MCF-7 Human Breast Cancer Research Articles

Synthesis and in vitro evaluation of antimycobacterial activity of two palladium(II) complexes based on 5-alkyl-1,3,4-oxadiazol-2(3H)-thione derivatives

In this work, two palladium(II) complexes, namely, [Pd(L1)2(phen)] 1 and [Pd(L2)2(phen)] 2 (L1 = 5-heptyl-1,3,4-oxadiazole-2-(3H)-thione, L2 = 5-nonyl-1,3,4-oxadiazole-2-(3H)-thione and phen = 1,10-phenanthroline), were prepared and charactherized by conventional techniques. In both complexes, the spectral data indicated that L1 and L2 are coordinated to the metal in a monodentate manner through the sulfur atom (S−), while a phenanthroline molecule acting as a bidentate ligand completes the coordination sphere. This proposal was supported by DFT calculation of structures and NMR spectra. Subsequently, the free ligands and their metal complexes were tested in vitro against the human prostate cell lines PNT-2 (non-tumorigenic), LNCaP (androgen-sensitive prostate cancer) and PC-3 (castration-resistant prostate cancer), as well as in two human breast cancer cell lines, MCF7 (luminal) and MDA-MB231 (triple-negative). Regarding cytotoxicity, all compounds were considered inactive, since the IC50 values found were higher than 80 µM. The complexes were also evaluated for activity against Mycobacterium tuberculosis, and 1 showed good activity (MIC = 8.94 µg/mL), while 2 exhibited moderate to poor activity (MIC > 25.0 µg/mL), revealing that an increase in the aliphatic chain makes it less active. A conformation sampling was conducted for the complexes 1 and 2 using molecular dynamics simulation. The analysis provided a clear picture of the steric and electrostatic flexibility of these compounds, which is an important feature for multi-target drugs.

Pyrimidines-Based Heterocyclic Compounds: Synthesis, Cytoxicity Evaluation and Molecular Docking.

A variety of structurally different pyrimidines were synthesized. Elemental analysis, FT-IR, 1H NMR, and 13C NMR spectroscopy were used to confirm the chemical structures of all prepared compounds. The synthesized pyrimidines were screened against the growth of five human cancer cell lines (prostate carcinoma PC3, liver carcinoma HepG-2, human colon cancer HCT-116, human breast cancer MCF-7, human lung cancer A-549), and normal human lung fibroblasts (MRC-5) using MTT assay. Most of the screened pyrimidines have anti-proliferative activity on the growth of the PC3 cell line. Compounds 3b and 3d were more potent than the reference vinblastine sulfate (~2 to 3 × fold) and they can be considered promising leads for treating prostate cancer disease. Moreover, the screened compounds 3b, 3f, 3g, 3h, and 5 were assessed according to the values of their selectivity index (SI) and were found to be more selective and safer than vinblastine sulfate. Furthermore, using in silico computational tools, the physicochemical properties of all pyrimidine ligands were assessed, and the synthesized compounds fall within the criteria of RO5, thus having the potential to be orally bioavailable.

Synthesis and Discovery of Ligustrazine-Heterocycle Derivatives as Antitumor Agents.

Ligustrazine (TMP) is a natural pyrazine alkaloid extracted from the roots of Ligusticum Chuanxiong Hort, which has the potential as an antitumor agent. A series of 33 ligustrazine–heterocycle (TMPH) derivatives were designed, synthesized, and investigated via antitumor screening assays, molecular docking analysis, and prediction of drug-like properties. TMP was attached to other heterocyclic derivatives by an 8–12 methylene alkyl chain as a linker to obtain 33 TMPH derivatives. The structures were confirmed by 1H-NMR, 13C-NMR, and high-resolution mass spectroscopy spectral (HR-MS) data. The antiproliferative activity against human breast cancer MCF-7, MDA-MB-231, mouse breast cancer 4T1, mouse fibroblast L929, and human umbilical vein endothelial HUVEC cell lines was evaluated by MTT assay. Compound 12–9 displayed significant inhibitory activity with IC50 values in the low micromolar range (0.84 ± 0.02 µM against the MDA-MB-231 cell line). The antitumor effects of compound 12–9 were further evaluated by plate cloning, Hoechst 33 342 staining, and annexin V-FITC/PI staining. The results indicated that compound 12–9 inhibited the proliferation and apoptosis of breast cancer cells. Furthermore, molecular docking of compound 12–9 into the active site of the Bcl-2, CASP-3, and PSMB5 target proteins was performed to explore the probable binding mode. The 33 newly synthesized compounds were predicted to have good drug-like properties in a theoretical study. Overall, these results indicated that compound 12–9 inhibited cell proliferation through PSMB5 and apoptosis through Bcl-2/CASP-3 apoptotic signaling pathways and had good drug-like properties. These results provided more information, and key precursor lead derivatives, in the search for effective bioactive components from Chinese natural medicines.

Anticancer activity of micropropagated Hedychium coronarium rhizome essential oil

Cancer is a life threatening disease than any other diseases which is leading to death throughout the world. The natural products should be explored as anticancer agents which are not expensive, easily available and most beneficial to humans. Today the current research is being done on secondary metabolites and bioactive compounds present in the medicinal plants alongwith their therapeutic properties. Zingiberaceae family from centuries is used as spices and bears vast pharmacological activities including Hedychium coronarium. Also another most important characteristic of this family is the aromatic essential oil present in its leaves and rhizomes. In the present study, Hedychium coronarium rhizome essential oils were treated against human breast cancer cell line or MCF7 and human cervical cancer cell line or HeLa by MTT assay. For validation of the cytotoxic activity, MCF7 and HeLa cell were used against various concentrations of aromatic rhizome essential oil respectively. This study revealed that bioactive compound present in rhizome oil was most significantly active when tested against MCF7 and HeLa cell line.

Ferula communis L. (Apiaceae) Root Acetone-Water Extract: Phytochemical Analysis, Cytotoxicity and In Vitro Evaluation of Estrogenic Properties.

Ferula communis L. (F. communis) belongs to the Apiaceae family and is a herbaceous plant with various pharmaceutical properties, due to the different contents of bioactive compounds extracted mainly from its roots, as well as its leaves and rhizome. To date, this plant extract has demonstrated estrogenic, anti-inflammatory, antiproliferative, cytotoxic, antimicrobial and anti-neoplastic properties. Its estrogenic activity is justified by the presence of ferutinin, an ester of a sesquiterpenic alcohol that acts as an agonist for estrogen receptors, with a chemical formula equal to C22H3O4. The component present in F. communis responsible for the toxicity of the plant is ferulenol, a prenylated coumarin with the chemical formula C24H30O3. This compound is capable of inducing mortality via its strong anti-coagulant properties, leading to a lethal hemorrhagic syndrome, ferulosis, in animals that feed on a chemotype of F. communis containing a high amount of ferulenol. The removal of the component ferulenol makes extracts of Ferula non-toxic. In fact, the remaining prenylated coumarins are not present in concentrations sufficient to induce toxicity. The intake of high concentrations of the extract of this plant leads a double dose-dependent effect that is typical of sesquiterpenes such as ferutinin. Here, we assessed the cytotoxicity and the estrogenic properties of the F. communis phytocomplex obtained through extraction using a mixture of acetone and water. Among the active constituents of F. communis, the identification of ferutinin and ferulenol was performed using HPLC. The effects of the extract were evaluated, following the removal of ferulenol, on three cell lines: human breast cancer MCF-7, human cervical cancer HeLa and human osteoblastic sarcoma Saos-2. The choice of these cell lines was justified by the need to mimic certain processes which may occur in vivo and which are estrogen-dependent. The obtained results demonstrated that F. communis extract, in addition to possessing an estrogenic-like property, showed a dose-dependent effect. Low concentrations (0.1–0.8 μM) demonstrated a hyperproliferative effect, whereas higher concentrations (1.6–50 μM) were toxic. Therefore, this extract could be an excellent candidate to make up for a reduction or lack of estrogen.

Curcumin Induces MCF-7 Cells Pyroptosis via Autophagy/CTSB/NLRP3/ Caspase-1/GSDMD Signaling Pathway in Vitro and Vivo

Background: Curcumin, as a lipid-lowering drug, has been reported to be effective in the treatment of breast cancer. However, the underlying molecular mechanisms have not been completely investigated. Methods: MTT assay was used to determine the effect of curcumin on survival rate of MCF-7 cells. The effects of curcumin on tumor growth were observed in animal models of breast cancer. The positive reactions of Caspase-1, IL-1β and IL-18 were detected by immunohistochemistry. LC3, p62, CTSB, ASC, Pro-Caspase-1, GSDMD, NLRP3, Caspase-1, GSDMD-N, IL-1β and IL-18 were determined by Western blot in vitro and vivo. ELISA determined the release of extracellular IL-1β and IL-18. LDH release was measured. The expression level of CTSB in cytoplasm were determined by immunofluorescence assay. Cell proliferation, cell migration and tube formation assays were used to determine the abilities of cells. In this study, NLRP3 inflammasome inhibitor MCC950, cathepsin B inhibitor CA-074 ME and autophagy inhibitor 3-MA were used to act on cells to investigate the role of NLRP3 inflammasome, cathepsin B and autophagy in curcumin-induced pyro ptosis of MCF-7 breast cancer cells. Results: In mouse model of breast cancer, we observed that curcumin treatment significantly induced cell autophagy and pyro ptosis. In human breast cancer MCF-7 cells, we found that curcumin induced pyrophoric cell death was dependent on the activation of NLRP3/Caspase-1/GSDMD signaling pathway, which was CTSB-dependent. In addition, curcumin-induced cell autophagy caused lysosomal rupture and CTSB release. Furthermore, NLRP3 inhibitor (MCC950) significantly suppressed curcumin-induced pyro ptosis, as well as CTSB inhibitor (CA074 Me) and autophagy inhibitor (3-MA). Besides, we also found that curcumin suppressed cell proliferation, cell migration and tube formation, which could be reversed by inhibitors. Conclusions: In summary, our results demonstrated that curcumin induced MCF-7 cell pyro ptosis by the activation of autophagy/CTSB/NLRP3/Caspase-1/GSDMD signaling pathway. These findings offer novel insights into the potential molecular mechanisms of curcumin in treatment of breast cancer

Synthesis, spectral, antioxidant, in vitro cytotoxicity activity and thermal analysis of Schiff base metal complexes with 2,2′-Bipyridine-4,4′-dicarboxylic acid as co-ligand

The Schiff base ligand (H2L1) [H2L1 = (E)-2-hydroxy-N'-((2-hydroxynaphthalen-1-yl)-methylene) benzo hydrazide] and its metal (II) complexes of molecular formula [Co(H2L1)(L2)], [Ni(H2L1)(L2)], [Cu(H2L1)(L2)], [Zn(H2L1)(L2)], [Cd(H2L1)(L2)], where, L2 = 2,2′-Bipyridine-4,4′-dicarboxylic acid as co ligand were synthesized and characterised using various analytical techniques. Based on spectral analysis, the complexes are found to be square pyramidal and also the coordinating sites of the complexes are azomethine nitrogen, two phenolic oxygen from Schiff base ligand (H2L1) and two nitrogen from 2,2′-Bipyridine-4,4′-dicarboxylic acid. The antimicrobial activity of the ligand and complexes was determined by testing them against bacterial pathogens and fungal strains. All the synthesized complexes were more active than the free chelate against the tested microorganisms. According to our findings, the antioxidant activity of complexes by the DPPH method is concentration-dependant and better than the free ligand H2L1. Compared to the therapeutically utilised Sorafenib (Nexavar), the metal complexes and its H2L1 ligand displayed stronger cytotoxicity against the human breast cancer MCF -7 cell lines. The [Co(H2L1)(L2)] complex showed promising anticancer activity with IC50 3.5 μg/mL. Cell death is due to apoptosis in MCF-7 cells and the formation of ROS by the Schiff base ligand H2L1 and its derived metal complexes.

Cytotoxic and Antibacterial Properties of Resveratrol Oligomers from the Stem Bark of Dryobalanops rappa

Background: Dryobalanops rappa is a plant species belonging to the family of Dipterocarpaceae. Nevertheless, the active compounds present in D. rappa have never been investigated. Objective: The aim of this research is to isolate and characterize compounds from Dryobalanops rappa and to study its bioactivity against human MCF-7 breast cancer and A549 lung cancer cell lines and several bacterial strains. Methods: The isolation step was carried out using a combination of chromatographic techniques. The structure of the isolated compounds was elucidated mainly using NMR spectroscopy. The cytotoxic activity of isolated compounds was determined with MTT assay, and the antimicrobial was screened using a modified resazurin microtiter-plate assay. Results: Isolation and purification of methanolic extract of D. rappa stem bark yielded 14 known oligomeric resveratrol types of compounds (1-14). Results showed that isolated ampelopsin E (5) and vaticanol C (14) displayed moderate activity against human MCF-7 breast cancer and A549 lung cancer cell lines with IC50 values 14.3 and 10.7 μg/mL, respectively. Interestingly, acetate derivative of isolated laevifonol (2) and ampelopsin F (3) was found to possess potent activities towards MCF-7 cancer cell line with IC50 values 2.8 and 3.3 μg/mL, respectively, in comparison to the parental compounds that demonstrated weak activities (IC50 > 50 μg/mL). For the antibacterial assay, compounds 10 and 12 showed moderate activities towards Gram-positive bacterial strains (MIC ≤ 50 μM). Conclusion: 14 known oligomeric resveratrol types of compounds have been isolated. The activity of 14 against A549 cell line, 5 against MCF-7 cell line, and 10 and 12 against Gram-positive bacterial strains were the most promising results of this study. While the strong cytotoxicity of acetate derivative (2 and 3) against MCF-7 cell line has revealed the potential of resveratrol oligomers to be used as a template for designing new anticancer drugs.

Isobavachalcone induces cell death through multiple pathways in human breast cancer MCF-7 cells

To explore the effects of isobavachalcone (IBC) on cell death of human breast cancer MCF-7 cells and explore the possible mechanism. MCF-7 cells were treated with different concentrations of IBC, and the changes in cell proliferation were assessed using MTT assay. Apoptosis of MCF-7 cells following treatment with 10, 20, and 40 μmol/L IBC was analyzed using flow cytometry with annexin V-FITC/PI double staining and fluorescence microscopy, and the expressions of apoptosis- and autophagy-related proteins (Bax, Bcl-2, Akt, p-Akt, p62, and LC3) were detected with Western blotting. Electron microscopy was used to observe the changes in submicrostructure of the cells following treatment with 40 μmol/L IBC. JC-1 assay kit, ATP assay kit, and reactive oxygen species (ROS) kit were used to determine the effect of IBC on mitochondrial function of the cells. MTT assay showed that IBC significantly inhibited the proliferation of MCF-7 cells in a concentration- and time-dependent manner, with IC50 values of 38.46, 31.31, and 28.26 μmol/L at 24, 48, and 72 h, respectively. IBC also concentration-dependently induced apoptosis of MCF-7 cells. IBC-induced cell death was inhibited by z-VAD-fmk, a caspase inhibitor (P < 0.05), but not by the necroptosis inhibitor necrostatin-1 (Nec-1). Western blotting showed that IBC-induced MCF-7 cell apoptosis by increasing Bax expression and down-regulating the expressions of Bcl-2, Akt and p-Akt-473 (all P < 0.05). With the increase of IBC concentration, the expression of autophagy-related protein p62 and the LC3-II/I ratio increased progressively. Electron microscopy revealed the presence of autophagic bodies in IBC-treated MCF-7 cells. IBC treatment also resulted in decreased mitochondrial membrane potential and intracellular ATP level and increased ROS accumulation in MCF-7 cells (P < 0.05). IBC is capable of inducing both apoptosis and autophagy in MCF-7 cells, suggesting the potential value of IBC as a lead compound in the development of anti-breast cancer agents.

In vitro anticancer activity of Semecarpus anacadium on MCF-7 human breast cancer cells

Semecarpus anacardium (SA) is a plant native to India that is eaten as a fresh vegetable on a daily basis. The effects of SA therapy on human breast cancer MCF-7 cell lines were investigated in this study. Maceration with various solvents was used to prepare the crude extract of flavonoid fraction from SA bark, and fractionation was done using TLC and column chromatography techniques. MTT assay was used to assess the effects of the isolated flavonoid fraction of SA bark on cell viability. SA decreased MCF-7 cancer cell viability in a concentration-dependent manner, with IC50 values of 24.18+1.20 μg/mL and 12.08+1.18 μg/mL after 24 and 48 hours of treatment, respectively. Finally, SA flavonoid fraction effectively reduced MCF-7 cells, implying that SA flavonoid fraction has anticancer potential against MCF-7 cells and may be useful in the prevention and treatment of breast cancer.

Human Growth Hormone Fragment 176–191 Peptide Enhances the Toxicity of Doxorubicin-Loaded Chitosan Nanoparticles Against MCF-7 Breast Cancer Cells

IntroductionNumerous drugs with potent toxicity against cancer cells are available for treating malignancies, but therapeutic efficacies are limited due to their inefficient tumor targeting and deleterious effects on non-cancerous tissue. Therefore, two improvements are mandatory for improved chemotherapy 1) novel delivery techniques that can target cancer cells to deliver anticancer drugs and 2) methods to specifically enhance drug efficacy within tumors. The loading of inert drug carriers with anticancer agents and peptides which are able to bind (target) tumor-related proteins to enhance tumor drug accumulation and local cytotoxicity is a most promising approach.ObjectiveTo evaluate the anticancer efficacy of Chitosan nanoparticles loaded with human growth hormone hGH fragment 176–191 peptide plus the clinical chemotherapeutic doxorubicin in comparison with Chitosan loaded with doxorubicin alone.MethodsTwo sets of in silico experiments were performed using molecular docking simulations to determine the influence of hGH fragment 176–191 peptide on the anticancer efficacy of doxorubicin 1) the binding affinities of hGH fragment 176–191 peptide to the breast cancer receptors, 2) the effects of hGH fragment 176–191 peptide binding on doxorubicin binding to these same receptors. Further, the influence of hGH fragment 176–191 peptide on the anticancer efficacy of doxorubicin was validated using viability assay in Human MCF-7 breast cancer cells.ResultsIn silico analysis suggested that addition of the hGH fragment to doxorubicin-loaded Chitosan nanoparticles can enhance doxorubicin binding to multiple breast cancer protein targets, while photon correlation spectroscopy revealed that the synthesized dual-loaded Chitosan nanoparticles possess clinically favorable particle size, polydispersity index, as well as zeta potential.ConclusionThese dual-loaded Chitosan nanoparticles demonstrated greater anti-proliferative activity against a breast cancer cell line (MCF-7) than doxorubicin-loaded Chitosan. This dual-loading strategy may enhance the anticancer potency of doxorubicin and reduce the clinical side effects associated with non-target tissue exposure.

Ursolic Acid Enhances the Sensitivity of MCF-7 and MDA-MB-231 Cells to Epirubicin by Modulating the Autophagy Pathway.

Breast cancer is the leading cause of cancer death among women in the world, and its morbidity and mortality are increasing year by year. Epirubicin (EPI) is a commonly used drug for the treatment of breast cancer but unfortunately can cause cardiac toxicity in patients because of dose accumulation. Therefore, there is an urgent need for new therapies to enhance the sensitivity of breast cancer cells to EPI. In this study, we found ursolic acid (UA) can significantly improve the drug sensitivity of human breast cancer MCF-7/MDA-MB-231 cells to EPI. Next, we observed that the co-treatment of UA and EPI can up-regulate the expression of autophagy-related proteins Beclin-1, LC3-II/LC3-I, Atg5, and Atg7, and decrease the expression levels of PI3K and AKT, which indicates that the potential mechanism should be carried out by the regulating class III PI3K(VPS34)/Beclin-1 pathway and PI3K/AKT/mTOR pathway. Furthermore, we found the autophagy inhibitor 3-methyladenine (3-MA) could significantly reverse the inhibitory effect of co-treatment of UA and EPI on MCF-7 and MDA-MB-231 cells. These findings indicate that UA can dramatically enhance the sensitivity of MCF-7 and MDA-MB-231 cells to EPI by modulating the autophagy pathway. Our study may provide a new therapeutic strategy for combination therapy.

Citric acid-mediated green synthesis of selenium nanoparticles: antioxidant, antimicrobial, and anticoagulant potential applications.

Using microwave technique in the presence of citric acid, selenium nanoparticles (SeNPs) were fabricated. The morphological characteristics revealed that the spherical SeNPs with diameters ranging from 10.5 to 20 nm aggregated spherical shapes with sizes ranging from 0.67 to 0.83 mm. Moreover, the antioxidant efficacy was assessed by the DPPH radical scavenging test, which depicted that green-prepared nanoparticle at a 106.3 mg/mL dosage had the maximum scavenging capacity (301.1 ± 11.42 mg/g). Otherwise, with nanoparticle concentrations of 500 mg/ml, in vitro cell viability of SeNPs through human breast cancer MCF-7 cell lines was reduced to 61.2 ± 2.2% after 1 day of exposure. The antibacterial activity was tested against G-negative Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli), G-positive bacteria Bacillus subtilis (B. subtilis), and Staphylococcus aureus (S. aureus), which demonstrated that SeNPs had little activity against S. aureus. Still, it had the highest activity against E. coli, with a zone of inhibition (ZOI) of 25.2 ± 1.5 mm compared to 16.0 ± 0.6 mm for the standard antibiotic. Most notably, biogenic SeNPs have anticoagulant activities using activated partial thromboplastin time (aPTT) assessment. Based on previous findings, SeNPs can be used in medical aid and their cell viability, antioxidant, anticoagulant, and effects on bacteria.

Detoxification of the mycoestrogen zearalenone by Bacillus licheniformis spore CotA laccase and application of immobilized laccase in contaminated corn meal

Zearalenone (ZEN) is an estrogen-like mycotoxin posing a great threat to human and animal health. The present study showed that the induction of spore CotA laccase production of Bacillus licheniformis by ions Mn2+ and Cu2+ promoted the ZEN-degrading ability of the strain. The spore CotA laccase heterologously expressed in Escherichia coli was capable of directly oxidizing ZEN with Km, Kcat and Vmax values of 90.43 μg mL−1, 0.11s−1 and 35.48 μg min−1 mg−1, respectively. The use of 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), acetosyringone, ferulic acid, caffeic acid and methyl syringate as redox mediators could enhance ZEN degradation by CotA laccase. The human breast cancer MCF-7 cell proliferation assay showed that CotA laccase and CotA laccase-mediator systems eliminated the estrogenicity of ZEN. Immobilized CotA laccase onto chitosan microspheres exhibited higher thermal stability over free CotA laccase, maintaining about 87% of its initial activity after heat treatment at 80 °C for 30 min. The removal rate of ZEN in corn meal by free and immobilized CotA laccase was 70% and 90%, respectively. Meanwhile, immobilized enzyme could achieve a 54% ZEN degradation in corn meal after three consecutive cycles. Thus, CotA laccase could be a promising biocatalyst for detoxification of ZEN in corn meal.

Fabrication of Hyaluronic Acid Surface Modified Solid Lipid Nanoparticles Loaded with Imatinib Mesylate for Targeting Human Breast Cancer MCF-7 Cells

Fabrication of Hyaluronic Acid Surface Modified Solid Lipid Nanoparticles Loaded with Imatinib Mesylate for Targeting Human Breast Cancer MCF-7 Cells

Retusone A, a Guaiane-Type Sesquiterpene Dimer from Wikstroemia retusa and Its Inhibitory Effects on Histone Acetyltransferase HBO1 Expression.

Retusone A (1), a new sesquiterpene dimer consisting of two guaiane-type sesquiterpenoids, and oleodaphnal (2) were isolated from heartwood of Wikstroemia retusa (Thymelaeaceae). The planar structure of 1 was elucidated on the basis of HRESIMS and NMR spectroscopic data, and the relative stereochemistry was established by X-ray diffraction analysis. The absolute configuration of 1 was determined by electronic circular dichroism. Compound 1 suppressed luciferase reporter gene expression driven by the HBO1 (histone acetyltransferase binding to ORC1) gene promoter in human breast cancer MCF7 cells. Compound 1 also decreased the expression of endogenous HBO1 mRNA and protein, and inhibited proliferation of the cells. These results suggest that retusone A (1), which has a unique dimeric sesquiterpenoid structure with inhibitory activity against HBO1 expression, may contribute to the development of a novel therapeutic candidate for the treatment of breast cancer.

Cynometra cauliflora essential oils loaded-chitosan nanoparticles: Evaluations of their antioxidant, antimicrobial and cytotoxic activities

Nanoencapsulation has appeared as an alternative approach to protect the bioactive constituents of essential oils (EOs) and to improve their properties. In this study, Cynometra cauliflora essential oils (CCEOs) were nanoencapsulated in chitosan nanoparticles (CSNPs) using an emulsion-ionic gelation technique. Transmission electron microscopy (TEM) images illustrated a well dispersion and spherical shape of C. cauliflora EOs-loaded chitosan nanoparticles (CCEOs-CSNPs) with an average size of less than 100 nm. In addition to that, Fourier transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS) and X-ray diffraction (XRD) analyses revealed the success of CCEOs nanoencapsulation. The encapsulation efficiency (EE) was in the range of 38.83% to 44.16% while the loading capacity (LC) reached 32.55% to 33.73%. The antioxidant activity (IC50) of CCEOs-CSNPs was ranged from 21.65 to 259.13 μg/mL when assessed using DPPH radical scavenging assay. CCEOs-CSNPs showed an appreciable antimicrobial effects on diabetic wound microorganisms. Notably, cytotoxic effects against human breast cancer MCF-7 and MDA-MB-231 cells recorded IC50 of 3.72–17.81 μg/mL and 16.24–17.65 μg/mL, respectively, after 72 h treatment. Interestingly, no cytotoxicity against human breast normal MCF-10A cells was observed. Thus, nanoencapsulation using CSNPs could improve the properties of CCEOs in biomedical related applications.

Effects of tumor-derived extracellular vesicles on T cell fate and function

Abstract Immunosuppressive microenvironments created by malignant tumors represent a major obstacle for effective anti-tumor immunity. There are multiple immunosuppressive mechanisms operative in the tumor microenvironment (TME) that interfere with T cell function. Extracellular vesicle (EV) in tumor microenvironment is an important way employed by tumor cells to control T cell fate and function. However, the mechanisms responsible for EV-mediated T cell suppression and dysfunction are still largely unknown. Here, we purified EVs from human melanoma A375 and breast cancer MCF7 cell lines and then explored EV impacts on T cell function and fate in anti-tumor immunity. We found that tumor derived EVs can suppress T cell proliferation and induce T cell senescence, which is a novel and fundamentally important phenomenon that contributes to tumor-induced immunosuppression in the TME. Mechanistically, tumor derived EVs promoted DNA damage responses and cell cycle arrest of T cell that induces T cell senescence. Blockage of DNA damage signaling in T cells via the specific inhibitor KU55933 can prevent T cell senescence and recover T cell impaired effector functions. Importantly, we further performed in vivo studies to investigate that tumor-derived EVs can induce T cell suppression and senescence, which can also be reversed by specific signaling inhibitors or/and targeting signaling molecules via the CRISPR knockdown strategy. These studies facilitate a better understanding of the novel suppressive mechanism utilized by the TME and provide insights relevant to the development of effective strategies to reverse immune suppression for enhanced anti-tumor immunity. Supported by the grants from the Melanoma Research Alliance and National Institutes of Health.

NNav1.5 expression is associated with glutamate level in breast cancer cells

BackgroundGlutamate and voltage-gated sodium channels, both have been the target of intense investigation for its involvement in carcinogenesis and progression of malignant disease. Breast cancer with increased level of glutamate often metastasize to other organs (especially bone), whilst re-expression of ‘neonatal’ Nav1.5, nNav1.5 in breast cancer is known to promote cell invasion in vitro, metastasis in vivo and positive lymph node metastasis in patients.MethodsIn this study, the role of nNav1.5 in regulating glutamate level in human breast cancer cells was examined using pharmacological approach (VGSCs specific blocker, TTX, glutamate release inhibitor, riluzole and siRNA-nNav1.5). Effect of these agents were evaluated based on endogenous and exogenous glutamate concentration using glutamate fluorometric assay, mRNA expression of nNav1.5 using qPCR and finally, invasion using 3D culture assay.ResultsEndogenous and exogenous glutamate levels were significantly higher in aggressive human breast cancer cells, MDA-MB-231 cells compared to less aggressive human breast cancer cells, MCF-7 and non-cancerous human breast epithelial cells, MCF-10A. Treatment with TTX to MDA-MB-231 cells resulted in significant reduction of endogenous and exogenous glutamate levels corresponded with significant suppression of cell invasion. Subsequently, downregulation of nNav1.5 gene was observed in TTX-treated cells.ConclusionsAn interesting link between nNav1.5 expression and glutamate level in aggressive breast cancer cells was detected and requires further investigation.

Atractylodin induces oxidative stress-mediated apoptosis and autophagy in human breast cancer MCF-7 cells through inhibition of the P13K/Akt/mTOR pathway.

This study aimed to determine the apoptosis and autophagy-inducing mechanism of atractylodin in human breast cancer MCF-7 cells. The molecular mechanism of anticancer activity of atractylodin was confirmed by assessing the levels of reactive oxygen species (ROS) level, lipid peroxidation (LPO), antioxidants activity, dual staining, and comet assay. Moreover, cleaved caspases 3, 8, and 9, and signaling proteins, such as p53, Bcl-2, and Bax, phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin(P13K/Akt/mTOR), LC3I andLC3II, and beclin-1 were analyzed. In MCF-7 cells treated with atractylodin, the concentration-dependent toxicity, increased LPO, increased production of ROS, and decreased activity of superoxide dismutase, catalase, and glutathione peroxidasewere observed. In MCF-7 cells, atractylodin administration decreased Bcl-2 expression while activating the expression of p53, Bax, cleaved caspase-3, caspase-8, and caspase-9 apoptotic members. Furthermore, atractylodin blocked the P13K/Akt/mTOR signaling pathway, increased the conversion of LC3I to its lipidated form of LC3II, and increased beclin-1 expression, whereas downregulated the p62 expression in MCF-7 cells. As a result, altering apoptotic and autophagy-related biomarkers, atractylodin triggered apoptosis and autophagy in MCF-7 cells. As a result, atractylodin could be utilized to treat human breast cancer after the proper clinical trial.