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Objective(s): Breast neoplasm is a malignancy that can have a poor prognosis. The PI3K/AKT signaling pathway is frequently activated in various tumor types, including breast cancer, leading to alterations in the tumor microenvironment and radioresistance. Selective inhibition of PI3Kca (p110a) has been considered an alternative approach to overcome radioresistance, owing to concerns surrounding the excessive side effects of pan-PI3K inhibitors tested in clinical trials. This investigation aimed to evaluate the efficacy of co-administering PI3Kca knocking out with radiation therapy in mitigating radioresistance and suppressing tumor growth in the MDA-MB-231 cell line. Materials and Methods: The present investigation utilized the CRISPR/Cas9 technique to induce a knockout of the PI3Kca gene. Subsequently, after 24 hr of transfection, gene expression, cell proliferation, apoptosis rate, and angiogenesis were assessed. Results: We demonstrated that knocking out PI3Kca, in combination with radiation, increased apoptosis, reduced the expression of PI3Kca and AKT1 genes, and decreased cell proliferation. The CAM assay analysis has demonstrated that knocking out the PI3Kca gene and radiotherapy substantially reduced the total vessel network length and the number of junctions. Conclusion: The findings of our investigation indicate that the integration of radiation therapy with PI3Kca yielded enhanced radiosensitivity, leading to a marked retardation of tumor progression and an increased survival rate. [ABSTRACT FROM AUTHOR]

Abstract In order to investigate the changes in the properties of the cell culture solution in the effect of cell synchronization via cell starvation (for 12, 24, and 36 h), a new spiral-interdigital pattern of microelectrode as a biosensor has been proposed. Then, to test its superiority, the results of this spiral-interdigital pattern with the results of the commercial pattern have been compared. The cells were selected from breast cancer standard lines (MDA-MB-231). Changes in CV peaks of the secretions were recorded by the spiral-interdigital pattern, in which increasing the interactive surface with homogenous electric paths had been considered by simulation before fabrication. The results of the simulation and experimental procedures showed a meaningful correlation. The occurrence of CV oxidative peaks at about 0.1–0.4 V and reductive peaks at approximately 0 V in the spiral-interdigital biosensor in the starved MDA-MB-231 cell line has been observed. The starvation situation resembles one that does not cause meaningful cell apoptosis or necrosis, and this method is only used to make the cells synchronized. Also, no peak is observed in normal cell growth conditions. In addition, by using the commercial design of the electrodes, no peak is observed in any of the conditions of normal and synchronized growth of the cells. Therefore, it seems that the observed peaks are caused by the agents that are secreted in the cell culture solution in a synchronized situation. Moreover, the design of the new spiral-interdigital electrode can significantly increase the sensitivity of the sensor to receive these peaks due to more space and a uniform electric field.

A new type of hybrid polymer particles capable of carrying the cytostatic drug doxorubicin and labeled with a gallium compound was prepared. These microparticles consist of a core and a hydrogel shell, which serves as the structural matrix. The shell can be employed to immobilize gallium oxide hydroxide (GaOOH) nanoparticles and the drug, resulting in hybrid beads with sizes of approximately 3.81 ± 0.09 μm. The microparticles exhibit the ability to incorporate a remarkably large amount of doxorubicin, approximately 0.96 mg per 1 mg of the polymeric carrier. Additionally, GaOOH nanoparticles can be deposited within the hydrogel layer at an amount of 0.64 mg per 1 mg of the carrier. These nanoparticles, resembling rice grains with an average size of 593 nm by 155 nm, are located on the surface of the polymer carrier. In vitro studies on breast and colon cancer cell lines revealed a pronounced cytotoxic effect of the hybrid polymer particles loaded with doxorubicin, indicating their potential for cancer therapies. Furthermore, investigations on doping the hybrid particles with the Ga-68 radioisotope demonstrated their potential application in positron emission tomography (PET) imaging. The proposed structures present a promising theranostic platform, where particles could be employed in anticancer therapies while monitoring their accumulation in the body using PET. [ABSTRACT FROM AUTHOR]

In order to investigate the changes in the properties of the cell culture solution in the effect of cell synchronization via cell starvation (for 12, 24, and 36 h), a new spiral-interdigital pattern of microelectrode as a biosensor has been proposed. Then, to test its superiority, the results of this spiral-interdigital pattern with the results of the commercial pattern have been compared. The cells were selected from breast cancer standard lines (MDA-MB-231). Changes in CV peaks of the secretions were recorded by the spiral-interdigital pattern, in which increasing the interactive surface with homogenous electric paths had been considered by simulation before fabrication. The results of the simulation and experimental procedures showed a meaningful correlation. The occurrence of CV oxidative peaks at about 0.1–0.4 V and reductive peaks at approximately 0 V in the spiral-interdigital biosensor in the starved MDA-MB-231 cell line has been observed. The starvation situation resembles one that does not cause meaningful cell apoptosis or necrosis, and this method is only used to make the cells synchronized. Also, no peak is observed in normal cell growth conditions. In addition, by using the commercial design of the electrodes, no peak is observed in any of the conditions of normal and synchronized growth of the cells. Therefore, it seems that the observed peaks are caused by the agents that are secreted in the cell culture solution in a synchronized situation. Moreover, the design of the new spiral-interdigital electrode can significantly increase the sensitivity of the sensor to receive these peaks due to more space and a uniform electric field. [ABSTRACT FROM AUTHOR]

Introduction: Cancer is the second leading cause of death globally, and among all types of cancer, triple-negative breast cancer (TNBC) has one of the worst prognoses. The repeated use of chemotherapeutic drugs often leads to drug resistance, complicating treatment outcomes. This study aimed to investigate the effect of jujube alkaloid-rich extract on drug sensitivity and its antitumor effects on paclitaxel-resistant MDA-MB-231 cells in vitro. Materials and methods: This study utilized the MDA-MB-231 breast cancer cell line and its paclitaxel-resistant variant. Cell viability was measured using the XTT assay, while apoptosis was detected through Annexin-PI assays. Various concentrations of jujube extract, alone and combined with paclitaxel, were assessed for synergistic effects in 2D and 3D culture models. Results: The presence of alkaloids in the jujube seed extract was confirmed via the Dragendorff test, with 0.395 g of alkaloids present in 5 g of jujube seed powder. A combination of paclitaxel and jujube extract exhibited dose-dependent cytotoxic effects on paclitaxel-resistant MDA-MB-231 cells. The combination of jujube extract and paclitaxel significantly decreased IC50 from 541.3 μg/ml to 124.3 μg/ml, and the IC20 decreased from 92.1 μg/ml to 23.9 μg/ml when combined with paclitaxel in 2D culture model. The cytotoxicity of the combination was reduced in the 3D culture model. Apoptosis rates were 0.75% in the control group, 13.89% in the alkaloid-rich extract group, 2.59% in the paclitaxel group, and 42.90% in the combination group. Conclusion: Combining alkaloid-rich jujube seed extract with paclitaxel regenerates the cytotoxicity and apoptotic ability of paclitaxel in the paclitaxel-resistant MDA-MB-231 breast cancer cells in both 2D and 3D culture models. This suggests that such combinations might be a viable strategy to overcome drug resistance in TNBC treatments.

Breast cancer is a prevalent and potentially life-threatening disease that affects women worldwide. Natural products have gained attention as potential anticancer agents due to their fewer side effects, low toxicity, and cost effectiveness compared to traditional chemotherapy drugs. In the current study, the network pharmacology approach was used following a molecular docking study to evaluate the therapeutic potential of N. sativa-derived phytochemicals against breast cancer. Specifically, the study aimed to identify potential anticancer agents targeting key proteins implicated in breast cancer progression. Five proteins (i.e., EGFR, MAPK3, ESR1, MAPK1, and PTGS2) associated with breast cancer were selected as receptor proteins. Fourteen phytochemicals from N. sativa were prioritized based on drug-likeness (DL) and oral bioavailability (OB) parameters (with criteria set at DL > 0.18 and OB > 30%, respectively). Subsequent analysis of gene targets identified 283 overlapping genes primarily related to breast cancer pathogenesis. Ten hub genes were identified through topological analysis based on their significance in the KEGG pathway and GO annotations. Molecular docking revealed strong binding affinities between folic acid, betulinic acid, stigmasterol, and selected receptor proteins. These phytochemicals also demonstrated druggability potential. In vitro experiments in the MDA-MB-231 breast cancer cell line revealed that betulinic acid and stigmasterol significantly reduced cell viability after 24 h of treatment, confirming their anticancer activity. Furthermore, in vivo evaluation using a DMBA-induced rat model showed that betulinic acid and stigmasterol contributed to the significant recovery of cancer markers. This study aimed to explore the mechanisms underlying the anticancer potential of N. sativa phytochemicals against breast cancer, with the ultimate goal of identifying novel therapeutic candidates for future drug development. Overall, these results highlight betulinic acid and stigmasterol as promising candidates to develop novel anticancer agents against breast cancer. The comprehensive approach of this study, which integrates network pharmacology and molecular docking study and its experimental validation, strengthens the evidence supporting the therapeutic benefits of N. sativa-derived phytochemicals in breast cancer treatment, making them promising candidates for the development of novel anticancer agents against breast cancer. [ABSTRACT FROM AUTHOR]

Aims of study are identification, isolation, identification and determination of some type bioactive constituents presents in Sanseveria trifasciata (S.trifasciata) with assessment of the effectiveness of ethyl acetate fraction roots and leaves as antiproliferative agents against breast cancer (MDA-MB-231) cell lines. The roots and leaves of S. trifasciata were macerated in n-hexane for defatting, then the defatted components were extracted using a hot extraction technique with 85% aqueous ethanol, and the extracted parts were fractionated first with chloroform, ethyl acetate, and then n-butanol. A fraction of ethyl acetate is analyzed by analytical high performance liquid chromatography (HPLC) to determine its component. Assessment of its cytotoxic activity was performed using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. for 24 hours, multiple concentrations of extracts are used to treat the breast cancer (MDA-MB-231) cell line. After 72 hours of treatment, media were removed and cell viability was calculated. The qualitative HPLC analysis showed that the ethyl acetate fraction of roots contains apigenin, quercetin, chlorogenic acid, luteolin, catechin, and caffeic acid but did not contain kaempferol. On the other hand, the ethyl acetate fraction of leaves contained apigenin, kaempferol, chlorogenic acid, luteolin, and catechin but does not contain quercetin. A cytotoxic study (MTT assay) showed that the IC50% for roots was 89.83 μg/ml and the IC50% for leaves was 77.6 μg/ml. Ethyl acetate extracts of S. trifasciata from both roots and leaves showed anti-breast cancer cell lines through inhibition of proliferation. However, leaves showed a more inhibitory effect at the lower concentration than roots. [ABSTRACT FROM AUTHOR]

Multifunctional nanocomposites (NC) can greatly enhance therapy outcomes by reducing tumor proliferative potential. We created a novel class of Zn_Mn_CMC_FA_sesamol NC in the current work to combat breast cancer (MDA-MB-231) cells. To understand how zinc (Zn), manganese (Mn), carboxymethylcellulose, and folic acid (FA) interact with sesamol, UV-Visible spectrophotometer and Fourier Transform Infrared spectroscopy were used to analyze the absorption behavior of the synthesized NC. The particle size of NC was confirmed by X-ray diffraction and dynamic light scattering. Scanning electron microscopy was used to assess the morphological features of these NCs. photoluminescence spectrum was used to analyze the optical and electron transition molecules of the sample. In addition to MTT analysis, acridine orange/ethidium bromide (AO/EtBr) analysis of reactive oxygen species (ROS) and nuclear staining with 4′,6-diamidino-2-phenylindole as well as flow cytometry were used to confirm the apoptotic activity of Zn_Mn_CMC_FA_sesamol NC on MDA-MB-231 cells. The results showed significant cytotoxicity, apoptosis induction on AO/EtBr, and increased ROS production in treated cells compared to control cells. The cell cycle analysis revealed that NCs triggered apoptosis and arrested the cell cycle in G0/G1 phases. As a conclusion, the created NC serves as a versatile platform for the successful molecularly targeted chemotherapeutic treatment of cancer.

The N1,N2‐diphenyloxalohydrazide OX(HA)2 chemosensor was synthesized and used as a colorimetric and fluorescent receptor to detect Cu2+ and Fe3+ ions in the presence of other metal ions. A range of physicochemical approaches were utilized to characterize the symmetrical oxalohydrazide‐based receptor, including FT‐IR, 1H and 13C NMR, TGA, and mass spectrometry. According to single crystal X‐ray diffraction investigation, OX(HA)2 crystallizes in monoclinic with space group C2/c. UV‐Vis and fluorescence spectroscopy were used to thoroughly explore OX(HA)2's sensing capabilities towards diverse cations. Using measurements of fluorescence intensity, the binding constant for OX(HA)2 with Cu2+/Fe3+ ions was calculated to be 2.78×103 M−1 and 1.12×103 M−1 with the lower detection limit of 0.43 μM and 0.2 μM respectively. With a 1 : 1 binding mechanism validated by Job's plot, mass spectral analysis, FT‐IR spectra, and density functional theory, the dual‐sensing receptor exhibits remarkable selectivity and sensitivity towards Cu2+ and Fe3+ ions. The average fluorescence lifetimes of OX(HA)2, OX(HA)2+Cu2+, and OX(HA)2+Fe3+ were calculated to be 2.19 ns, 1.73 ns, and 1.26 ns, respectively. This fluorescence lifetime measurement results strongly support the complexation of Cu2+ and Fe3+. When Cu2+ and Fe3+ ions bind with OX(HA)2, they induce the 'Turn‐Off' signal. Then, fluorescence imaging of the OX(HA)2 receptor in MDA‐MB‐231 demonstrated that the receptor holds a lot of promise for usage in bioimaging and their IC50 value for cytotoxicity is 96.96 μg. Furthermore, fluorescence microscopy experiments demonstrate that OX(HA)2 can be utilized as a fluorescent probe for detecting Cu2+ and Fe3+ ions in living cells, making it potentially helpful as an anticancer medication. [ABSTRACT FROM AUTHOR]

A novel class of chalcones were synthesized via the Claisen-Schmidt condensation of 6-chloropyridine-3-carbaldhyde with various ortho, meta, and para substituted acetophenones in the presence of a base and a polar protic solvent. The newly synthesized compounds were characterized by using various spectral techniques 1H NMR, 13C NMR, FT-IR, and mass spectroscopy methods. All newly synthesized compounds were evaluated in vitro for anticancer activity against the Human Breast Cancer Cell Line MDA-MB-231, and antioxidant activity was quantified using the DPPH free radical scavenging technique. The anticancer screening findings revealed that the synthesized compounds 3a, 3d, 3e, 3f, and 3j had GI50 of

Abstract This work compared the metabolic profile of a parental MDA-MB-231 cisplatin-sensitive triple negative breast cancer (TNBC) cell line with that of a derived cisplatin-resistant line, to characterize inherent metabolic adaptations to resistance, as a means for marker and new TNBC therapies discovery. Supported by cytotoxic, microscopic and biochemical characterization of both lines, Nuclear Magnetic Resonance (NMR) metabolomics was employed to characterize cell polar extracts for the two cell lines, as a function of time (0, 24 and 48 h), and identify statistically relevant differences both between sensitive and resistant cells and their time course behavior. Biochemical results revealed a slight increase in activation of the NF-κB pathway and a marked decrease of the ERK signaling pathway in resistant cells. This was accompanied by lower glycolytic and glutaminolytic activities, possibly linked to glutamine being required to increase stemness capacity and, hence, higher survival to cisplatin. The TCA cycle dynamics seemed to be time-dependent, with an apparent activation at 48 h preferentially supported by anaplerotic aromatic amino acids, leucine and lysine. A distinct behavior of leucine, compared to the other branched-chain-amino-acids, suggested the importance of the recognized relationship between leucine and in mTOR-mediated autophagy to increase resistance. Suggested markers of MDA-MB-231 TNBC cisplatin-resistance included higher phosphocreatine/creatine ratios, hypotaurine/taurine–mediated antioxidant protective mechanisms, a generalized marked depletion in nucleotides/nucleosides, and a distinctive pattern of choline compounds. Although the putative hypotheses generated here require biological demonstration, they pave the way to the use of metabolites as markers of cisplatin-resistance in TNBC and as guidance to develop therapies.

This work compared the metabolic profile of a parental MDA-MB-231 cisplatin-sensitive triple negative breast cancer (TNBC) cell line with that of a derived cisplatin-resistant line, to characterize inherent metabolic adaptations to resistance, as a means for marker and new TNBC therapies discovery. Supported by cytotoxic, microscopic and biochemical characterization of both lines, Nuclear Magnetic Resonance (NMR) metabolomics was employed to characterize cell polar extracts for the two cell lines, as a function of time (0, 24 and 48 h), and identify statistically relevant differences both between sensitive and resistant cells and their time course behavior. Biochemical results revealed a slight increase in activation of the NF-κB pathway and a marked decrease of the ERK signaling pathway in resistant cells. This was accompanied by lower glycolytic and glutaminolytic activities, possibly linked to glutamine being required to increase stemness capacity and, hence, higher survival to cisplatin. The TCA cycle dynamics seemed to be time-dependent, with an apparent activation at 48 h preferentially supported by anaplerotic aromatic amino acids, leucine and lysine. A distinct behavior of leucine, compared to the other branched-chain-amino-acids, suggested the importance of the recognized relationship between leucine and in mTOR-mediated autophagy to increase resistance. Suggested markers of MDA-MB-231 TNBC cisplatin-resistance included higher phosphocreatine/creatine ratios, hypotaurine/taurine–mediated antioxidant protective mechanisms, a generalized marked depletion in nucleotides/nucleosides, and a distinctive pattern of choline compounds. Although the putative hypotheses generated here require biological demonstration, they pave the way to the use of metabolites as markers of cisplatin-resistance in TNBC and as guidance to develop therapies. [ABSTRACT FROM AUTHOR]

Nanoparticles are widely used in the pharmaceutical, agriculture, and food processing industries. In this study, we have synthesized green lead nanoparticles (gPbNPs) by using an extract of Ziziphus spina-christi leaves and determined their cytotoxic and apoptotic effect on the human breast cancer MDA-MB-231 cell line. gPbNPs were characterized by using X-ray diffraction (XRD), energy dispersive X-ray (EDX) scanning electron microscope (SEM), and transmission electron microscope (TEM). The toxicity of gPbNPs was determined on the MDA-MB-231 cell line using MTT and NRU assays and as a result cell viability was reduced in a concentration-dependent manner. MDA-MB-231 cells were more sensitive at the highest concentration of gPbNPs exposure. In this experiment, we observed the production of intracellular ROS in cells, and induction of caspase 3/7 was higher in cells at 42 µg/ml of gPbNPs. Moreover, the Bax gene was upregulated and the Bcl-2 gene was downregulated and increased caspase 3/7 activity confirmed the apoptotic effect of gPbNPs in cells. Our observation showed that gPbNPs induced cell toxicity, increased generation of intracellular ROS, and gene expression of Bcl-2 and Bax in the MDA-MB-231 cell line. In conclusion, these findings demonstrated that gPbNPs executed toxic effects on the MDA-MB-231 cell line through activating caspase 3/7 activity. [ABSTRACT FROM AUTHOR]

Low-temperature vacuum drying (LTVD) has shown great potential for drying vegetables. It could avoid excessive degradations of active compounds with potential therapeutic agents. In this study, the effect on several relevant bioactive compounds, anti-inflammatory activity, and anti-proliferative activity of broccoli (Brassica oleracea var. italica) were evaluated. Effects of other drying methods, including vacuum drying (VD), convective drying (CD), infrared drying (IRD), and freeze drying (FD), were also comparatively evaluated. The results of all dried samples showed high polyunsaturated fatty acid contents (of up to 71.3%) and essential amino acid contents (of up to 8.63%). The LTVD method stands out above the other drying methods, since it obtained the highest content of total phenols, chlorogenic acid, and ferulic acid. Both the LTVD and CD samples demonstrated high anti-inflammatory and anti-proliferative activities. These CD and LTVD samples were also the most active against the breast carcinoma MDA-MB-23 cell line. Due to the good retention of bioactive compounds via LTVD, the obtained dried broccoli here can be used in a near time as an ingredient for the development of novel natural products with anti-inflammatory and anti-proliferative effects. [ABSTRACT FROM AUTHOR]

Purpose: Although chemotherapy and radiotherapy in conjunction with surgery have been known as the standard methods for patients with breast cancer, they frequently face resistance due to the failure of cells to death. Accordingly, improving the results requires discovering novel therapeutic approaches based on the changes in the molecular biology of cancer cells. Osteopontin (OPN) is a secreted protein that previous studies have shown to be associated with progression, poor prognosis, and metastasis in breast cancer. The current study examined the synergistic effects of radiotherapy and knocking out of OPN gene, utilizing CRISPR/Cas9 technique in MDA-MB-231 breast cancer cells. Methods: We used to knock out the OPN gene by the two different gRNAs. The cells irradiated 24 h after transfection. The mRNA expression, tumor cell proliferation, cell cycle distribution, growth, and apoptosis were measured. Moreover, activation of Chk1 and AKT were measured via western blot. Results: We demonstrated the OPN knocking out along with radiation led to the promotion of apoptosis, suppression of downstream genes, reduction of cell viability, and inhibition of cell-cycle progression. The western blot analysis has indicated that the knocking out of the OPN gene along with radiotherapy changes DNA damage responses substantially. Conclusions: The OPN gene knocking out with radiotherapy might be an efficient approach to overcome the radioresistance in breast cancer. [ABSTRACT FROM AUTHOR]

Simple Summary: Animal testing is an important method in medical research and the development of new drugs (pharmaceuticals). Classic animal models feature mice or rats, but whenever possible, these animals should only be used for scientific purposes when absolutely necessary. Using less-developed forms, such as embryos not capable of pain perception, is considered an approach to reduce and replace adult animal testing. The use of chicken embryos for cancer research is well-known and features the implantation of cancer cells in order to form tumors (xenografts) on embryonic membranes (CAM). The tumor can be investigated via imaging methods, for example, nuclear medical imaging. However, chicken embryos are small and, therefore, require dedicated small animal imaging systems, which are expensive and require trained personnel. Therefore, we have investigated whether large ostrich embryos also are capable of growing tumors on the CAM. The large size of ostrich embryos would allow the use of routine imaging devices used for examinations in humans. We implanted breast cancer cells on the ostrich embryo CAM and successfully observed tumor growth. We suggest that the ostrich embryo is a suitable model for xenograft tumor imaging and cancer-related pharmaceutical research. This needs to be elucidated in further studies. The avian chorioallantoic membrane (CAM) assay has attracted scientific attention in cancer research as an alternative or complementary method for in vivo animal models. Here, we present a xenograft model based on the ostrich (struthio camelus) CAM assay for the first time. The engraftment of 2 × 106 breast cancer carcinoma MDA-MB-231 cells successfully lead to tumor formation. Tumor growth monitoring was evaluated in eight fertilized eggs after xenotransplantation. Cancer cells were injected directly onto the CAM surface, close to a well-vascularized area. Histological analysis confirmed the epithelial origin of tumors. The CAM of ostrich embryos provides a large experimental surface for the xenograft, while the comparably long developmental period allows for a long experimental window for tumor growth and treatment. These advantages could make the ostrich CAM assay an attractive alternative to the well-established chick embryo model. Additionally, the large size of ostrich embryos compared to mice and rats could help overcome the limitations of small animal models. The suggested ostrich model is promising for future applications, for example, in radiopharmaceutical research, the size of the embryonal organs may compensate for the loss in image resolution caused by physical limitations in small animal positron emission tomography (PET) imaging. [ABSTRACT FROM AUTHOR]

The purpose of this study was to develop a carbohydrate silica nanocomposite, M-mDOX@SiO2, that can specifically target cancer cells and release the drug doxorubicin (DOX) specifically in an acidic tumor microenvironment (TME). First, DOX@SiO2 was prepared using the Stöber method to encapsulate DOX within silica nanoparticles (NPs). DOX@SiO2 was then immersed in acidic PBS (pH 5) to obtain mesoporous silica-based NPs, termed mDOX@SiO2. Through mannoside and galactoside surface silane modification of mDOX@SiO2, we obtained M-mDOX@SiO2 (specifically targeting the MDA-MB-231 cell surface receptor) and G-mDOX@SiO2 (a nontargeting control), respectively. The synthesized silica NPs were characterized through ultraviolet–visible (UV–vis) spectrophotometry, fluorescence spectrometry, dynamic light scattering, and transmission electron microscopy. The anthrone–sulfuric acid method was used to measure the NP surface sugar content, and thermogravimetric analysis was applied to estimate the content of DOX loaded inside the NPs. The drug release profiles of M-mDOX@SiO2 in different pH buffer solutions demonstrated that M-mDOX@SiO2 can effectively release DOX in an environment with a pH of 5; moreover, the drug release rate increased as pH decreased. Fluorescence spectrometry and confocal microscopy were used to image MDA-MB-231 cells specifically targeted by M-mDOX@SiO2. The MTT assay results revealed that M-mDOX@SiO2 resulted in efficient cell growth inhibition. Finally, our animal experiments revealed that M-mDOX@SiO2 demonstrated high antitumor efficacy. In conclusion, M-mDOX@SiO2 has great potential as a drug delivery carrier. [ABSTRACT FROM AUTHOR]

Aberrant composition of glycans in the tumor microenvironment (TME) contributes to tumor progression and metastasis. Chondroitin polymerizing factor (CHPF) is a glycosyltransferase that catalyzes the biosynthesis of chondroitin sulfate (CS). It is also correlated to transforming growth factor-β1 (TGF-β1) expression, a crucial mediator in the interaction of cancer cells with TME. In this study, we investigated the association of CHPF expression with the clinicopathological features of breast cancer (BRCA), as well the oncogenic effect and the underling mechanisms of CHPF upon BRCA cells. We found that CHPF expression is significantly increased in human BRCA tissues, and it is positively associated with TGF-β expression (r = 0.7125). The high-expression of CHPF predicts a poor prognosis and is positively correlated with tumor mass, lymph node metastasis, clinical staging and HER-2 negative-expression. The mechanistic study revealed that it promotes BRCA cell proliferation, migration and invasion through TGF-β1-induced SMAD3 and JNK activation in vitro, JNK (SP600125) or SMAD3 (SIS3) inhibitor can remove the promotion of CHPF upon cell proliferation, migration and invasion in MDA-MB-231 cells, which is derived from triple-negative breast cancer (TNBC). Collectively, our finding suggested CHPF may function as an oncogene and is highly expressed in human BRCA tissues. Pharmacological blockade of the upstream of JNK or SMAD3 signaling may provide a novel therapeutic target for refractory TNBC patients with CHPF abnormal high-expression. [ABSTRACT FROM AUTHOR]

The growth response and biological activity of the cyanobacterium Oscillatoria sancta were investigated in starvation conditions. Oscillatoria sancta growth potential was examined on BG11 and Zarrouk's media. Zarrouk's medium supported the maximum growth of the test cyanobacterium. Zarrouk's medium composition was modified by excluding CaCl2·2H2O, NaCl, EDTA (Na), micronutrients, and replacing sodium nitrate with urea. Using Zarrouk's medium and three different concentrations of modified Zarrouk's media (Treatments 1–3), the growth response of Oscillatoria sancta (MZ366482) was examined and compared. Zarrouk's medium and modified Zarrouk's medium at 12.5% nutrient concentration had non-significant differences in both the dry weight biomass and total protein of Oscillatoria sancta. Oscillatoria sancta crude biomass extracts grown on Zarrouk's and modified Zarrouk's media (T3) inhibited human breast cancer, pathogenic bacteria, and acetylcholinesterase activity. Oscillatoria sancta grown on T3 showed the most potency against MDA-MB-231 cells with an IC50 of 165.2 µg mL−1, antibacterial activity only against Bacillus cereus (17.2 mm) and Staphylococcus aureus (15.3 mm), and acetylcholinesterase inhibition activity by 60.7%. Thus, it is advisable to use the 12.5% nutrient concentration of modified Zarrouk's medium as a reduced-cost medium for mass cultivation of Oscillatoria sancta with potential anticancer, antibacterial diseases, and anti-Alzheimer purposes. [ABSTRACT FROM AUTHOR]

Research background. Breast cancer is one of the most common cancers and remains a major cause of morbidity and mortality among women worldwide. In developed countries, breast cancer as a multifactorial disease is a major health concern, and its incidence is constantly rising in low and middle-income countries. Numerous studies have demonstrated that phytochemicals such as carotenoids inhibit breast cancer growth and induce apoptosis. We recently enhanced the solubility of capsanthin in water by encapsulating it in diosgenin polyethylene glycol succinate, a novel non-ionic surfactant. Thus, this study aims to evaluate the cytotoxicity of water-soluble capsanthin-loaded micelles in MDA-MB-231 cells in vitro through tetrazolium dye MTT assay. Experimental approach. In the current study, capsanthin, a hydrophobic carotenoid, is extracted from sweet red pepper (Capsicum annuum). Capsanthin-loaded diosgenin polyethylene glycol succinate 1000 (cap-DPGS-1000) micelles were prepared from capsanthin extract (cap) and diosgenin polyethylene glycol succinate 1000 (DPGS-1000) using the solid dispersion method. The capsanthin extract and cap-DPGS-1000 micelles were characterized by UV-visible spectroscopy, high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), particle size distribution, polydispersity, and scanning electron microscopy (SEM). The effects of capsanthin extract and cap-DPGS-1000 micelles on a human triple-negative breast cancer cell line (MDA-MB-231) were tested to check the cell viability, proliferation and cytotoxicity of the micelles. Results and conclusions. The solubility of encapsulated cap-DPGS-1000 micelles in water is greatly enhanced and leads to an increased scope for localized drug delivery, a better delivery option for treating residual cancerous tumours. The encapsulated capsanthin showed a sustained release in simulated intestinal fluid (pH=6.8). Our research proposes a sustained drug delivery system that ensures effective and controlled release to the affected site. The characterization data revealed no change in the structure and functional groups in the encapsulated capsanthin. The IC50 value of the cap-DPGS-1000 micelles against MDA-MB-231 breast cancer cells was (3.10±1.09) μg/mL, which is much lower than of capsanthin extract ((81.1±1.5) μg/mL). Capsanthin extract and capsanthin-loaded micelles are promising drug candidates to induce apoptosis and increase reactive oxygen species (ROS) in cancer cells. Novelty and scientific contribution. The result shows the cytotoxic effect of capsanthin and capsanthin-loaded micelles on MDA-MB-231 cell line for the first time. Capsanthin from sweet red pepper (Capsicum annuum) showed remarkable cytotoxic effect on the triple-negative MDA-MB-231 cell line.

Studies have shown that simulated microgravity (SMG) affects tumor cell growth and metastasis. However, the underlying molecular basis is still not known. In recent years, due to the high expression of CD44 in invasive basal breast tumors, it has been the subject of many studies. The aim of present study was to investigate the gene expression of CD44 in MDA-MB-231cell line of breast cancer in microgravity conditioncell line was proliferated under normal gravity and microgravity (1 and 3 days) using 2-D clinostat. Gene expression was measured using real-time PCR technique. SMG increased gene expression (100%) after 1 day and decreased it (15%) during 3 days in comparison to the control samples. It seems that the response of cancer cells to microgravity is time dependent and simulated microgravity treatment for 3 days may have a positive effect on cancer characteristics of MDA-MB-231 cell line in order to decrease the expression of CD44.

Low-temperature vacuum drying (LTVD) has shown great potential for drying vegetables. It could avoid excessive degradations of active compounds with potential therapeutic agents. In this study, the effect on several relevant bioactive compounds, anti-inflammatory activity, and anti-proliferative activity of broccoli (Brassica oleracea var. italica) were evaluated. Effects of other drying methods, including vacuum drying (VD), convective drying (CD), infrared drying (IRD), and freeze drying (FD), were also comparatively evaluated. The results of all dried samples showed high polyunsaturated fatty acid contents (of up to 71.3%) and essential amino acid contents (of up to 8.63%). The LTVD method stands out above the other drying methods, since it obtained the highest content of total phenols, chlorogenic acid, and ferulic acid. Both the LTVD and CD samples demonstrated high anti-inflammatory and anti-proliferative activities. These CD and LTVD samples were also the most active against the breast carcinoma MDA-MB-23 cell line. Due to the good retention of bioactive compounds via LTVD, the obtained dried broccoli here can be used in a near time as an ingredient for the development of novel natural products with anti-inflammatory and anti-proliferative effects.

The avian chorioallantoic membrane (CAM) assay has attracted scientific attention in cancer research as an alternative or complementary method for in vivo animal models. Here, we present a xenograft model based on the ostrich (struthio camelus) CAM assay for the first time. The engraftment of 2 × 106 breast cancer carcinoma MDA-MB-231 cells successfully lead to tumor formation. Tumor growth monitoring was evaluated in eight fertilized eggs after xenotransplantation. Cancer cells were injected directly onto the CAM surface, close to a well-vascularized area. Histological analysis confirmed the epithelial origin of tumors. The CAM of ostrich embryos provides a large experimental surface for the xenograft, while the comparably long developmental period allows for a long experimental window for tumor growth and treatment. These advantages could make the ostrich CAM assay an attractive alternative to the well-established chick embryo model. Additionally, the large size of ostrich embryos compared to mice and rats could help overcome the limitations of small animal models. The suggested ostrich model is promising for future applications, for example, in radiopharmaceutical research, the size of the embryonal organs may compensate for the loss in image resolution caused by physical limitations in small animal positron emission tomography (PET) imaging.

Background: The assembly and disassembly of the focal adhesions (FA) components occurs throughout life cycle of adhesion, with conservation of balance between removal and recruitment rate during temporal stages. Previous studies have demonstrated that phosphotidyilinositols play a role in regulating FA turnover. However, a little attention has been given to quantify the dynamics changes of Phosphatidylinositol 3,4,5- trisphosphate (PtdIns (3,4,5) P3) within and during fast and slow turnover rates of FA. Methods: MDA-MB-231 breast cancer cell line was used as a model in this study due to high metastatic and motile. These cells were co-transfected with GFP- paxillin/vinculin, as FA marker, and the GFP/mCherry-Btk-PH, as a biosensor to visualize PtdIns (3,4,5) P3. Confocal time-lapse images were used to monitor changes or differences in the local generation of PtdIns (3,4,5) P3 within and during assembly and disassembly of FA. Following transfection, immunostaining was used to examine the spatial colocalization between FA and PtdIns (3,4,5) P3. Results: Our data demonstrated that PtdIns (3,4,5) P3 co-localized with FAs and increase during assembly and decline during disassembly of FA which exhibits slow turnover rates and was in a constant level during assembly and disassembly of FA that displays fast turnover rates. Conclusions: Our result suggested that the dynamic changes of PtdIns (3,4,5) P3, it may depend on components undergo turnover, such that early, nascent FA displays fast turnover rates and mature FA exhibits slow turnover rates. Thus, the local enrichment of PtdIns (3,4,5) P3 enhances FA assembly and disassembly activation. [ABSTRACT FROM AUTHOR]

Research background. Breast cancer is one of the most common cancers and remains a major cause of morbidity and mortality among women worldwide. In developed countries, breast cancer as a multifactorial disease is a major health concern, and its incidence is constantly rising in low and middle-income countries. Numerous studies have demonstrated that phytochemicals such as carotenoids inhibit breast cancer growth and induce apoptosis. We recently enhanced the solubility of capsanthin in water by encapsulating it in diosgenin polyethylene glycol succinate, a novel non-ionic surfactant. Thus, this study aims to evaluate the cytotoxicity of water-soluble capsanthin-loaded micelles in MDA--MB-231 cells in vitro through tetrazolium dye MTT assay. Experimental approach. In the current study, capsanthin, a hydrophobic carotenoid, is extracted from sweet red pepper (Capsicum annuum). Capsanthin-loaded diosgenin polyethylene glycol succinate 1000 (cap-DPGS-1000) micelles were prepared from capsanthin extract (cap) and diosgenin polyethylene glycol succinate 1000 (DPGS-1000) using the solid dispersion method. The capsanthin extract and cap-DPGS-1000 micelles were characterized by UV-visible spectroscopy, high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), particle size distribution, polydispersity, and scanning electron microscopy (SEM). The effects of capsanthin extract and cap-DPGS-1000 micelles on a human triple-negative breast cancer cell line (MDA-MB-231) were tested to check the cell viability, proliferation and cytotoxicity of the micelles. Results and conclusions. The solubility of encapsulated cap-DPGS-1000 micelles in water is greatly enhanced and leads to an increased scope for localized drug delivery, a better delivery option for treating residual cancerous tumours. The encapsulated capsanthin showed a sustained release in simulated intestinal fluid (pH=6.8). Our research proposes a sustained drug delivery system that ensures effective and controlled release to the affected site. The characterization data revealed no change in the structure and functional groups in the encapsulated capsanthin. The IC50 value of the cap-DPGS-1000 micelles against MDA-MB-231 breast cancer cells was (3.10±1.09) μg/mL, which is much lower than of capsanthin extract ((81.1±1.5) μg/mL). Capsanthin extract and capsanthin-loaded micelles are promising drug candidates to induce apoptosis and increase reactive oxygen species (ROS) in cancer cells. Novelty and scientific contribution. The result shows the cytotoxic effect of capsanthin and capsanthin-loaded micelles on MDA-MB-231 cell line for the first time. Capsanthin from sweet red pepper (Capsicum annuum) showed remarkable cytotoxic effect on the triple-negative MDA-MB-231 cell line. [ABSTRACT FROM AUTHOR]

The endoplasmic reticulum (ER) chaperone Grp94/gp96 appears to be involved in cytoprotection without being required for cell survival. This study compared the effects of Grp94 protein levels on Ca2+ homeostasis, antioxidant cytoprotection and protein–protein interactions between two widely studied cell lines, the myogenic C2C12 and the epithelial HeLa, and two breast cancer cell lines, MDA-MB-231 and HS578T. In myogenic cells, but not in HeLa, Grp94 overexpression exerted cytoprotection by reducing ER Ca2+ storage, due to an inhibitory effect on SERCA2. In C2C12 cells, but not in HeLa, Grp94 co-immunoprecipitated with non-client proteins, such as nNOS, SERCA2 and PMCA, which co-fractionated by sucrose gradient centrifugation in a distinct, medium density, ER vesicular compartment. Active nNOS was also required for Grp94-induced cytoprotection, since its inhibition by L-NNA disrupted the co-immunoprecipitation and co-fractionation of Grp94 with nNOS and SERCA2, and increased apoptosis. Comparably, only the breast cancer cell line MDA-MB-231, which showed Grp94 co-immunoprecipitation with nNOS, SERCA2 and PMCA, increased oxidant-induced apoptosis after nNOS inhibition or Grp94 silencing. These results identify the Grp94-driven multiprotein complex, including active nNOS as mechanistically involved in antioxidant cytoprotection by means of nNOS activity and improved Ca2+ homeostasis. [ABSTRACT FROM AUTHOR]

Breast cancer (BC) is the most prevalent cancer and the one with the highest mortality among women worldwide. Although the molecular classification of BC has been a helpful tool for diagnosing and predicting the treatment of BC, developments are still being made to improve the diagnosis and find new therapeutic targets. Mitochondrial dysfunction is a crucial feature of cancer, which can be associated with cancer aggressiveness. Although the importance of mitochondrial dynamics in cancer is well recognized, its involvement in the mitochondrial function and bioenergetics context in BC molecular subtypes has been scantly explored. In this study, we combined mitochondrial function and bioenergetics experiments in MCF7 and MDA-MB-231 cell lines with statistical and bioinformatics analyses of the mitochondrial proteome of luminal A and basal-like tumors. We demonstrate that basal-like tumors exhibit a vicious cycle between mitochondrial fusion and fission; impaired but not completely inactive mitochondrial function; and the Warburg effect, associated with decreased oxidative phosphorylation (OXPHOS) complexes I and III. Together with the results obtained in the cell lines and the mitochondrial proteome analysis, two mitochondrial signatures were proposed: one signature reflecting alterations in mitochondrial functions and a second signature exclusively of OXPHOS, which allow us to distinguish between luminal A and basal-like tumors. [ABSTRACT FROM AUTHOR]

Introduction: Gravity is one of the most important forces exposed to the organisms on the earth which affects the cells, molecules and thus the entire of organism. The purpose of this study is to investigate the effect of the simulated microgravity on the changes of RKIP metastasis suppressor gene expression on MDA-MB-231 breast cancer cell line.Materials and Methods: In order to impose microgravity state, cancer cells were placed on clinostat for one and three days. Then RNA was extracted from the cells and RKIP gene expression changes were evaluated by qReal time PCR.Results: The findings showed that the microgravity during one day reduced the RKIP expression level, but with continuing up to three days, the expression returned to the control level.Conclusion: By optimizing the duration of microgravity, it can be likely observed the significant effects on the RKIP gene expression changes and so that useful steps can be taken to discover the cancer mechanisms and its treatment.

This study aimed to investigate the effect of Catharanthus roseus L. (C. roseus) leaf extract on the migration and invasion of MDA-MB-231 cell line and elucidate the molecular mechanisms of action. Effect of the extract on cell viability was evaluated by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl2H-tetrazoliumbromide) assay. Anti-migratory and anti-invasive effects were evaluated using scratch and Transwell assays. Effect on the levels and activities of matrix metalloproteinase (MMP)-2 and MMP-9 was determined using ELISA and gelatin zymography. Furthermore, changes in the expression of 84 genes commonly involved in cell motility were assessed by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and cell motility RT2 profiler PCR array. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using DAVID. Using STRING and Cytoscape software, hub genes were determined. The extract significantly (p < 0.001) inhibited the migration and invasion of MDA-MB-231 cells at non-cytotoxic concentrations. The activities and levels of MMP-2 and MMP-9 were decreased in a dose-dependent manner following C. roseus exposure. At 4 µg/mL, the extract significantly downregulated the expression of 52 genes involved in extracellular matrix degradation, cytoskeleton reorganisation, focal adhesions and invadopodia formation. GO and KEGG pathway analysis revealed that the downregulated genes were significantly enriched in biological processes and pathways closely related to cell motility. Our findings showed that C. roseus inhibited the migration and invasion of MDA-MB-231 cells through altering the expression of various motility-related genes. This study provided data about the potential of C. roseus phytochemicals as promising therapeutic agents against breast cancer metastasis, especially at gene level. [ABSTRACT FROM AUTHOR]

Objectives: To screen the cytotoxic activity of six secondary metabolites isolated from soil fungus Aspergillus niger. Importantly, to investigate the mechanism that pyoluteorin induced human triple‐negative breast cancer MDA‐MB‐231 cells apoptosis in vitro. Methods: The cell viability assay was tested with CTG assay. Cell cycle, apoptosis and intracellular reactive oxygen species (ROS) production assay were tested with flow cytometry. Additionally, intracellular ROS production assay and mitochondrial membrane potential assay were determined with laser scanning confocal microscopy. The expression of apoptosis‐related proteins was determined with Western blot. Key findings: Pyoluteorin displayed significantly selective cytotoxicity against human triple‐negative breast cancer MDA‐MB‐231 cells (IC50 = 0.97 µm) with low toxicity against human breast epithelial cell MCF‐10A. It was found that pyoluteorin could arrest MDA‐MB‐231 cells cycle at G2/M phase and induce cell apoptosis. Further experiments demonstrated that the apoptosis‐inducing effect of pyoluteorin was related to reduction of mitochondrial membrane potential, accumulation of ROS and change of apoptosis‐related protein expressions. Conclusion: Our studies revealed that pyoluteorin had potent proliferation inhibition against MDA‐MB‐231 cells through arresting cell cycle at G2/M phase and inducing caspase‐3‐dependent apoptosis by mitochondrial pathway, implying that pyoluteorin may be a potential lead compound for drug discovery of human triple‐negative breast cancer. [ABSTRACT FROM AUTHOR]

[Display omitted] • Synthesis and characterization of fluorescence receptors N1, N2 -bis(2-piperazine-1-yl) ethyl) oxalamide [OX(PIP) 2 ] and Di-(benzoyloxalohydrazide) [OX(BH) 2 ] for detecting Sn2+ and Fe3+ ions. • Absorbance and fluorescence spectral activity of the OX(PIP) 2 & OX(BH) 2 receptors for Sn2+/Fe3+ ions. • Gelation ability of receptor with various solvent. • CT-DNA binding studies with fluorescence receptors with their Sn2+/Fe3+ ions. • Investigate the antibacterial activity, anticancer activity and cell imaging properties. The development of colorimetric and fluorescence receptors N1, N2 -bis(2-piperazine-1-yl)ethyl)oxalamide [OX(PIP) 2 ] and Di-(benzoyloxalohydrazide) [OX(BH) 2 ] for the accurate and easily identified Sn2+ & Fe3+ ions respectively in DMSO/HEPES buffer (1:9, v/v, 20 μM, pH 7.3) was a success. The process of sensing entails the complexation of OX(PIP) 2 receptors with Sn2+ addition, which sets off the fluorescence 'Turn-On' mode and OX(BH) 2 displays 'Turn-Off' mode after the inclusion of Fe3+ ions. An improved linear correlation between fluorescence intensity and Sn2+/Fe3+ concentration was attained in the range of 0–20 μM, with a detection limit (LOD) of 0.6 μM & 0.013 μM respectively. The average fluorescence lifespan measurements of OX(PIP) 2 and OX(BH) 2 were determined to be 2.77 and 2.22 ns, and the average life time of 4.49 ns for OX(PIP) 2 + Sn2+ and 2.18 ns for OX(BH) 2 + Fe3+ were observed. The binding mechanisms of OX(PIP) 2 + Sn2+ and OX(BH) 2 + Fe3+ were confirmed by Fourier transform infrared analysis, NMR spectral titrations, and mass (ESI) spectral analysis. The chemosensing of OX(PIP) 2 and OX(BH) 2 has also been examined in bioimaging experiments as useful fluorescent markers for finding the source of Sn2+/Fe3+ in living cells. Significantly, receptors OX(PIP) 2 and OX(BH) 2 could distinguish Sn2+/Fe3+ ions in live cells and Sn2+/Fe3+ in human cancer cells. [ABSTRACT FROM AUTHOR]

Perfluoroalkyl substances (PFASs) are a category of high-concerned emerging contaminants which are suspected to correlate with various human adverse health outcomes including tumors. It is also a question whether short-chain PFASs are qualified alternatives under the regulation of long-chain PFASs. In this study, a three-dimensional (3D) culture system based on Gelatin methacrylate (GelMA) hydrogel matrix was used to investigate the impacts of 120-h perfluorooctanoic acid (PFOA) and perfluorobutanoic acid (PFBA) exposure of MDA-MB-231 cells. The results showed that PFOA exposure promoted the proliferation, migration, and invasion of MDA-MB-231 cells in an environmentally relevant concentration range (0.1 to 10 μM), exhibiting a clear malignant-promoting risk. In contrast, PFBA only showed a trend to induce non-invasive cell migration. Hippo/YAP signaling pathway was identified as the contributor to the differences between the two PFASs. PFOA but PFBA reduced YAP phosphorylation and increased the nuclear content of YAP, which further facilitated abundant key factors of epithelial-mesenchymal transition (EMT) process. Our results provided a new idea for the carcinogenicity of PFOA using a 3D-based paradigm. Although the effects by PFBA were much milder than PFOA in the current test duration, the cell model suitable for longer exposure is still necessary to better assess the safety of alternative short-chain PFASs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Abstract Background The expression of p38 MAPK is high in breast cancer while its subunit p38γ had been rarely reported. We aimed to explain the effect of p38γ in breast cancer from the perspective of metabolomics. Methods In this study, we detected the expression of p38γ in 28 breast carcinoma and para-tumor samples. Following MDA-MB-231 cell transfection with p38γ siRNAs and pc-DNA-3.1, cell viability, apoptosis, metastasis were determined through CCK-8, the cytometry analysis, transwell assay and wound healing assay. Finally, gas chromatograph-mass spectrometer (GC–MS) was used for analysis the differential metabolites. Results The expression of p38γ was significantly up-regulated in breast cancer tissues. The transfection of si-p38γs could inhibit MDA-MB-231 cell propagation, metastasis, and induced cell apoptosis while overexpressed p38γ could promote the cell propagation, metastasis, and inhibit cell apoptosis. A total of 238 metabolites were identified and 72 of them differentially expressed in three groups (all P < 0.05, FDR < 0.05). Then the metabolites were enriched in the metabolism pathway, 85 pathways were included and 27 were significant (all P < 0.05, FDR < 0.05). Conclusions p38γ was up-regulated in breast cancer, which exerts a great influence on the cell growth, cell mobility, invasiveness, and apoptosis of MDA-MB-231 cells and also affected the metabolism. [ABSTRACT FROM AUTHOR]

Propolis has been used in folk medicine for centuries due to its healing properties. Ethanolic extracts of propolis (EEP) are rich sources of phenolic acid and flavonoids. Natural phenolic compounds may exert chemoprotective activity in cancer cells due to their ability to scavenge free radicals. The aim of this in vitro study was to investigate the genotoxic and anti-mutagenic effects of the EEP on human peripheral blood lymphocytes (PBLs) and their cytotoxic potential on the human breast cancer cell line (MDA-MB-231 cells). Both cell cultures were treated with six concentrations (1, 10, 50, 100, 250 and 500 μg/ml) of EEP1 and EEP2, separately and in combination with mitomycin C (MMC). Our results show that the EEP1 and EEP2 samples of propolis after separate and combined treatments with MMC did not influence the nuclear division index (NDI). In the combined treatment, both tested EEPs significantly reduced MMC-induced micronuclei (MN) in PBLs. At 48 h after exposure of the MDA-MB-231 cell line to a combined treatment of EEP samples with MMC, the IC50 values were significantly reduced (23.79 and 19.13 μg/ml, for EEP1+MMC and EEP2+MMC, respectively, in comparison to the single treatment. In conclusion, the tested ethanolic extracts of propolis exhibited a certain level of in vitro antimutagenic activity in PBLs from healthy subjects, and anticancer activity in breast cancer cell line. The presented findings suggest that the ethanolic extracts of propolis show potential in anticancer therapeutic strategy. [Projekat Ministarstva nauke Republike Srbije, br. III41010]

• ZnPc-PDT or TA were cytotoxic to triple-negative breast cancer cells, MDA-MB-231 cell line. • The combination of ZnPc-PDT and TA was more cytotoxic compared with their alone on the MDA-MB-231 cells. • ZnPc-PDT could enhance the sensitivity of the MDA-MB-231 cells towards TA treatment. Photodynamic therapy (PDT) is a light-based anti-neoplastic therapeutic approach. Growing evidence indicates that combining conventional anti-cancer therapies with PDT can be a promising approach to treat malignancies. Herein, we aimed to investigate anti-cancer effects of the combination treatment of zinc phthalocyanine (ZnPc)-PDT with tamoxifen (TA) on MDA-MB-231 cells (as a triple-negative breast cancer (TNBC) cell line). For this purpose, we investigated the cytotoxicity of TA and ZnPc-PDT on MDA-MB-231 cells performing the MTT assay. The effect of TA and ZnPc-PDT on the apoptosis of MDA-MB-231 cells was studied using Annexin V/PI and DAPI staining. The wound-healing assay, and colony formation assay were performed to study the effect of TA and ZnPc-PDT on the migration, and clonogenicity of MDA-MB-231 cells, respectively. The qRT-PCR was done to study the gene expression of caspase-8, caspase-9, caspase-3, ZEB1, ROCK1, SNAIL1, CD133, CD44, SOX2, and ABCG2 (ATP-binding cassette sub-family G member 2). Based on our results, monotherapies with TA and ZnPc-PDT can remarkably increase cell cytotoxicity effects, stimulate apoptosis via downregulating Bcl-2 and upregulating caspase-3 and caspase-9, inhibit migration via downregulating SNAIL1 and ZEB1, and suppress clonogenicity via downregulating SOX2 and CD44 in MDA-MB-231 cells. Besides, these monotherapies can downregulate the expression of ABCG2 in MDA-MB-231 cells. Nevertheless, the combination treatment can potentiate the above-mentioned anti-cancer effects compared to monotherapy with TA. Of interest, the combined treatment of TA with ZnPc-PDT can synergically increase cell cytotoxicity effects on MDA-MB-231 cells. In fact, synergistic effects were estimated by calculation of Combination Index (CI); that synergistic outcomes were observed in all groups. Also, this combination treatment can significantly upregulate the caspase-8 gene expression and downregulate ROCK1 and CD133 gene expression in MDA-MB-231 cells. Overall, our results show that ZnPc-PDT can more sensitize the MDA-MB-231 cells to TA treatment. Based on our knowledge and experiment, the synergistic effects of ZnPc-PDT and TA deserve further evaluation in cancer research. [ABSTRACT FROM AUTHOR]

Background: Due to the aggressive clinical behavior, poor outcome, and lack of effective specific targeted therapies, triple-negative breast cancer (TNBC) has currently been recognized as one of the most malignant types of tumors. In the present study, we investigated the cytotoxic effect of ziyuglycoside I, one of the major components extracted from Chinese anti-tumor herbal Radix Sanguisorbae, on the TNBC cell line MDA-MB-231. Methods: The underlying molecular mechanism of the cytotoxic effect ziyuglycoside I on MDA-MB-231 cells was investigated with cell viability assay, flow cytometric analysis andWestern blot. Results: Compared to normal mammary gland Hs 578Bst cells, treatment of ziyuglycoside I resulted in a significant growth inhibitory effect on MDA-MB-231 cells. Ziyuglycoside I induced the G2/M phase arrest and apoptosis of MDA-MB-231 cells in a dose-dependent manner. These effects were found to be partially mediated through the up-regulation of p53 and p21WAF1, elevated Bax/Bcl-2 ratio, and the activation of both intrinsic (mitochondrial-initiated) and extrinsic (Fas/FasL-initiated) apoptotic pathways. Furthermore, the p53 specific siRNA attenuated these effects. Conclusion: Our study suggested that ziyuglycoside I-triggered MDA-MB-231 cell cycle arrest and apoptosis were probably mediated by p53. This suggests that ziyuglycoside I might be a potential drug candidate for treating TNBC. [ABSTRACT FROM AUTHOR]

Drug resistance in breast cancer remains a major obstacle of clinical therapy. We found that suppression of ELK3 in the triple negative breast cancer cell line MDA-MB-231 impaired autophagy and led to a hypersensitive response to doxorubicin treatment. In ELK3-knockdown MDA-MB-231 cells (ELK3 KD), autophagy was not activated under starvation conditions, which is a major stimulus of autophagy activation. We revealed that activation of the PI3K/Akt pathway was the main cause of impaired autophagy in ELK3 KD. Our results suggest that targeting ELK3 may be a potential approach to overcome doxorubicin resistance in breast cancer therapeutics. [ABSTRACT FROM AUTHOR]

Effective control of breast cancer has been primarily hampered by a lack of tumor specificity in treatments. One potential way to improve targeting specificity is to develop novel vectors that specifically bind to and are internalized by tumor cells. Through a phage display library, an 11-L-amino acid peptide, PI (sequence, CASPSGALRSC), was selected. PI was labeled with fluorescein isothiocyanate (FITC) and named PI-FITC. Subsequently, the specific affinity of PI-FITC to MDA-MB-231 human breast cancer cells and other cancer cell lines was observed by confocal microscopy. Our previous study established that PI-FITC also shows affinity to Calu-1 human lung carcinoma cells and major histocompatibility complex class I antigen molecules; therefore, the cytomembrane proteins of the cell lines were analyzed to determine those that were common to the two cell lines and may be associated with transmembrane transduction. To further test the delivery ability of PI to MDA-MB-231 cells, PI-glutathione-S-transferase (GST) was constructed and the internalization of this fusion protein was visualized by immunofluorescence microscopy. The results revealed that PI exhibited specific affinity to MDA-MB-231 cells. Use of membrane transport inhibitors indicated that macropinocytosis and caveolin-mediated endocytosis may be involved in the endocytosis of PI. In addition, 11 membrane proteins common to MDA-MB-231 and Calu-1 may be associated with transmembrane transduction. In summary, PI was able to deliver PI-GST into MDA-MB-231 cells. Thus, PI could be modified to be a potential vector, and may contribute to the development of targeted therapeutic strategies for breast cancer. [ABSTRACT FROM AUTHOR]

Rab5A and Akt pathways are reported to be responsible for the invasiveness of cancer cells, indicated by the fact that Rab5A activates the downstream Phosphoinositide-3-kinases (PI3K)/Akt signalling pathway, which results in promoting cancer metastasis. However, little attention has been given to the emerging role of Rab5A and Akt signalling pathways in regulating the direction of MDA-MB-231 cell migration. MDA-MB-231 breast cancer cell line was used as a model in this study because it is highly metastatic and motile. Time-lapse microscopy was used to examine the effect of Akt and Rab5A inhibitors on cell migration, proliferation and wound healing. Later, the cells were transfected with GFP-Akt-PH or GFP-Rab5A (used as a biosensor to detect Akt and Rab5A). Therefore, confocal time-lapse images were used to visualize Akt and Rab5A at the front and rear edges of the cells. The recorded data demonstrated that Akt and Rab5A inhibition reduced cell migration, proliferation and wound healing. The results of the current study also demonstrated that Akt localizes at the trailing edge while Rab5A localize more at the leading edge than the trailing edge of cells. This study suggests that Akt and Rab5A inhibition might regulate the direction of breast cancer migration., Competing Interests: The authors declare that they have no conflict of interest.

The efforts described in this article are aimed at designing organotin polymers that control the growth of breast cancer and to identify structure/property relationships that assist in this goal. The growth of MCF-7 and MDA-MB-231 breast cancer cell lines is inhibited employing a wide range of organotin condensation polymers. The EC50values are primarily dependent on the nature of the Lewis base but the CI50is dependent on both the nature of the Lewis base and Lewis acid. A number of products exhibit CI50values greater than two including a number of organotin polyethers such as those derived from diethylstilbestrol, dienestrol, short-chained dibutyltin polyethers, and hydroquinone derivatives. In most of these cases the MDA-MB-231 cells exhibit greater inhibition compared to the estrogen receptor (ER) MCF-7 cells. The organotin polymers generally exhibit a superior ability to inhibit MCF-7 and MDA-MB-231 breast cell growth compared to the standard cisplatin. [ABSTRACT FROM PUBLISHER]

Cysteine-rich protein 61(CCN1/Cyr61) has been implicated as an important mediator in proliferation and metastasis of breast cancer, which indicated that blockage of Cyr61 might be a potent target for breast cancer treatment. However, the antitumor effect of anti-Cyr61 antibodies on breast cancer in vivo has not been reported so far. In this study, we reported the effect and likely mechanism of generated anti-human Cyr61 monoclonal antibodies (mAb) on Cyr61 high expression line MDA-MB-231, known as a highly malignant and invasive human breast cancer cell line, at aspects of proliferation and migration in vitro and in vivo. We found the mAb, denoted as 093G9, revealed inhibitory effects on MDA-MB-231 cell proliferation, migration, and invasion through downregulation of both AKT and ERK phosphorylation in vitro compared with its isotype control. 093G9 also showed significant efficacy on suppressing primary tumor growth and spontaneous lymph node metastasis in in vivo mouse model. The specific epitope recognized by 093G9 was identified to be LPNLGCP, adjacent to the VWC domain of Cyr61 by Ph.D.-C7C phage library display system. Our study provides direct evidence that Cyr61 can be a potent therapeutic target for patients who bear high Cyr61 expression breast cancer. Furthermore, the mAb, 093G9 developed in our laboratory, has shown a promising therapeutic characteristic in breast cancer. [ABSTRACT FROM AUTHOR]