Treatment Of Breast Cancer Cells Research Articles

Network pharmacology and experimental validation to reveal the pharmacological mechanisms of Qizhu prescription for treating breast cancer

ObjectiveTo investigate the mechanism underlying the effects exerted by the Qizhu prescription (QZP) in breast cancer (BC), and the respective targets. MethodsExpression data from the ArrayExpress and The Cancer Genome Atlas (TCGA) were used to identify differentially expressed genes (DEGs) in BC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the DEGs to identify genes involved in protein–protein interactions. Molecular docking was used to explore the dynamic relationship between active molecules and targets. Cell function experiments and animal studies were conducted to evaluate the effects of hub genes and active QZP compounds on BC cell behavior. ResultsAmong the 25 evaluated BC-related targets of QZP, matrix metalloproteinase-1 (MMP1) and epidermal growth factor receptor (EGFR) exhibited the highest degrees of dysregulation. GO and KEGG enrichment analyses revealed that the anti-BC targets of QZP primarily affected drug responses and pathways in cancer cells. Molecular docking analysis suggested potential interactions between EGFR and quercetin/luteolin, as well as between MMP1 and luteolin/kaempferol/quercetin. Quercetin significantly reduced BC cell proliferation, migration, invasion, and tumor development in vivo. Treatment of BC cells with quercetin decreased the expression or activation of several associated proteins. ConclusionThe findings of our study provide new insights into the therapeutic potential of traditional Chinese medicine against BC, with particular reference to QZP.

Exploring the Anticancer Activity of Gold Complex with Newly Ligand (DDIBM): Synthesis, Spectral Identification and Magnetic Susceptibility of Its Metallic Complexes

The new heterocyclic ligand, 5-(dimethylamino)-2-(((2-((E)-(4,5-diphenyl-1H-imidazol-2-yl)diazenyl)benzyl)imino)methyl)phenol (DDIBM), was synthesized via the condensation of p-aminobenzylamine with 4,5-diphenyl imidazole, and the resultant compound was condensed with 4-(dimethylamino)-2-hydroxybenzaldehyde. Various instrumental techniques such as mass, 1H-NMR, IR, C.H.N elemental analysis, and UV-vis spectroscopy were used to analyze a newly synthesized ligand. A novel series of complexes was prepared by complexing the ligand with Ni(II), Cu(II), Co(II), and Au(III) and characterized using some of the mentioned techniques. Flame atomic absorption spectroscopy was used to measure the metal ion percentages in the complexes. The magnetic susceptibility and molar conductivity were studied. The electronic spectral data and the magnetic measurement predict the octahedral structure of the complexes except Au(III) complex which has square planer geometry. All complexes showed electrolyte properties. This study aimed to conduct an in vitro cytotoxicity comparative study of DDIBM and its Au(III) complex on human breast cancer cells (MCF-7) and other normal cells. The Au(III) complex was found to be highly selective in targeting cancer cells without affecting normal healthy cells, compared to the ligand. Thus, this complex can be considered as a new drug for treating breast cancer cells (MCF-7), and an attempt in the future to study its effect on other types of cancer.

Molecular insights into quercetin-enhanced crystalline silver nanoparticles: Implications for breast cancer cell cytotoxicity and antibacterial action in Kyllinga nemoralis

Amidst the ongoing global health crisis posed by cancer and antibiotic-resistant infections, innovative solutions are in high demand. The utilization of Kyllinga nemoralis leaf (KNL) extracts for synthesizing silver nanoparticles (AgNPs) represents a novel approach, with the potential to deliver profound advancements in healthcare, benefitting society on a broad scale. Successful synthesis of AgNPs by KNL extracts for treating breast cancer cells, specifically MDA-MB-231 cells, is recorded in this report. The adsorption mechanism of quercetin (Q) on Ag and its optimized structural characteristics were computed at the DFT level. According to FTIR analysis, quercetin, one of the flavonoids found in KNL extract, acts as a reducing and stabilizing agent for the conversion of Ag+ ions to Ag0, and carboxylates serve as ligands that cap the AgNPs. The small-sized (7 nm), spherical particles, which are uniform in size and monodispersed, exhibit regulation over the size of the AgNPs due to being shielded by enough flavonoids and gallic acid absorbed into the extract. The observed negative zeta potential of −20 mV indicates the moderate stability of the AgNPs colloid. It is noted that AgNPs prepared at higher concentrations of KNL extract showed lower minimum inhibitory concentration (MIC) values (80 µg/mL) and larger inhibition zones (22 mm) against Pseudomonas aeruginosa. The cell viability rate of MDA-MB-231 cell lines decreases with increasing concentration of AgNPs. The antibacterial and anticancer activity of AgNPs is observed in a size-dependent manner. This research raises the possibility of using plant-based, small-sized AgNPs therapeutically for breast cancer and as antibacterial agents.

Abstract 1222: Mechanisms of succinate-mediated drug resistance in ER+ breast cancer

Abstract Breast cancer is the most prevalent cancer among women worldwide with estrogen receptor-positive (ER+) cancer constituting 70% of all breast cancers. Although ER+ breast cancers initially respond well to ER-targeted therapies and CDK4/6 inhibitors, approximately 30% patients acquire resistance to targeted therapies. In addition to intrinsic resistance that tumors develop, cancer-associated fibroblasts (CAFs) residing in the tumor microenvironment, also contribute to treatment-resistance and poor survival. The role of secreted metabolites by CAFs has been poorly studied in the context of targeted drug resistance. Therefore, to investigate the mechanism of stroma-mediated resistance to CDK4/6-inhibitors, we propagated patient-derived organoids (PDOs) and CAFs from treatment-naïve and treatment-resistant ER+ breast tumors. Our data show that the CAF conditioned media containing the polar metabolite fraction stimulates resistance to CDK4/6 inhibitors. Therefore, to identify metabolites that drive resistance, we performed an untargeted metabolomics on CAF conditioned media. We observed that the CAF conditioned media was enriched for TCA cycle intermediates, including succinate. Succinate is an oncometabolite that has an essential function in energy metabolism. Aside from causing metabolic reprogramming, succinate can be transported out of the mitochondria to perform other biological processes, such as epigenetic modifications and pseudohypoxia. Furthermore, succinate in the extracellular space binds to the succinate receptor SUCNR1 and induces G-protein coupled receptor-mediated signaling in cells expressing SUCNR1. To assess mechanism of resistance that these metabolites might induce, we treated breast cancer cells with the most enriched metabolites we had identified in our metabolomics assay. These experiments identified succinate as driver of CDK4/6 inhibitor resistance in ER+ breast cancer cells. Furthermore, treating tumor cells with SUCNR1 antagonist increased their sensitivity to CDK4/6-inhibitors, showing that succinate mediated resistance is conveyed via SUCNR1. This suggests that CAF secreted succinate plays a role in supporting a pro-tumor environment by promoting drug resistance towards CDK4/6 inhibitors. Defining this mechanism of resistance will elucidate potential therapeutic targets to augment patient response to therapy. Citation Format: Jenny Hogstrom, Boryana Petrova, Camila Perea, Madelyn Ward, Gerburg Wulf, Laura Collins, Naama Kanarek, Taru Muranen. Mechanisms of succinate-mediated drug resistance in ER+ breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1222.

Abstract 607: Development of a novel, selective, and potent inhibitor of RON kinase

Abstract Purpose: Receptor d’origine nantais (RON) is a highly conserved transmembrane protein expressed at low levels in healthy adult tissues of epithelial origin and at various levels in immune cells such as macrophages. Aberrant activation of RON has been described in many solid tumors, and it contributes to tumorigenesis by modulating the immune tumor microenvironment, activating numerous oncogenic signaling pathways, and protecting tumor cells under stress. The overexpression of RON protein and the subsequent generation of oncogenic variants are mainly responsible for the persistent activation of downstream signaling pathways. In breast cancer, more than 80% of primary cancer samples are positive for RON expression, with overexpression reported in approximately 50% of cases. Importantly, tumoral RON expression correlates with increased breast cancer progression, metastasis, and poor prognosis independent of molecular subtype. We hypothesize that a small molecule inhibitor of RON that can target both full-length and truncated isoforms will promote anti-tumor immunity within the tumor microenvironment and block pro-metastatic signaling pathways. Herein, we report the development of a novel, orally bioavailable, selective, and potent inhibitor of RON kinase. Methods: With structure-guided and iterative medicinal chemistry approaches, we identified a novel chemical entity that potently inhibited RON kinase activity. Further optimization resulted in the identification of ZB-60 with improved potency, selectivity, and desirable drug-like properties. In vitro kinase assays were performed using ADP-Glo RON kinase assay kit (Promega). Changes in phosphorylation of RON was demonstrated by Western blotting. Plasma pharmacokinetics (PK) was conducted in mice to measure exposures and to determine the oral bioavailability of ZB-60. In vivo efficacy of ZB-60 was evaluated in syngeneic mouse models of breast (EMT6) and colorectal (CT26) cancer. Results: ZB-60 inhibited RON kinase activity with an IC50 of 56 nM and showed greater than 40-fold selectivity against MET and other closest members of the tyrosine kinase family. In ZB-60 treated breast cancer cells, there was a dose dependent reduction of RON phosphorylation and downstream signaling. PK analysis showed a high oral bioavailability with low clearance. In vivo evaluation of ZB-60 showed inhibition of tumor growth in EMT6 and CT26 models. Ongoing studies are focused on testing ZB-60 in combination with checkpoint inhibitors. The results from these studies along with ADME-TOX and pharmacokinetics will be presented. Conclusion: ZB-60 has target specific activity and exhibits favorable drug-like properties. Citation Format: Mohan Rao Kaadige, Trason Thode, Jaime Fornetti, Alexis Weston, Serina Ng, Tithi Ghosh-Halder, Taylor Bargenquast, Brian Durbin, Srinivas Kasibhatla, Raffaella Soldi, Alana Welm, Sunil Sharma. Development of a novel, selective, and potent inhibitor of RON kinase [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 607.

MiR-218-5p and doxorubicin combination enhances anticancer activity in breast cancer cells through Parkin-dependent mitophagy inhibition

Breast Cancer (BC) is one of the most common tumours, and is known for its ability to develop resistance to chemotherapeutic treatments. Autophagy has been linked to chemotherapeutic response in several types of cancer, highlighting its contribution to this process. However, the role of mitophagy, a selective form of autophagy responsible for damaged mitochondria degradation, in the response to therapies in BC is still unclear. In order to address this point, we analysed the role of mitophagy in the treatment of the most common anticancer drug, doxorubicin (DXR), in different models of BC, such as a luminal A subtype-BC cell line MCF7 cells, cultured in 2-Dimension (2D) or in 3-Dimension (3D), and the triple negative BC (TNBC) cell line MDA-MB-231. Through a microarray analysis, we identified a relationship between mitophagy gene expressions related to the canonical PINK1/Parkin-mediated pathway and DXR treatment in BC cells. Afterwards, we demonstrated that the PINK1/Parkin-dependent mitophagy is indeed induced following DXR treatment and that exogenous expression of a small non-coding RNA, the miRNA-218-5p, known to target mRNA of Parkin, was sufficient to inhibit the DXR-mediated mitophagy in MCF7 and in MDA-MB-231 cells, thereby increasing their sensitivity to DXR. Considering the current challenges involved in BC refractory to treatment, our work could provide a promising approach to prevent tumour resistance and recurrence, potentially leading to the development of an innovative approach to combine mitophagy inhibition and chemotherapy.

Exploring MTH1 inhibitory potential of Thymoquinone and Baicalin for therapeutic targeting of breast cancer

Cancers frequently have increased ROS levels due to disrupted redox balance, leading to oxidative DNA and protein damage, mutations, and apoptosis. The MTH1 protein plays a crucial role by sanitizing the oxidized dNTP pools. Hence, cancer cells rely on MTH1 to prevent the integration of oxidized dNTPs into DNA, preventing DNA damage and allowing cancer cell proliferation. We have discovered Thymoquinone (TQ) and Baicalin (BC) as inhibitors of MTH1 using combined docking and MD simulation approaches complemented by experimental validations via assessing binding affinity and enzyme inhibition. Docking and MD simulations studies revealed an efficient binding of TQ and BC to the active site pocket of the MTH1, and the resultant complexes are appreciably stable. Fluorescence measurements estimated a strong binding affinity of TQ and BC with Ka 3.4 ×106 and 1.0 ×105, respectively. Treating breast cancer cells with TQ and BC significantly inhibited the growth and proliferation (IC50 values 28.3 µM and 34.8 µM) and induced apoptosis. TQ and BC increased the ROS production in MCF7 cells, imposing substantial oxidative stress on cancer cells and leading to cell death. Finally, TQ and BC are proven strong MTH1 inhibitors, offering promising prospects for anti-cancer therapy.

Extracellular vesicles derived from oxidatively stressed stromal cells promote cancer progression

Like many diseases, cancer is influenced by oxidative stress. Increases in oxidative stress caused by factors such as environmental stressors and unhealthy diets can increase the risk of cancer and worsen cancer prognoses. However, oxidative stress does not always promote cancer. On the contrary, many chemotherapeutics induce the production of oxidative stress to kill cancer cells. The tumor microenvironment (TME) is another factor in cancer progression. Stromal cells, a part of the TME, communicate with cancer cells through extracellular vesicles (EVs) - the primary vehicles for the delivery of messengers, including proteins, metabolites, DNA, and RNA. We hypothesized that the TME mediates cancer’s response to oxidative stress by delivering EVs to cancer cells. In order to test this hypothesis, we treated breast cancer cells (HCC1806, SKBR3, MCF7, and MDA-MB-436) and lung cancer cells (A549 and PC9) with EVs derived from oxidatively stressed stromal cells and compared their expression of marker proteins of cell proliferation and epithelial to mesenchymal transition to cancer cells treated with EVs from control stromal cells and untreated cancer cells. We found that EVs extracted from oxidatively stressed adipocytes increased the cell proliferation of breast cancer cells. Additionally, we found that EVs extracted from lung fibroblasts promoted epithelialmesenchymal transition in lung cancer cells. These findings present a novel way that the TME influences cancer progression. The findings of this paper can inform future cancer prevention studies.

Formononetin Restrains Tumorigenesis of Breast Tumor by Restraining STING-NF-κB and Interfering with the Activation of PD-L1.

Breast cancer (BC), a common tumor in women, has high morbidity and mortality. Formononetin, an active ingredient in red clover and Astragalus membranaceus, has a wide range of pharmacological applications, including as an anticancer agent. Since immunotherapy is a hot topic in the treatment strategy of BC, it was dedicated to appraising the specific mechanism of formononetin in BC immunotherapy in this research. Different formononetin concentrations (0, 20, 40, 60, 80, 100 μM) were used to treat BC cells transfected with pcDNA3.1-Programmed death ligand 1 (PD-L1) or Short-hairpin RNA (sh)-PD-L1. Cells were separated into four subgroups: CTRL, pcDNA3.1-PD-L1, sh-CTRL, and sh-PD-L1. Cell viability and cell cycle were assessed through Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and flow cytometry. Programmed death ligand 1 (PD-L1) mRNA concentration was validated via quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Cell metastasis was evaluated via cloning assay and transwell assay. The p-STING/stimulator of interferon genes (STING), p-p65/p65, and PD-L1 concentrations were determined by western blot. Formononetin restrained the proliferation of MCF-7 and MDA-MB-468 cells, and reduced PD-L1 mRNA, p-STING/STING, and p-p65/p65 protein concentrations. Whereas PD-L1 inhibition restrained the viability of BC cells, pcDNA3.1-PD-L1 intervention had the opposite result. STING pathway inhibitor C-176 combined with formononetin treatment further restrained cell proliferation, colony formation, and cell invasion, in contrast to cells treated with formononetin alone. Formononetin can restrain the proliferation of BC cells, which may be mediated through the interference of PD-L1 and suppression of the activation of the STING-NF-κB pathway.

MUC1 aptamer-conjugated niclosamide-loaded PLGA-PEG nanoparticles attenuate HIF-1 stabilization upon hypoxia in MCF7 breast cancer cells

Hypoxia is related to aggressive phenotype and poor prognosis in breast cancer. HIF-1, a transcription factor (TF), plays an essential role in the adaptation of tumor cells to hypoxia. It is stabilized in hypoxic conditions and activates other TFs and signaling pathways that are associated with the proliferation, survival, and progression of cancer cells in hypoxia. As a result, targeting hypoxia by destabilizing HIF-1 is a promising approach to inhibit cell adaptation to hypoxia. Niclosamide (NIC) has been identified as a HIF-1 inhibitor and it is envisioned that the anti-cancer efficacy of NIC can be improved through targeted drug delivery systems (DDSs) as a therapeutic strategy. Here, MUC1 aptamer (Ap)-targeted PLGA/PEGylated nanoparticles (Ap-PLGA-NPs) were developed for targeted delivery of NIC (Ap-NIC-PLGA-NPs) to MUC1 positive breast cancer cell line in hypoxia environment. The obtained findings showed that targeted inhibition of HIF-1 using fabricated targeted-NIC-loaded NPs increased the rate of apoptosis in breast cancer cells in hypoxic conditions which was further confirmed by geno-/cytotoxicity assays and flow cytometry analysis. Downregulation of HIF-1α, HIF-1β, Eno1, and GluT1 in treated breast cancer cells under hypoxic conditions also indicated a potential role for Ap-NIC-PLGA-NPs in tumor cell adaptation to hypoxia.

Anti-breast cancer activity of biosynthesized selenium nanoparticles using Bacillus coagulans supernatant

BackgroundIn the present study, Selenium Nanoparticles (SeNPs) were prepared using Bacillus coagulans, which is a type of Lactic Acid Bacteria (LAB), and then they were applied to treat breast cancer cells. MethodsThe chemicophysical properties of the bioengineered SeNPs were investigated by Transmission Electron Microscopy (TEM), Field Emission Scanning Electron Microscopy (FE-SEM), zeta potential, dynamic light scattering, Fourier Transform Infrared Spectroscopy (FT-IR), energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction analysis (XRD). The cytotoxic potential of SeNPs was evaluated by MTT assay against MCF-7 breast cancer cell line. The expression levels of apoptotic genes including BAX, BCL2, VEGF, ERBB2, CASP3, CASP9, CCNE1, CCND1, MMP2 and MMP9 were determined by real-time PCR. The rate of apoptosis and necrosis of the cancer cells as well as the results of the cell cycle were evaluated by flow cytometry method. ResultsThe synthesized SeNPs had an average particle size of about 24–40 nm and a zeta potential of –16.1 mV, indicating the high stability of SeNPs. EDX results showed presence of SeNPs because amount of selenium in SeNPs was 86.6 % by weight. The cytotoxicity results showed a concentration-dependent effect against MCF-7 cells. The half-maximal inhibitory concentration (IC50) values of B. coagulans supernatant and SeNPs against breast cancer cells were 389.7 µg/mL and 17.56 µg/mL, respectively. In addition, SeNPs synthesized by the green process exhibited enhanced apoptotic potential in MCF-7 cancer cells compared with bacterial supernatants. Cancer cells treated with IC50 concentration of SeNPs induced 32 % apoptosis compared to untreated cells (3 % apoptosis). The gene expression levels of BAX, CASP3, and CASP9 were upregulated, while the expression levels of BCL2, CCNE1, CCND1, MMP2, MMP9, VEGF, and ERBB2 were downregulated after SeNPs treatment of cells. The potential of SeNPs to induce cell apoptosis was demonstrated by the increase in the expression level of BAX gene and the decrease in the expression level of BCL2 after treatment of cancer cells with SeNPs. ConclusionThe obtained results indicated that SeNPs had strong potential to induce significant cell apoptosis and are cytotoxic against the MCF-7 cancer cell line.

β-eudesmol inhibits cell proliferation and induces ferroptosis via regulating MAPK signaling pathway in breast cancer

The aim of this study was to explore the influences and underlying mechanisms of β-eudesmol on breast cancer (BC). Different concentrations of β-eudesmol (0, 10, 20, and 40 μM) were taken to treat BC cells. Cell Counting Kit-8, colony formation assay, and flow cytometry were performed to evaluate the influences of β-eudesmol on cell viability, proliferation, and apoptosis. To assess the influences of β-eudesmol on cell ferroptosis, the change of ROS, SOD, MDA, and intracellular iron and Fe2+ were determined. The protein changes of apoptosis, ferroptosis, and MAPK pathway (Bcl-2, Bax, cleaved caspase-3, SLC7A11, GPX4, SLC40A1, Transferrin, MEK1, and ERK1/2) were checked utilizing Western blot. In a concentration-dependent manner, β-eudesmol restrained cell viability and proliferation. β-eudesmol promoted cell apoptosis, as evidenced by the decline level of Bcl-2 and the raised level of Bax and cleaved caspase-3. β-eudesmol enhanced the level of ROS, MDA, iron, Fe2+, and Transferrin, and lessened SOD activity and the protein expression of SLC7A11, GPX4, SLC40A1, MEK1, and ERK1/2. Moreover, ferroptosis inhibitor Fer-1 and MEK1 overexpression both reversed the changes on cell proliferation, apoptosis, and ferroptosis induced by β-eudesmol. β-eudesmol inhibited cell proliferation and promoted cell apoptosis and ferroptosis via regulating MAPK pathway in BC.

Adverse effect of Tamarindus indica and tamoxifen combination on redox balance and genotoxicity of breast cancer cell

BackgroundBreast cancer is the most significant threat to women worldwide. Most chemotherapeutic drugs cause cancer cell death and apoptosis by inducing oxidative stress and producing reactive oxygen species (ROS). Cancer cells have a higher rate of metabolic activity than normal cells and thus produce more ROS. Glutathione and its related enzymes are the most significant antioxidant defense mechanisms that protect cells from oxidative and chemotherapeutic impacts. The anticancer actions of phenolic compounds were greatly confirmed. Using phenolic compounds as drugs in combination with chemotherapy may improve health, improve treatment outcomes, and reduce dose and damage. The goal of the study was to treat breast cancer cell lines (MCF-7) with Tamarindus indica extract individually and in combination with the anticancer drug tamoxifen (TAM) to improve therapeutic efficacy. ResultsAfter 48 h of incubation at IC25 concentrations of T. indica extract (47.3 g/mL), tamoxifen (0.8 g/mL), and their co-treatments, the biochemical and genotoxic effects on MCF-7 cell lines were investigated. In MCF7 cell lines, T. indica extract increased reduced glutathione levels as well as glutathione transferase, glutathione peroxidase, and glutathione reductase activities. The same was true for oxidative state indicators, where higher levels of catalase and lactate dehydrogenase activity were associated with higher levels of malondialdehyde. T. indica has almost no effect on the DNA damage parameters. All of these variations can produce alterations in cancer cell genotoxicity and apoptotic pathways, explaining the restoration of DNA moment to normal levels and enhanced survival. ConclusionCytotoxic and genotoxic effect of treatment with T. indica extract could be attributed to the dynamic interaction of glutathione cycle and antioxidant enzymes to combat oxidative stress, which can be considered as a positive therapeutic effect. On the other hand, the negative response of tamoxifen efficacy when co-treated with T. indica reversed tamoxifen’s genotoxicity and enhanced survival.

Efficacy of copper nanoparticles encapsulated in soya lecithin liposomes in treating breast cancer cells (MCF-7) in vitro

Cancer is one of the leading causes of death, which has attracted the attention of the scientific world to the search for efficient methods for treatment. With the great development and regeneration of nanotechnology over the last 25 years, various nanoparticles in different structures, shapes and composites provide good potential for cancer therapy. There are several drugs approved by FDA used in breast cancer treatment like Cyclophosphamide, Doxorubicin Hydrochloride, Femara, Herceptin, etc. Each has several side effects as well as treatment, which limits the use of drugs due to heart failure, pulmonary dysfunction, or immunodeficiency. Recently, such side effects are greatly reduced by using innovative delivery techniques. Some drugs have been approved for use in cancer treatment under the concept of drug delivery, such as Doxil (liposomal loaded doxorubicin). The purpose of this study is to investigate the effect of copper nanoparticles (CuNPs) as a drug model for cancer treatment, either in their free form or encapsulated in Soy lecithin liposomes (SLP) from plant origin as a cheap source of lipids. CuNPs were prepared by the chemical reduction method and loaded onto SLP through the thin film hydration method. The drug model Cu/SLP was successfully combined. The characteristics of the free CuNPs, liposomes, and the combined form, zeta potential, size distribution, drug encapsulation efficiency (EE%), drug release profile, Fourier transform infrared (FTIR), and transmission electron microscopy (TEM), were checked, followed by an in vitro study on the breast cancer cell line Mcf-7 as a model for cytotoxicity evaluation. The optimal Cu/SLP had a particle mean size of 81.59 ± 14.93 nm, a negative zeta potential of − 50.7 ± 4.34 mV, loaded CuNPs showed an EE% of 78.9%, a drug release profile for about 50% of the drug was released after 6 h, and FTIR analysis was recorded. The cytotoxicity assay showed that the IC50 of Cu/SLP is smaller than that of free CuNPs. These results give clear evidence of the efficacy of using the combined Cu/SLP rather than CuNPs alone as a model drug carrier prepared from plant origin against cancer, both medically and economically.

Apoptosis Induction by ZnFe2O4-Ag Biosynthesized by Chlorella vulgaris in MCF-7 Breast Cancer Cell Line.

The incidence and mortality of breast cancer are growing which indicates the inefficiency of the current chemotherapy drugs. Due to the anticancer potential of Zn and Ag and the magnetic feature of iron oxide, in this work, we synthesized ZnFe2O4-Ag nanocomposite using Chlorella vulgaris and investigated its anticancer effect on breast cancer cell line. Physicochemical characterization was performed by FT-IR, XRD, SEM, TEM, VSM, EDS mapping, UV, and zeta potential assays. Cell cytotoxicity and apoptosis frequency were studied by the MTT and flow cytometry assays. Also, cell cycle analysis, Hoechst staining, and measuring ROS (reactive oxygen species) level were performed. The synthesized particles were almost spherical with a size range of 14-52 nm. The FT-IR and XRD assays confirmed the proper synthesis of the particles and VSM analysis showed that particles had magnetic property and the maximum saturation magnetization was 0.8 Emu/g. Also, the EDS mapping of the nanocomposite showed the Zn, Fe, O, and Ag elements. The MTT assay showed that the 50% inhibitory concentration (IC50) of ZnFe2O4-Ag for breast cancer and normal cells were 28 and 154µg/mL, respectively, and the nanocomposite had stronger anticancer activity than cisplatin (IC50 = 84µg/mL). Flow cytometry analysis showed that the exposure to the nanocomposite induced cell apoptosis by 77.5% and significantly induced ROS generation. Also, treating breast cancer cells with the nanocomposite induced cell cycle arrest and apoptotic features, including chromatin condensation and fragmentation. In conclusion, ZnFe2O4-Ag nanocomposite synthesized by C. vulgaris could suppress the proliferation of breast cancer cells by the generation of oxidative stress, apoptosis induction, and cell cycle arrest.

Evaluation of the cytotoxic effect of GaFe2O4@Ag magnetic nanocomposite biosynthesized by Spirulina platensis on breast cancer cell line and evaluation of BAX, Bcl-2, CASP9 and MMP1 genes expression

Treating breast cancer is hardly achieved and finding efficient anticancer nanocompounds has gained great attention. In this work, the cytotoxic effect of GaFe2O4@Ag nanocomposite biosynthesized by Spirulina platensis on breast cancer cell line, and expression of the BAX, Bcl-2, CASP9 and MMP1 (Matrix Metallopeptidase 1) genes were evaluated. Physicochemical features of the nanocomposite were determined using the FT-IR, XRD, SEM, TEM, EDX-mapping, VSM, Zeta potential, and DLS analyses. The cytotoxic effect of the nanocomposite for MCF-7 and HEK-293 cells was evaluated by the MTT assay. Flow cytometry analysis, Caspase-3 activation assay, and Hoechst staining were performed to evaluate the apoptosis induction potential of the nanocomposite. Further, the relative expression of the Bcl-2, BAX, and CASP9, and MMP1 genes was determined by quantitative PCR assay. The prepared nanocomposite was spherical with a size range of 35–60 nm. The hydrodynamic size and zeta potential of the nanocomposite were 328 nm and −31.8 mv, respectively. GaFe2O4@Ag nanocomposite had a higher cytotoxic effect on breast cancer cells than normal human cells with the IC50 of 18.6 and 220 µg/mL, respectively. Treating breast cancer cells with the nanocomposite induced apoptosis among 85.2% of cells, increased caspase-3 activity by 4.3 folds, and caused apoptotic nuclear changes. Also, GaFe2O4@Ag reduced the expression of the Bcl-2 and MMP1 by 1.3 and 0.6 folds and up-regulated the BAX and CASP9 genes by 2.7 and 2.65 folds, respectively. Our results revealed that GaFe2O4@Ag was highly cytotoxic for breast cancer cells via triggering apoptosis pathways and could be considered as a novel and efficient agent against breast cancer, after further in-vivo experiments.

Sevoflurane suppresses the malignant progression of breast cancer via the hsa_circ_0000129/miR-578/EPSTI1 axis.

Sevoflurane (Sev) is a commonly used volatile anesthetic that might suppress the process of breast cancer. Also, circular RNAs (circRNAs) have been reported to partake in the pathogenesis of breast cancer. Accordingly, this research was designed to investigate the mechanism of hsa_circ_0005962 on Sev-mediated breast cancer development. Sev was applied to treat breast cancer cells. Cell proliferation ability, migration, invasion, and apoptosis were detected using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), Transwell, and flow cytometry assay. Proliferating cell nuclear antigen (PCNA), Matrix metallopeptidase 9 (MMP9), B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax), and Epithelial stromal interaction 1 (EPSTI1) were assessed using western blot assay. circ_0000129, microRNA-578 (miR-578), and EPSTI1 levels were determined using real-time quantitative polymerase chain reaction (RT-qPCR). Using bioinformatics software (Circinteractome and Targetscan), the binding between miR-578 and circ_0000129 or EPSTI1 were predicted, and proved using dual-luciferase reporter and RNA pull-down assay. The biological roles of circ_0000129 and Sevoflurane on tumor growth were analyzed using a xenograft tumor model in vivo. Sevoflurane blocked tumor cell proliferation, migration, invasion, and promoted apoptosis. Circ_0000129 and EPSTI1 expression were increased, and miR-578 was decreased in breast cancer cells. Also, they presented an opposite trend in Sev-treated tumor cells. Circ_0000129 upregulation might abolish Sev-mediated tumor progression in vitro. Mechanically, circ_0000129 can affect EPSTI1 expression by sponging miR-578. Sev might inhibit tumor growth by regulating circ_0000129 in vivo. Circ_0000129 relieved Sev-triggered suppression impacts on breast cancer development partly via the miR-578/EPSTI1 axis, which provides a new mechanism for studying mediated therapy of breast cancer treatment.

Antibacterial potential and cytotoxic activity of iron oxide nanoparticles conjugated with thymol (Fe3O4@Glu-Thymol) on breast cancer cells and investigating the expression of BAX, CASP8, and BCL-2 genes.

The use of metal nanoparticles (NPs) conjugated with natural herbal molecules in biomedical applications has been growing. In this work, we synthesized Iron oxide NPs conjugated with thymol (Fe3O4@Glu-Thymol) and investigated their antibacterial and anticancer potentials. Physicochemical features of the NPs were studied by FT-IR, EDS-mapping, XRD, DLS, zeta potential, and electron microscopy. The antibacterial activity of the NPs against Pseudomonas aeruginosa and anticancer activity for breast cancer cells was investigated by broth microdilution and MTT and flow cytometry assays, respectively. The expression of apoptosis signaling genes in breast cancer cells that were treated with the NPs was studied by qPCR assay. The NPs were spherical, in a size range of 40-66nm, without impurities, and with zeta potential and hydrodynamic size of -23mV and 185nm, respectively. Moreover, the FT-IR and XRD assays confirmed the proper synthesis of Fe3O4 and conjugation with thymol. The minimum inhibitory concentration of the NPs for P. aeruginosa strains was 64-128µg/mL. Our results showed that Fe3O4@Glu-Thymol was considerably more toxic for breast cancer cells than normal human cells and the 50% inhibitory concentration were 90.4 and 322µg/mL, respectively. Upon treating breast cancer cells with the NPs the frequency of cell apoptosis increased by 18.9%. Also, the expression of the BAX and CASP8 genes in NPs treated cells significantly increased by 1.75 and 2.25 folds, respectively while the BCL-2 gene remained almost constant. This study reveals that Fe3O4@Glu-Thymol has considerable potential to be used in biomedical fields.

Effect of CeO2/spherical silica and halloysite nanotubes engineered for targeted drug delivery system to treat breast cancer cells

Cerium oxide nanoparticles (CeO2 NPs) and flavonoid curcumin that has been widely studied for treating diseases involving high reactive oxygen species (ROS). In nanotherapeutics, the particle size, shape, metal oxide dispersity and surface properties of nanocarriers are vital for drug delivery and therapeutic efficiency. Here, cisplatin release behavior on cerium impregnated two different shaped nanocarriers, CeO2/monodispersed spherical silica (Sil) and CeO2/halloysite (Hal) nanotube was studied for potential anti-cancer therapies. For comparison, CeO2 impregnated mesoporous silica MCM-41, SBA-16, Hydroxyapatite and clay were prepared. Subsequently, the nanocomposites were coated with curcumin (25% wt/wt), and cisplatin (Cp) functionalization (5% wt/wt). 5wt%CeO2/Hal/Cp and 5wt%CeO2/Sil/Cp samples were pegylated using lyophilization technique. Physico-chemical analyses revealed the nanosized distribution of CeO2 and functionalization of cisplatin and curcumin. Cp release was studied using automated Franz cell and dialysis membrane techniques. The different structured nanocarriers delivering mechanism was studied by determining the drug kinetic release using four different kinetic models (first order, second order, Higuchi and Korsmeyer-Peppas). In vitro cytotoxicity assay of nano formulations along with free cisplatin and curcumin (Cur) were tested against the breast cancer cell line (MCF-7) for multiple timepoints by MTT assay. The results reveled the efficacy of 5wt%CeO2/Sil/Cp/Cur nanoparticles in delivering cisplatin. On the other hand, 5wt%CeO2/Hal/Cur nanoparticles enhanced the uptake of curcumin in comparison to free curcumin. Overall, pegylated CeO2/Silica nano formulation demonstrated an effective carrier to cisplatin for potential treatment of breast cancer.

Overexpression of cannabinoid receptor 2 is associated with human breast cancer proliferation, apoptosis, chemosensitivity and prognosis via the PI3K/Akt/mTOR signaling pathway.

The cannabinoid receptor 2 (CB2) is mainly involved in the immune system. However, although CB2 has been reported to play an anti-tumor function in breast cancer (BC), its specific mechanism in BC remains unclear. We examined the expression and prognostic significance of CB2 in BC tissues by qPCR, second-generation sequencing, western blot, and immunohistochemistry. We assessed the impacts of overexpression and a specific agonist of CB2 on the growth, proliferation, apoptosis, and drug resistance of BC cells in vitro and in vivo using CCK-8, flow cytometry, TUNEL staining, immunofluorescence, tumor xenografts, western blot, and colony formation assays. CB2 expression was significantly lower in BC compared with paracancerous tissues. It was also highly expressed in benign tumors and ductal carcinoma in situ, and its expression was correlated with prognosis in BC patients. CB2 overexpression and treatment of BC cells with a CB2 agonist inhibited proliferation and promoted apoptosis, and these actions were achieved by suppressing the PI3K/Akt/mTOR signaling pathway. Moreover, CB2 expression was increased in MDA-MB-231 cell treated with cisplatin, doxorubicin, and docetaxel, and sensitivity to these anti-tumor drugs was increased in BC cells overexpressing CB2. These findings reveal that CB2 mediates BC via the PI3K/Akt/mTOR signaling pathway. CB2 could be a novel target for the diagnosis and treatment of BC.