences between treatment groups of mammary tumors and kidney tissues were analyzed molecularly and histologically. NH₄OH significantly suppressed MDA-MB-231 cell growth and metabolic activity, with minimal effects on MCF10A cells, and induced apoptosis in MDA-MB-231 cells without detectable apoptotic induction in MCF10A cells. In vivo, although tumor volume only showed a non-significant downward trend, histological and molecular analyses demonstrated substantial alterations in tumor biology. NH₄OH treatment induced molecular changes consistent with an antitumor response, including increased Caspase-3 and p53 expression, reduced BCL2 and Ki-67 expression, and attenuation of TNFα, IL-6, and TLR4 inflammatory signaling. Furthermore, the tumor architecture in T-NH tumors displayed increased pale eosinophilic regions and reduced cellular density, suggestive of treatment-associated tumor tissue disruption. Histological analysis of kidney tissue showed no evidence of overt renal toxicity. Indeed, localized NH₄OH administration was associated with reduced renal inflammatory and apoptotic signaling, preserved renal morphology, and increased expression of the ammonia transporters RHBG and RHCG. Cross-sectional morphometric measurements showed decreased area for the distal convoluted tubules in NH₄OH-treated samples. Although this initial preclinical study was limited by a relatively small sample size, further studies are warranted to validate these findings and define the molecular mechanisms underlying NH₄OH-mediated antitumor activity. Collectively, these findings suggest that intratumoral NH₄OH modulates tumor metabolic, inflammatory, and apoptotic pathways associated with a less aggressive tumor phenotype while showing no overt molecular or histological evidence of renal injury, supporting further investigation as a localized metabolic intervention targeting molecular and histological drivers of breast cancer progression. Full article

Background/Objectives: Mammary adipose tissue functions as an active endocrine organ, potentially influencing breast carcinogenesis through metabolic and inflammatory mechanisms. This systematic review synthesized evidence on associations between mammary adipocyte size, obesity-related adipose tissue dysfunction, and breast cancer. Methods: A comprehensive literature search was conducted across MEDLINE, EMBASE, CENTRAL, CINAHL and Web of Science (January 2010–September 2025) for observational studies measuring mammary adipocyte size in human breast tissue. Two reviewers independently conducted screening following the Cochrane Review’s rigorous methodology, and bias assessment using the ROBINS-E (Risk Of Bias In Non-randomized Studies of Exposure) tool. Random-effects meta-analyses were performed for quantifiable outcomes, with heterogeneity assessed using the I² statistic. Results: Twenty-one studies were included in the systematic review. Meta-analyses indicated that mammary adipocyte diameter was positively correlated with body mass index (8 studies; n = 720; correlation coefficient (r) = 0.44, 95% CI: 0.29–0.56), crown-like structure (CLS) density (5 studies; n = 383; r = 0.45, 95% confidence interval (CI): 0.36–0.54), aromatase expression (3 studies; n = 333; r = 0.36, 95% CI: 0.05–0.61), and three studies compared adipocyte diameter between CLS-positive and CLS-negative tissue (n = 297; mean difference = 9.69 μm, 95% CI: 4.94–14.44). Conclusions: This review summarizes moderate positive correlations of mammary adipocyte size with systemic adiposity, local breast inflammation, and aromatase expression, supporting a potential role for mammary adipose tissue in obesity-related metabolic dysfunction. Future prospective studies with standardized measurement protocols and improved control for confounding variables are needed to strengthen causal inference and clinical applications. Full article

Background: Comparative oncology uses naturally occurring cancers in companion animals to study tumor biology and therapeutic responses relevant to human cancer. Spontaneous feline tumors are increasingly recognized as useful comparative models because they arise in immunocompetent hosts, develop under shared environmental exposures, and can reproduce selected clinical, histopathologic, molecular, and therapeutic features of human malignancies. Methods: This review compares feline oral squamous cell carcinoma (FOSCC) with human head and neck squamous cell carcinoma (HNSCC), and feline mammary carcinoma (FMC) with human breast cancer, emphasizing shared pathologic, molecular, tumor microenvironment, and therapeutic features. Results: Recent immunohistochemical, genomic, transcriptomic, and biomarker studies have identified shared features between feline and human cancers. FOSCC resembles human HNSCC through aggressive local invasion, histologic features, therapeutic resistance, and recurrent alterations of TP53, MYC, and PTEN. FMC shows strong overlap with aggressive human triple-negative breast cancer, including reduced hormone receptor expression, recurrent TP53, PIK3CA, and CXCL12/CXCR4 signaling alterations, and tumor microenvironment features involving immune-checkpoint, inflammatory, and angiogenic pathways. FOSCC clinical trials and emerging clinical investigations into FMC treatments further support the use of cats for translational therapy evaluation. Conclusions: FOSCC and FMC are promising comparative oncology models for human HNSCC and aggressive breast cancer, respectively. Future multicenter studies incorporating standardized tumor classification and grading, predefined stratification criteria, and clinically meaningful endpoints will be essential to strengthen their translational value. Full article

Although human mammary glands were traditionally considered sterile, accumulating evidence has established the presence of distinct microbial communities that may have colonized breast tissue primarily via retrograde nipple flow or via hematogenous or lymphatic translocation from other body sites. Comparative studies reveal differences in the microbiota of healthy and diseased breast tissues, with variations in microbial signatures across breast cancer subtypes and in comparison with adjacent normal tissues. This review synthesizes current evidence on the composition of the breast microbiome, the factors shaping its development, and alterations it undergoes in inflammatory and malignant breast diseases. Furthermore, the article discusses mechanistic insights, methodological challenges, and future therapeutic perspectives based on published studies employing culture-independent approaches, such as 16S rRNA gene sequencing and metagenomic analyses. Key host-related factors influencing breast-associated microbial communities, including hormonal regulation, environmental exposure, diet, and therapeutic interventions, are explored. The existing literature is assessed to identify key associations between the breast microbiome and host signaling pathways, as well as the significant challenges that remain unresolved, including low biomass contamination, inter-study variability, limited longitudinal data, and an incomplete understanding of causality. Addressing these limitations is critical for advancing microbiome-based diagnostic and therapeutic strategies for breast disease. Full article

Background/Objectives: Histone deacetylase (HDAC) inhibitors have been shown to prime the response to immunotherapy (IMT) treatment by inducing immune activation and infiltration to target tumor cells. Many studies primarily focus on adaptive immune cells and their expression of pro-inflammatory markers, like somatostatin receptor 2 (SSTR2); however, macrophages are known to help mediate key tumor microenvironment changes. The goal of this study is to evaluate the effects of HDAC inhibitors and IMT on macrophages, their expression of SSTR2, and their impact on the treatment response in triple-negative breast cancer (TNBC). Methods: Cytotoxic effects of HDAC inhibitors on 4T1 mouse mammary carcinoma cells, including suberoylanilide hydroxamic acid (SAHA), were evaluated using flow cytometry. Bone marrow-derived macrophages (BMDMs) were stimulated to M1-like and M2-like phenotypes and treated with SAHA to explore the effects on SSTR2 expression in different macrophage phenotypes. 4T1-tumor-bearing BALB/c mice were used to evaluate the therapy response to four treatments: saline control, SAHA, anti-PD-1 + anti-CTLA-4 checkpoint blockade IMT, or a combination of SAHA + IMT. Additional cohorts of 4T1-tumor-bearing BALB/c mice and NOD SCID mice, which lack adaptive immune cells, were euthanized for early evaluation of tumor-associated macrophage (TAM) populations via flow cytometry and cytokine analysis. One-way independent ANOVAs and log-rank tests were used to compare group differences. Results: SAHA promotes SSTR2 expression on M1-like BMDMs in vitro. SAHA promotes M2-like TAMs in vivo and stimulates pro-inflammatory, anti-tumor cytokine production in combination with IMT. Conclusions: SAHA drives SSTR2 expression and anti-tumor innate immune responses with additive effects in combination with immunotherapy in preclinical TNBC. Full article

Staphylococcus aureus (S. aureus)-induced mastitis poses a significant threat to animal husbandry. This condition triggers sustained mammary inflammation, oxidative stress, and disrupts mitochondrial homeostasis, ultimately impairing mammary gland function and milk yield. Conjugated linoleic acid (CLA) is a long-chain fatty acid found in meat and dairy products derived from ruminants. It exhibits multiple biological activities, including anti-cancer, anti-inflammatory, and antioxidative stress-alleviating effects. Thus, this study sought to determine whether CLA alleviates S. aureus-induced mastitis in Hu sheep through the PPARG-UCP2 axis. Fifteen lactating Hu sheep were randomly allocated into three groups (n = 5): control group, model group, and CLA group. The CLA group received 1 mg/mammary gland of CLA via intramammary infusion for seven days, followed by S. aureus challenge (5 × 10⁷ cells/mL, 2 mL/mammary gland) in the model and CLA groups, while the control group received saline. Venous blood and mammary tissue samples were collected at two days post-infection. The results demonstrated that S. aureus infection significantly upregulated the expression of inflammatory factors (IL-1β, IL-6, and NF-κB) in the mammary tissue of Hu sheep, p < 0.01. Relative to the control, the model group showed increased ROS and MDA levels, a diminished NAD+/NADH ratio, and downregulated expression of the antioxidant factors SOD, Nrf2, HO-1, and SIRT3, p < 0.01. Furthermore, the expression of p-AMPK and mitophagy-related factors (PARKIN, PINK1, and LC3b) showed a statistically significant increase in the model group than in the control group, p < 0.01. S. aureus infection also suppressed the expression of PPARG and UCP2, p < 0.01. In contrast, the CLA group showed lower levels of inflammatory factors (IL-1β, IL-6, and NF-κB), ROS and MDA, while the NAD+/NADH ratio and the expression of antioxidant factors (SOD, p-Nrf2, HO-1, and SIRT3) were elevated compared with the model group, p < 0.01. Moreover, the expression of p-AMPK and mitophagy-related factors (PARKIN, PINK1, and LC3b) was reduced in the CLA group relative to the model group, p < 0.05. Concurrently, the expression of PPARG and UCP2 was higher in the CLA group than in the model group, p < 0.001. These findings demonstrated that S. aureus infection induced mastitis in Hu sheep mammary tissue, whereas CLA alleviated the infection by upregulating the PPARG-UCP2 pathway, thereby reducing inflammation, oxidative stress, and mitophagy levels. This study offers a novel perspective on mammary tissue repair during mastitis and expands the understanding of UCP2’s biological role. Full article

Reduced lactation after forced weaning induces a tumor-promoting environment in the mammary gland, triggering key regulatory nodes common to both post-lactation involution and breast cancer, including calpains and miRs. We investigated how lactation duration modulates these nodes using two murine models: short lactation (forced weaning) and prolonged lactation (spontaneous-weaning). Additionally, the role of inflammation in calpain regulation was assessed in forced-weaned NOS-2 knockout mice. Morphological and molecular analysis in mammary tissue included histochemical staining, qPCR, enzymatic activity, Western blot and miRNA-seq. Mammary gland involution after prolonged lactation resulted in milder inflammation, reduced cell death and tissue remodeling, and lower collagen deposition compared to short lactation. The expression, activity and function of calpain-2 was found to be more sensitive to the model of lactation and inflammatory environment compared to calpain-1. Even after full regression (28 days postpartum), prolonged lactation maintained lower calpain-2 levels and higher expression of tumor suppressors miR-10b and miR-143/145. These persistent molecular differences suggest spontaneous weaning as the optimal model for healthy mammary gland regression, whereas forced weaning sustains alterations in calpain-2 and regulatory miRs that may increase post-partum breast cancer risk. The potential long-term influence of lactation duration on breast cancer development warrants further consideration in both clinical and public health contexts. Full article

Coumarin derivatives, whether natural or synthetic, have attracted considerable interest from medicinal chemists due to their versatile biological properties. Their appealing pharmacological activities—such as anticancer, anti-inflammatory, neuroprotective, anticoagulant, and antioxidant effects—combined with the ease of their synthesis and the ability to introduce chemical modifications at multiple positions have made them a widely explored class of compounds. In the scientific literature, there are many examples. On the other hand, dithiocarbamates, originally employed as pesticides and fungicides in agriculture, have recently emerged as potential therapeutic agents for the treatment of serious diseases such as cancer and microbial infections. Moreover, dithiocarbamates bearing diverse organic functionalities have demonstrated significant antifungal properties against resistant phytopathogenic fungi, presenting a promising approach to combat the growing global issue of fungal resistance. Dithiocarbamates linked to coumarin derivatives have been shown to exhibit cytotoxic activity against various human cancer cell lines, including MGC-803 (gastric), MCF-7 (breast), PC-3 (prostate), EC-109 (esophageal), H460 (non-small cell lung), HCCLM-7 (hepatocellular carcinoma), HeLa (cervical carcinoma), MDA-MB-435S (mammary adenocarcinoma), SW480 (colon carcinoma), and Hep-2 (laryngeal carcinoma). Numerous studies have revealed that the inclusion of a dithiocarbamate moiety can provide central nervous system (CNS) activity, particularly through inhibitory potency and selectivity toward acetylcholinesterase (AChE) and monoamine oxidases (MAO-A and MAO-B). Recently, it has been reported that coumarin–dithiocarbamate derivatives exhibit α-glucosidase inhibitory effects and also possess promising antimicrobial activity. This study presents an overview of recent progress in the chemistry of coumarin–dithiocarbamate derivatives, with a focus on their biological activity. Previous review papers focused on coumarin derivatives as multitarget compounds for neurodegenerative diseases and described various types of compounds, with dithiocarbamate derivatives representing only a small part of them. Our work deals exclusively with coumarin dithiocarbamates and their biological activity. Full article

Macrophage-mediated inflammation is known to be involved in the epithelial–mesenchymal transition (EMT) of various types of cancer. This makes macrophage-derived inflammatory factors prime targets for the development of new treatments. This study uncovers the therapeutic potential and action mechanism of DAB-2-28, a small-molecule derived from para-aminobenzoic acid, in the treatment of breast cancer. The luminal MCF-7 and the triple-negative MDA-MB-231 cancer cell lines used in this study represent, respectively, breast cancers in which the differentiation states are related to the epithelial phenotype of the mammary gland and breast cancers expressing a highly aggressive mesenchymal phenotype. In MCF-7 cells, soluble factors from macrophage-conditioned media (CM-MØ) induce a characteristic morphology of mesenchymal cells with an upregulated expression of Snail1, a mesenchymal marker, as opposed to a decrease in the expression of E-cadherin, an epithelial marker. DAB-2-28 does not affect the differential expression of Snail1 and E-cadherin in response to CM-MØ, but negatively impacts other hallmarks of EMT by decreasing invasion and migration capacities, in addition to MMP9 expression and gelatinase activity, in both MCF-7 and MDA-MB-231 cells. Moreover, DAB-2-28 inhibits the phosphorylation of key pro-EMT transcriptional factors, such as NFκB, STAT3, SMAD2, CREB, and/or AKT proteins, in breast cancer cells exposed to different EMT inducers. Overall, our study provides evidence suggesting that inhibition of EMT initiation or maintenance is a key mechanism by which DAB-2-28 can exert anti-tumoral effects in breast cancer cells. Full article

Nuclear receptors are involved in multiple biological processes, among which RORγ can regulate the expression of inflammation-related genes and is thus frequently used as a therapeutic target for cancer. Canine mammary cancer is one of the most common tumor diseases in dogs, with a relative incidence rate of 46.71% for CMT in China over the past five years, severely threatening the life and health of dogs. Therefore, the search for novel drugs targeting canine mammary cancer is of great significance. This study aims to investigate how the RORγ inhibitors W6134 and XY018 affect the expression of inflammatory genes through histone modifications in CMT-N7 cells. These results show that W6134 and XY018 can upregulate signaling pathways related to inflammation and apoptosis and influence the expression of associated genes. The close link between RORγ and inflammation-related genes further confirms that RORγ may serve as a therapeutic target for canine cancer. Additionally, ChIP-qPCR was used to detect the enrichment of histone markers such as P300, H3K27ac, H3K4me1, H3K9la, and H3K9bhb at the target loci of CXCL10 and MECOM genes. Collectively, our findings provide molecular evidence for the protective role of RORγ in canine mammary cancer, potentially by regulating inflammatory pathways via histone modifications, offering new insights for improving the cure rate and survival of affected dogs. Full article

Obesity and metabolic dysfunction are established risk factors for luminal breast cancer, yet current preclinical models inadequately recapitulate the complex metabolic and immune interactions driving tumorigenesis. To develop and characterize an immunocompetent rat model of luminal breast cancer induced by chronic exposure to a cafeteria diet mimicking Western obesogenic nutrition, female rats were fed a cafeteria diet or standard chow from weaning. Metabolic parameters, plasma biomarkers (including leptin, insulin, IGF-1, adiponectin, and estrone), mammary gland histology, tumor incidence, and gene expression profiles were longitudinally evaluated. Gene expression was assessed by PCR arrays and qPCR. A subgroup underwent dietary reversal to assess the reversibility of molecular alterations. Cafeteria diet induced significant obesity (mean weight 426.76 g vs. 263.09 g controls, p < 0.001) and increased leptin levels without altering insulin, IGF-1, or inflammatory markers. Histological analysis showed increased ductal ectasia and benign lesions, with earlier fibroadenoma and luminal carcinoma development in diet-fed rats. Tumors exhibited luminal phenotype, low Ki67, and elevated PAI-1 expression. Gene expression alterations were time point specific and revealed early downregulation of ID1 and COX2, followed by upregulation of MMP2, THBS1, TWIST1, and PAI-1. Short-term dietary reversal normalized several gene expression changes. Overall tumor incidence was modest (~12%), reflecting early tumor-promoting microenvironmental changes rather than aggressive carcinogenesis. This immunocompetent cafeteria diet rat model recapitulates key metabolic, histological, and molecular features of obesity-associated luminal breast cancer and offers a valuable platform for studying early tumorigenic mechanisms and prevention strategies without carcinogen-induced confounders. Full article

PiRNAs are a subclass of non-coding RNAs, 26–31 nucleotides (nt) in length, that form regulatory complexes through their interaction with PIWI proteins. Studies in model organisms have demonstrated that piRNAs play crucial roles in tissue development and in predicting disease outcomes, positioning them as promising targets for developmental regulation and therapeutic intervention. In contrast, research on piRNAs in animal husbandry is still in its early stages and has not received sufficient attention. Despite this, the few studies available in livestock research have revealed that piRNAs serve as key regulators of reproductive development, underscoring their significant regulatory potential in farm animals and justifying further investigation. Accordingly, this review uses the bovine mammary gland as an exemplary case to summarize the progress in piRNA research related to mammary development and disease. The role of piRNAs in regulating breast cancer stem cell proliferation and modulating inflammatory progression is a highly active area of research. We hypothesize that piRNAs may play a potential role in regulating both mammary gland development and mastitis, making them promising targets for enhancing mammary development and overall health in dairy cattle and providing a theoretical foundation for further piRNA applications in animal husbandry. Full article

Background/Objective: Breast cancer remains the most common malignancy among women worldwide, contributing to high morbidity and mortality rates. Many anti-cancer drugs have been derived from medicinal plants, and frankincense from Boswellia carterii is notable for its anti-inflammatory, anti-neoplastic, and anti-carcinogenic properties. Using gas chromatography/mass spectrometry (GC/MS), 48 components were identified in B. carterii essential oil, and the major constituent was α-pinene (35.81%). Method: In this study, we investigated the anti-tumor effects of frankincense essential oil (FEO) and its nano-formulation with chitosan (FEO-CSNPs) using in vitro breast cancer models (MCF-7, MDA-MB-231, and 4T1 cells) and in vivo mouse mammary carcinoma (4T1) models (Balb/c). Results: The results showed significant reductions in cell viability. At 10 μg/mL, the FEO showed the highest reduction in the C-166 cells, while at 100 μg/mL, the FEO exhibited a stronger cytotoxicity in the MDA-MB-231 and 4T1 cells compared to the FEO-CSNPs and CSNPs. The FEO-CSNPs exhibited cell growth arrest in the S, G2/M, and G1/S phases in the MCF-7, MDA-MB-231, and 4T1 cell lines (36.91%, 23.12%, and 33.58%), in addition to increased apoptosis rates in the MCF-7, MDA-MB-231, and 4T1 cell lines (33.04%, 36.39%, and 42.19%). The wound healing assays revealed a decreased migratory ability in the treated cells. The in vivo experiments in the balb/c mice demonstrated a reduction in the tumor volume, with a histopathological analysis confirming extensive tumor necrosis. Moreover, the FEO and FEO-CSNPs showed notable antioxidant and arginase activity. The gene expression analysis via qPCR indicated the upregulation of tumor suppressor genes and the downregulation of oncogenes. Conclusions: These findings suggest that FEO and its nano-formulation, particularly in the form of FEO-CSNPs as an oral formulation, display enhanced efficacy, warranting further preclinical and clinical research to develop innovative treatment strategies. Full article

Breast cancer is the most common and the leading cause of cancer death in women worldwide; treating invasive breast carcinomas is challenging due to the side effects of chemotherapeutics. Compounds isolated from natural sources have been proposed as potential molecules for cancer therapy; for instance, the homoisoflavonoid brazilin has shown pharmacological properties, including anti-tumoral and anti-inflammatory activities. In this study, we isolated brazilin from the heartwood of Haematoxylum brasiletto; then, we performed a semi-synthesis by adding three methyl or acetyl groups to the core structure of brazilin. We confirmed the identity of brazilin and its derivatives by spectroscopic data (¹H NMR and ¹³C NMR) and measured their purity by optical rotation. Then, we analyzed the effects of brazilin and its derivatives in three mammary gland-derived cell lines: the TNBC MDA-MB-231, the ERα(+) MCF7, and the non-tumorigenic MCF10A. We evaluated the cell viability by MTT assays, cell migration by wound-healing assays, and focal adhesion kinase (FAK) activation by Western blot. Regarding biological assays, the MTT assay showed that these compounds showed cytotoxic effects on the MCF7 and MDA-MB-231 breast cancer cells at 20 µM but was not toxic in non-tumorigenic MCF10A mammary epithelial cells. Specifically, the greatest effects found from treatment with the compounds were in the MDA-MB-231 cell line, where the IC50 of brazilin was 49.92 μM, and for MCF7, the brazilin-(OAc)₃ was 49.97 μM. These effects were dose- and time-dependent, as well as being associated with a decrease in the levels of cell migration and FAK activation. Full article

Early detection and comprehensive diagnostic approaches for breast cancer are essential for improving prognosis. When it comes to changes in the skin of the breast or the nipple–areola complex (NAC), particularly if they are unilateral, it is essential to be vigilant, as these changes could be an early sign of underlying malignancy or other pathologies. Primary breast malignancies, such as mammary Paget’s disease (MPD), can manifest as erythema, scaling, or ulceration of the NAC, while secondary cutaneous metastases from other breast carcinomas may present as nodules, erythematous plaques, or inflammatory reactions. Non-malignant inflammatory conditions, including eczema or mastitis, can also mimic these changes; histologic evaluation is the gold-standard diagnostic tool. The usefulness of conventional diagnostic techniques breast lesions has been confirmed, but in recent years, reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) have emerged as additional tools to diagnose cases characterized by cutaneous changes; they may, therefore, result in new perspectives on the non-invasive diagnosis of MPD. RCM is a non-invasive diagnostic technique that allows high-resolution images of the skin at microscopic level in real time, offering a promising approach to the non-invasive diagnosis of MPD, particularly when a lesion is not clinically evident and may mimic other benign or inflammatory conditions. We describe an atypical clinical presentation of mammary Paget’s disease diagnosed early by reflectance confocal microscopy evaluation and confirmed histologically. Full article