MDA-MB231 Cells Research Articles

Effect of prevalent polychlorinated biphenyls (PCBs) food contaminant on the MCF7, LNCap and MDA-MB-231 cell lines viability and PON1 gene expression level: proposed model of binding.

Polychlorinated biphenyls (PCBs) are a group of synthetic organic chlorine compounds known as an organic pollutant in food sources, which play important roles in malignancies. The present study aimed to investigate the direct effects of prevalent PCBs in food in hormone-responsive and non-responsive cell lines. In the current study, MCF-7, LNCap, and MDA-MB231 cell lines were treated with serial concentrations (0.001-100μM) of PCBs for 48h and cell viability assessment was performed using MTT assay. The best concentration then applied and the expression level of PON1 was evaluated using real-time PCR. Besides, molecular docking was performed to determine the binding mechanism and predicted binding energies of PBCs compounds to the AhR receptor. Unlike MCF-7 and LNCap cells, the viability of MDA-MB231 cells did not significantly change by different concentrations of PCBs. Meanwhile, quantitative gene expression analysis showed that the PON1 was significantly more expressed in MCF-7 and LNCap lines treated with PCB28 and PCB101. However, the expression level of this gene in other groups and also MDA-MB231cells did not demonstrate any significantly change. Also, the results of molecular docking showed that PBCs had steric interaction with AhR receptor. Current results showed that despite of hormone non-responsive cells the PCBs have a significant positive effect on hormone-responsive cell. Therefore, and regarding to the existence of PCBs contamination in food there should be serious concern about their impact on the prevalence of different malignancies which certainly should result in a standard limit for this material. This study aimed to investigate the direct effects of prevalent PCBs in food in hormone-responsive and non-responsive cell lines. Cell lines were treated with serial concentrations of PCBs and cell viability assessment was performed using MTT assay. The expression level of PON1 was evaluated using real-time PCR. Molecular docking was performed to determine the binding mechanism and predicted binding energies of PBCs compounds to the AhR receptor. PCBs contamination in food there should be serious concern about their impact on the prevalence of different malignancies which certainly should result in a standard limit for this material.

A Novel Thienopyrimidine Analog, TPH104, Mediates Immunogenic Cell Death in Triple-Negative Breast Cancer Cells.

Simple SummaryTriple-negative breast cancer (TNBC) is the most lethal and aggressive subtype of breast cancer that lacks an estrogen receptor, the progesterone receptor and the human epidermal growth factor receptor 2 (HER2), making it unsuitable for hormonal- or HER2-based therapy. TNBC is known for its higher relapse rate, poorer prognosis and higher rate of metastasis compared to non-TNBC because although patients initially respond to chemotherapy that kills cancer cells through a form of programmed cell death called apoptosis, they later develop chemoresistance and stop responding to the treatment, accounting for one fourth of all breast cancer deaths. In this study, we report a novel compound, TPH104, that elicits a unique, non-apoptotic cell death in TNBC cells. Upon treatment with TPH104, TNBC cells swell and burst, releasing immunogenic markers that alert and activate the immune system to further recognize and attack the neighboring breast cancer cells.Enhancing the tumor immunogenic microenvironment has been suggested to circumvent triple-negative breast cancer (TNBC) resistance and increase the efficacy of conventional chemotherapy. Here, we report a novel chemotherapeutic compound, TPH104, which induces immunogenic cell death in the TNBC cell line MDA-MB-231, by increasing the stimulatory capacity of dendritic cells (DCs), with an IC50 value of 140 nM. TPH104 (5 µM) significantly increased ATP levels in the supernatant and mobilized intracellular calreticulin to the plasma membrane in MDA-MB-231 cells, compared to cells incubated with the vehicle. Incubating MDA-MB-231 cells for 12 h with TPH104 (1–5 µM) significantly increased TNF-α mRNA levels. The supernatants of dying MDAMB-231 cells incubated with TPH104 increased mouse bone marrow-derived DC maturation, the expression of MHC-II and CD86 and the mRNA expression of TNF-α, IL-6 and IL-12. Overall, these results indicate that TPH104 induces immunogenic cell death in TNBC cells, in part, by activating DCs.

Resveratrol Enhances Inhibition Effects of Cisplatin on Cell Migration and Invasion and Tumor Growth in Breast Cancer MDA-MB-231 Cell Models In Vivo and In Vitro.

Triple-negative breast cancer (TNBC) is a refractory type of breast cancer that does not yet have clinically effective drugs. The aim of this study is to investigate the synergistic effects and mechanisms of resveratrol combined with cisplatin on human breast cancer MDA-MB-231 (MDA231) cell viability, migration, and invasion in vivo and in vitro. In vitro, MTS assays showed that resveratrol combined with cisplatin inhibits cell viability as a concentration-dependent manner, and produced synergistic effects (CI < 1). Transwell assay showed that the combined treatment inhibits TGF-β1-induced cell migration and invasion. Immunofluorescence assays confirmed that resveratrol upregulated E-cadherin expression and downregulated vimentin expression. Western blot assay demonstrated that resveratrol combined with cisplatin significantly reduced the expression of fibronectin, vimentin, P-AKT, P-PI3K, P-JNK, P-ERK, Sma2, and Smad3 induced by TGF-β1 (p < 0.05), and increased the expression of E-cadherin (p < 0.05), respectively. In vivo, resveratrol enhanced tumor growth inhibition and reduced body weight loss and kidney function impairment by cisplatin in MDA231 xenografts, and significantly reduced the expressions of P-AKT, P-PI3K, Smad2, Smad3, P-JNK, P-ERK, and NF-κB in tumor tissues (p < 0.05). These results indicated that resveratrol combined with cisplatin inhibits the viability of breast cancer MDA231 cells synergistically, and inhibits MDA231 cells invasion and migration through Epithelial-mesenchymal transition (EMT) approach, and resveratrol enhanced anti-tumor effect and reduced side of cisplatin in MDA231 xenografts. The mechanism may be involved in the regulations of PI3K/AKT, JNK, ERK and NF-κB expressions.

Migration and shape of cells on different interfaces* *Project supported by the National Natural Science Foundation of China (Grant Nos. 11774394 and 11704404), the Chinese Academy of Sciences (CAS), and the Key Research Program of Frontier Sciences of CAS (Grant No. QYZDB-SSW-SYS003).

Impacts of microenvironments on cell migration have been reported in various interaction modes. A rapid tumor metastasis occurs along topological interfaces in vivo, such as the interface between the blood vessels and nerves. In this work, we culture MDA-MB231 cells at dish-liquid, dish-hydrogel, and hydrogel-liquid interfaces, respectively, to study how these different interfaces influence cell dynamics and morphology. Our results show that the migration mode of cells changes from an amoeboid motion to a mesenchymal motion but their speed do not change obviously if the interface changes from hydrogel-liquid to dish-liquid. In contrast, the migration mode of cells at a dish-hydrogel interface maintains as a mesenchymal motion, whereas their speed increases significantly.

Identification of the effects of acid-resistant Lactobacillus casei metallopeptidase gene under colon-specific promoter on the colorectal and breast cancer cell lines.

Objective(s):Anti-tumor effects of Lactobacilli as normal flora have been described. In a previous study, we identified a protein isolated from the bacterium Lactobacillus casei ATCC 39392 in acidic pH conditions named metallopeptidase. Therefore, we decided to evaluate the effect of the recombinant plasmid coding metallopeptidase protein on the inhibition, proliferation, or apoptosis of the colorectal and breast cancer cell lines. Materials and Methods:Identified metallopeptidase gene of L. casei under the specific colon cancer promoter was transferred to the Human SW480 and MDA-MB231 cells. Cell viability was evaluated in these two cancer cell lines via MTT assay, apoptotic changes, and expression level of p53 and MAP2K1 genes in comparison with healthy blood cells as a control group. Results:Viability of SW480 and MDA-MB231 cells was identified at 25% and 7%, respectively. An increase in apoptotic cell death in the SW480 cell line was observed as revealed by Tunnel staining. The expression assay of TP53 and MAP2K1 genes showed that MPL protein altered gene expression in a cell type-specific manner. Tunnel analyses showed that the pronounced cytotoxic effect of pEGFP-C2/MPL plasmid on SW480 cells was mediated through apoptosis. Conclusion:These results suggest that endogenous recombinant MPL under colon specific promoter inhibits the proliferation of SW480 colorectal cancer cells by increase in MAP2K1 and P53 activation. L. casei metallopeptidase under the same circumstances could not affect the growth rate and viability of MDA-MB231 breast cancer cells in vitro.

Design, cytotoxic effects on breast cancer cell line (MDA-MB 231), and molecular docking of some maleimide-benzenesulfonamide derivatives

A group of novel maleimide-benzenesulfonamide derivatives 3a-d was designed and synthesized for their evaluation as a potential anti-breast cancer agent. The structures of these derivatives were confirmed by their 1H, 13C NMR, Mass, FT-IR spectral data, and melting points. The cytotoxic activity (in vitro) of the selected molecules against MDA-MB231 ​cell line was evaluated by MTT method. Among them, compounds 3a and 3d exhibited a significant cytotoxicity with the IC50 value of 1.61 and 1.26 ​μM, respectively, whereas compounds 3b and 3c showed a moderate cytotoxicity with IC50 values of 0.45 and 1.12 ​μM, respectively against MDA-MB231 ​cells. Docking modeling of the synthesized compounds 3a-d into binding sites of human aromatase protein (PDB ID: 4GL7) was performed to investigate if these derivatives possess analogous binding mode to breast cancer proteins. Docking results showed these compounds have efficient interactions such as hydrogen bonding, Van der Waals interactions, and hydrophobic interactions with the active site residues of the aromatase protein (PDB ID: 4GL7). The low binding energies and a number of hydrogen bonding indicated that the maleimide-benzenesulfonamide derivatives might be considered as a promising anti-breast cancer agent with further developments in drug discovery.

Dihydrostilbene glycosides from Camellia sinensis var. assamica and their cytotoxic activity

Three undescribed dihydrostilbene glycosides, 3,5-dihydroxyldihydrostilbene 4′-O-[6′′-O-(4′′′-hydroxylbenzoyl)]-β-D-glucopyranoside (1), 3,5-dihydroxyldihydrostilbene 4′-O-(6′′-O-galloyl)-β-D-glucopyranoside (2), and 3,5-dihydroxyldihydrostilbene 4′-O-[6′′-O-(3′′′,4′′′-dimethoxyl)galloyl]-β-D-glucopyranoside (3), and seven known compounds, kaempferol 3-O-β-D-glucopyranoside (4), isoquercitrin (5), kaempferol 3-O-α-L-rhamnoside (6), quercitrin (7), (6S,9R)-roseoside (8), (-)-epicatechin 3-O-gallate (9), and (-)-epigallocatechin 3-O-gallate (10) have been isolated from the methanol extract of the leaves of Camellia sinensis var. assamica (J.W.Mast.) Kitam. (synnonym of Camellia assamica (Mast.) H.T.Chang) (Theaceae). Their structures were elucidated by spectroscopic methods (1 D-, 2 D-NMR) and mass spectra. All compounds were evaluated for cytotoxic activity against human oral cancer (CAL27) and human breast cancer (MDAMB231) cell lines. Compound 10 showed significant cytotoxic activity against CAL27 and MDAMB231 cell lines with IC50 values of 9.78 ± 0.25 and 3.27 ± 0.18 μM, respectively, compared to those of positive control, capecitabine (IC50 values of 8.20 ± 0.75 and 5.20 ± 0.89 μM).

Phytochemical, elemental, physico-chemical, HPTLC and anticancer investigations of Ceropegia spiralis Wight. Tuber extracts

Phytochemical constituents are responsible for medicinal activity of plant species. Hence in the present study Qualitative and quantitative phytochemical screening, Physicochemical, elemental analysis and anticancer activity of Ceropegia spiralis tuber extracts were carried out. Qualitative and quantitative phytochemical analysis of tuber aqueous extract confirm the presence of various secondary metabolites like saponins, triterpenoids, steroids, tannins, alkaloids, flavonoids and phenols. The results suggest that the phytochemical properties for curing various ailments and possess potential anticancer, antimicrobial and antioxidant activities leads to the isolation of new and novel compounds. Physicochemical studies reveals that the dry matter 96.23%; followed by Water soluble extractive 14.28 %. Elemental analysis reveals Nitrogen 3.4 % followed by Zinc 218.9%. Along with this, macro and micro elements which are essential for maintaining the animal body were also determined quantitatively. Anticancer activity of C. spiralis exhibited potential towards MDAMB-231 (human breast cancer) cell lines, shows 78.30% cell death with cell viability 21.70% at 100 µg/ml. The presence of various bioactive compounds confirms the application of C. spiralis against many ailments by the traditional practitioners.

Saurobacciosides A - C: three new glycosides from Sauropus bacciformis with their cytotoxic activity

Three new glycosides, named as saurobaccioside A (1), saurobaccioside B (2), saurobaccioside C (3), together with five known magastigmanes, canangaionoside (4), (6S,9S)-roseoside (5), cucumegastigmane I (6), icariside B5 (7), linarionoside A (8) were isolated from the whole plant of Sauropus bacciformis (L.) Airy Shaw. Their structures were established by extensive spectroscopic analysis (UV, IR, HR-ESI-MS and NMR) and by comparison of the spectral data with those reported in the literature. The absolute configurations of compounds 2 and 3 were elucidated by experimental CD spectra. Compounds 1-8 were screened their cytotoxic activities towards CAL27 and MDAMB231 cancer cell lines. Compound 1 exhibited significant cytotoxic activity towards CAL27 and MDAMB231 cell lines with IC50 values of 3.21 ± 0.23 and 4.75 ± 0.17 µM, respectively, which were smaller than those of positive control capecitabine (IC50: 8.20 ± 0.75 and 5.20 ± 0.89 µM). Other compounds (2-8) were inactive.

Influence of the obstacles on dielectrophoresis-assisted separation in microfluidic devices for cancerous cells

The development of a separation lab-on-a-chip device based on the particle size is critical in the early detection of circulating tumor cells. In this work, combining alternating current dielectrophoresis and drag force are used to precisely separate MDA-MB231 and red blood cells in the microfluidic platform. Four microfluidic devices with sidewall electrodes and different obstacle shapes were employed to both generate non-uniform fields inside the channels and condense the particles near the electrodes. Besides that, dividing the particles inlet into two main channels enable fast and accurate separation inside the device, and then particles deflected to the different outlets. Choosing the frequency is critical in the magnitude and sign of the dielectrophoresis force. Negative dielectrophoresis force was acting on both cancerous and red blood cells when the frequency was 10 kHz. Finite element simulation was conducted to predict the particle trajectories and revealed that the device with rectangular obstacles had the higher electric field non-uniformity when the voltage applied was set to 6 V and − 6 with the frequency of 10 kHz, Simulation represented 100% separation efficiency and purity for cancerous cells in the specific outlets. Decreasing the voltage from 6 V and − 6 V to 5 V and − 5 V could reduce the separation efficiency to 46%. The appropriate value of voltage (6 V, − 6 V) was applied to the electrodes resulting in the lower Joule heating and sufficient dielectrophoresis forces.

Curcumin Encapsulated in Crosslinked Cyclodextrin Nanoparticles Enables Immediate Inhibition of Cell Growth and Efficient Killing of Cancer Cells.

The efficiency of anti-cancer drugs is commonly determined by endpoint assays after extended incubation times, often after days. Here we demonstrate that curcumin encapsulated in crosslinked cyclodextrin nanoparticles (CD-NP) acts extremely rapidly on cell metabolism resulting in an immediate and complete inhibition of cell growth and in efficient cancer-cell killing only few hours after incubation. This early onset of anti-cancer action was discovered by live-cell high-throughput fluorescence microscopy using an environmental stage. To date, only very few examples of covalently crosslinked nanoscale CD-based (CD-NP) drug carriers exist. Crosslinking cyclodextrins enables the adsorption of unusually high payloads of hydrophobic curcumin (762 µg CC/mg CD-NP) reflecting a molar ratio of 2.3:1 curcumin to cyclodextrin. We have investigated the effect of CD-NP encapsulated curcumin (CD-CC-NP) in comparison to free, DMSO-derived curcumin nanoparticles (CC-NP) on 4 different cell lines. Very short incubations times as low as 1 h were applied and cell responses after medium change were subsequently followed over two days. We show that cell proliferation is inhibited nearly immediately in all cell lines and that a cell- and concentration dependent cancer-cell killing occurs. Anti-cancer effects were similar with free and encapsulated curcumin, however, encapsulation in CD-NP drastically extends the long-term photostability and anti-cancer activity of curcumin. Curcumin-sensitivity is highest in HeLa cells reaching up to 90% cell death under these conditions. Sensitivity decreased from HeLa to T24 to MDA MB-231 cells. Strikingly, the immortalized non-cancerous cell line MCF-10A was robust against curcumin concentrations that were highly toxic to the other cell lines. Our results underline the potential of curcumin as gentle and yet effective natural anti-cancer agent when delivered solvent-free in stabilizing and biocompatible drug carriers such as CD-NP that enable efficient cellular delivery.

Design, synthesis of 4-[2-(substituted phenyl) hydrazono]-3-(1-hydroxyethyl)-1-phenyl/methyl-3,4-dihydroquinolin-2(1H)-one derivatives and evaluation of their in vitro tyrosine kinase inhibitor activity

The present investigation deals with molecular docking, synthesis, characterization, and evaluation of in vitro tyrosine kinase inhibitor activity of a series of 4-[2-(substituted phenyl) hydrazono]-3-(1-hydroxyethyl)-1-phenyl/methyl- 3,4-dihydroquinolin-2(1 H )-one derivatives {III-a(1-12)/III-b(1-12)}. Molecular docking studies of the title compounds were carried out using Molegro Virtual Docker (MVD-2013, 6.0) software. The MolDock scores of the derivatives ranged from (−66.508) to (−101.274); whereas the MolDock score of standard 4-anilinoquinazoline ligand was found to be (−105.219). Most of the synthesized qunolin-2-one derivatives showed better affinity towards EGFRK protein as compared to standard drug imatinib (−104.253). All the synthesized compounds were satisfactorily characterized by physical and spectral analysis (UV, IR, 1 H NMR and 13 C NMR and mass spectral data). Twelve derivatives were tested for their in vitro tyrosine kinase inhibitor activity using MDA-MB cell line. Compound 4-[2-(4-bromophenyl)hydrazono]-3-(1-hydroxyethyl)-1- methyl- 3,4-dihydroquinolin-2(1 H )-one (III-b4) was found to be the most cytotoxic compound as compared to other synthesized derivatives, with IC 50 value of 0.0515 μM against MDA- MB cell line.

Folic Acid-Functionalized Nanomedicine: Folic Acid Conjugated Copolymer and Folate Receptor Interactions Disrupt Receptor Functionality Resulting in Dual Therapeutic Anti-Cancer Potential in Breast and Prostate Cancer

We have previously reported on a functionalized folic acid (FA) conjugated poly(styrene-alt-maleic anhydride) (SMA) via biological linker 2,4-diaminobutyric acid (DABA) (FA-DABA-SMA) copolymer. This biocompatible nanocopolymer self-assembles in a pH-dependent manner, providing stimuli responsiveness, active targeting, and extended release of hydrophobic chemotherapeutic agents and effectively penetrates the inner core of 3-dimensional cancer spheroid models. The empty FA-DABA-SMA decreased tumor spheroid volume, revealing a previously unknown mechanism of action. Here, we investigated the potential mechanism of the small (20 kDa) and large (350 kDa) FA-DABA-SMA empty copolymers affecting the folic acid receptor alpha (FRα) signaling properties in breast and prostate cancer cell lines. Microscopic imaging, immunocytochemistry, flow cytometry, Caspase 3/7 apoptosis assays, Incucyte live cell tracking, the scratch wound assay, the water-soluble tetrazolium salt-1 (WST-1) cell viability assay, morphologic changes, and Western blot for the expression levels of FRα on the cell surface were used on MDA MB-231 and MCF-7 breast and DU-145 prostate cancer cell lines. The findings indicate that FA-DABA-SMA increases FRα expression levels in breast MDA MB-231 cancer cells and then disrupts FR signaling by reducing HES1 and NOTCH1 protein expression levels. Also, FA-DABA-SMA induces apoptosis and further causes a change in the MDA MB-231 cells' morphology and significantly reduces their ability to migrate in a scratch wound assay. Collectively, these findings provide a novel insight into the functionalized FA-DABA-SMA copolymer. The 350 and 20 kDa copolymers actively target FRα to initialize internationalization. However, only the large size and sheet-shaped 350 kDa copolymers disrupt FRα signaling. The significance of these novel findings reveals that the copolymer's intracellular activity is critically dependent on the size and structural shape. This report offers novel therapeutic insight into a dual mechanism of the FA-DABA-SMA copolymer for its therapeutic potential to treat cancer.

Cytotoxic and antiproliferative potential of methanolic extracts of Asparagus racemosus in MDAMB231 cells

The present study was carried out to evaluate the cytotoxic and antiproliferative activity of methanolic extract of Asparagus racemosus in MDAB-231 cells. The qualitative phytochemical analysis of the extract revealed the presence of steroids, alkaloids, flavonoids, glycosides, saponins and diterpenes. MDAMB-231 cells were maintained in RPMI media containing 10 per cent serum and 1 per cent antibiotic and antimycotic solution. The cells were harvested and seeded to 96 well plate at 5×103 cells/mL, incubated for 24 hours at 37˚C with 5 per cent CO2. The cells were treated with 160, 80, 40, 20 and 10µg/mL of the extract for 24 hours and viability was accessed using MTT Assay. Also cells were plated in 6 well plates at concentrations 3×105 cells/mL and exposed to 80, 40, 20 and 10µg/mL of the extract of Asparagus racemosus for acridine orange ethidium bromide (AO/EB) staining. There was a dose dependent decrease in viability of cells exposed to methanolic extracts of Asparagus racemosus with a viability of 15.55+0.193 per cent for cells treated with 160µg/mL. The IC50 was found to be 91.36+ 0.87 µg/mL. The AO/EB staining revealed that most of the cells were dead in extract treated with 80µg/mL where more than 90 per cent of cells were live when treated with 10µg/mL of the extract.

Comparative stemness and differentiation of luminal and basal breast cancer stem cell type under glutamine-deprivation.

Glutamine (gln) metabolism has emerged as a cancer therapeutic target in past few years, however, the effect of gln-deprivation of bCSCs remains elusive in breast cancer. In this study, effect of glutamine on stemness and differentiation potential of bCSCs isolated from MCF-7 and MDAMB-231 were studied. We have shown that bCSCs differentiate into CD24+ epithelial population under gln-deprivation and demonstrated increased expression of epithelial markers such as e-cadherin, claudin-1 and decreased expression of mesenchymal protein n-cadherin. MCF-7-bCSCs showed a decrease in EpCAMhigh population whereas MDAMB-231-bCSCs increased CD44high population in response to gln-deprivation. The expression of intracellular stem cell markers such sox-2, oct-4 and nanog showed a drastic decrease in gene expression under gln-deprived MDAMB-231-bCSCs. Finally, localization of β-catenin in MCF-7 and MDAMB-231 cells showed its accumulation in cytosol or perinuclear space reducing its efficiency to transcribe downstream genes. Conclusively, our study demonstrated that gln-deprivation induces differentiation of bCSCs into epithelial subtypes and also reduces stemness of bCSCs mediated by reduced nuclear localization of β-catenin. It also suggests that basal and luminal bCSCs respond differentially towards changes in extracellular and intracellular gln. This study could significantly affect the gln targeting regimen of breast cancer therapeutics.

Genome Mining Reveals a Multiproduct Sesterterpenoid Biosynthetic Gene Cluster in Aspergillus ustus.

Genome mining of Aspergillus ustus 094102 enabled the discovery of a multiproduct bifunctional terpene synthase (BTS), AuAS. Heterologous expression of AuAS led to the discovery of five new sesterterpenes, and coexpression of the upstream CYP450 monooxygenase (AuAP450) generated four new sesterterpene alcohols. Additionally, aspergilol A showed cytotoxic activities against MCF-7, MDA-MB231, and HepG2 cancer cells (IC50 21.20-48.76 μM), and aspergilol B exhibited a cytotoxic effect on MCF-7 cells (IC50 27.41 μM).

Preparation and characterization of nanocurcumin based hybrid virosomes as a drug delivery vehicle with enhanced anticancerous activity and reduced toxicity

The present study represents a formulation of nanocurcumin based hybrid virosomes (NC-virosome) to deliver drugs at targeted sites. Curcumin is a bioactive component derived from Curcuma longa and well-known for its medicinal property, but it exhibits poor solubility and rapid metabolism, which led to low bioavailability and hence limits its applications. Nanocurcumin was prepared to increase the aqueous solubility and to overcome all the limitations associated with curcumin. Influenza virosomes were prepared by solubilization of the viral membrane with 1,2-distearoyl-sn-glycerol-3-phosphocholine (DSPC). During membrane reconstitution, the hydrophilic nanocurcumin was added to the solvent system, followed by overnight dialysis to obtain NC-virosomes. The same was characterized using a transmission electron microscope (TEM) and scanning electron microscope (SEM), MTT assay was used to evaluate it's in vitro-cytotoxicity using MDA-MB231 and Mesenchyme stem cells (MSCs). The results showed NC-virosomes has spherical morphology with size ranging between 60 and 90 nm. It showed 82.6% drug encapsulation efficiency. The viability of MDA-MB231 cells was significantly inhibited by NC-virosome in a concentration-dependent manner at a specific time. The IC50 for nanocurcumin and NC-virosome was 79.49 and 54.23 µg/ml, respectively. The site-specific drug-targeting, high efficacy and non- toxicity of NC-virosomes proves its future potential as drug delivery vehicles.

Multi-Targeted Anticancer Activity of Imidazolate Phosphane Gold(I) Compounds by Inhibition of DHFR and TrxR in Breast Cancer Cells.

A class of phosphane gold(I) compounds, made of azoles and phosphane ligands, was evaluated for a screening on the regards of Breast Cancer cell panels (BC). The compounds possess N-Au-P or Cl-Au-P bonds around the central metal, and they differ for the presence of aprotic or protic polar groups in the azoles and/or the phosphane moieties to tune their hydrophilicity. Among the six candidates, only the compounds having the P-Au-N environment and not displaying neither the hydroxyl nor carboxyl groups in the ligands were found active. The compounds were screened by MTT tests in SKBR3, A17, and MDA-MB231 cancer cells, and two compounds (namely the 4,5-dicyano-imidazolate-1yl-gold(I)-(triphenylphosphane, 5, and 4,5-dichloro-imidazolate-1yl-gold(I)-triphenylphosphane, 6) were found very cytotoxic, with the most active with an IC50 value of 3.46 μM in MDA-MB231 cells. By performing enzymatic assays in the treated cells lysates, the residual enzymatic activity of dihydrofolate reductase (DHFR) has been measured after cell treatment for 4 or 12 h in comparison with control cells. Upon 12 h of treatment, the activity of DHFR was significantly reduced in both SKBR3 and A17 cells by compounds 5 and 6, but not in human MDA-MB231 cells; interestingly, it was found remarkably high after 4 h of treatment, revealing a time dependence for the DHFR enzymatic assays. The DHFR inhibition data have been compared to those for the thioredoxin reductase (TrxR), the most recognized molecular target for gold compounds. For this latter, similar residual activities (i.e., 37 and 49% for the match of SKBR3 cells and compound 5 or 6, respectively) were found. Binding studies on the regards of ct-DNA (calf-thymus-DNA) and of plasma transporters proteins, such as BSA (bovine serum albumin) and ATF (apo transferrin), were performed. As expected for gold compounds, the data support strong binding to proteins (Ksv values range: 1.51 ÷ 2.46 × 104 M−1) and a weaker interaction with ct-DNA's minor groove (Ksv values range: 1.55 ÷ 6.12 × 103 M−1).

On improving the physiological stability of curcuminoids: Curcumininoid-silver nanoparticle complex as a better and efficient therapeutic agent

Curcumin, together with its analogs, namely, demethoxycurcumin and bisdemethoxycurcumin, are phenolic pigments extracted from turmeric. These curcuminoids are considered to be a potent antioxidants possessing a multitude of properties including anti-inflammatory, antitumor, anti-HIV and antimicrobial properties. One of the serious drawbacks of these curcuminoids as therapeutic agent is their fast degradation at physiological pH leading to poor bioavailability in the biological milieu. With a view to address this stability issue we synthesized curcuminoid-silver nanoparticle (CUR-AgNP) complexes, utilizing curcuminoids as reducing agents. The stability of individual curcuminoids in the CUR control and in the CUR-AgNP complex was compared after incubation in phosphate buffer of pH 7.4. Interestingly the initial degradation of curcuminoids at 4 h was only 9.6% in the CUR-AgNP complex when compared to 49.6% degradation observed in the CUR control. Among the three curcumin analogs, bisdemethoxycurcumin showed better pH stability compared to curcumin and demethoxycurcumin whereas all the three curcuminoids exhibited the same degradation pattern in the case of the CUR-AgNP complex. In vitro studies with the CUR-AgNP complex showed excellent antibacterial property against P. aeruginosa. CUR-AgNP complex also exhibited good cytotoxic effect on MDA-MB231 cells. The storage stability of CUR-AgNP complex, kept as a suspension at room temperature (∼28 oC), was tested by measuring its hydrodynamic size and zeta potential. It was found that after 340 days of storage the particle size (Z-average) and zeta potential were 65.5 ± 1.1 nm and -22.9 ± 0.2 mV, respectively, indicating that the nanoparticle suspension was stable The results obtained in the present study are promising and could be extended to improve the therapeutic efficacy of curcuminoids in the biological pH conditions.

Mangifera indica Extracts as Novel PKM2 Inhibitors for Treatment of Triple Negative Breast Cancer.

Pyruvate kinase (PK), a key enzyme that determines glycolytic activity, has been known to support the metabolic phenotype of tumor cells, and specific pyruvate kinase isoform M2 (PKM2) has been reported to fulfill divergent biosynthetic and energetic requirements of cancerous cells. PKM2 is overexpressed in several cancer types and is an emerging drug target for cancer during recent years. Therefore, this study was carried out to identify PKM2 inhibitors from natural products for cancer treatment. Based on the objectives of this study, firstly, plant extract library was established. In order to purify protein for the establishment of enzymatic assay system, pET-28a-HmPKM2 plasmid was transformed to E. coli BL21 (DE3) cells for protein expression and purification. After the validation of enzymatic assay system, plant extract library was screened for the identification of inhibitors of PKM2 protein. Out of 51 plant extracts screened, four extracts Mangifera indica (leaf, seed, and bark) and Bombex ceiba bark extracts were found to be inhibitors of PKM2. In the current study, M. indica (leaf, seed, and bark) extracts were further evaluated dose dependently against PKM2. These extracts showed different degrees of concentration-dependent inhibition against PKM2 at 90-360 μg/ml concentrations. We have also investigated the anticancer potential of these extracts against MDA-MB231 cells and generated dose-response curves for the evaluation of IC50 values. M. indica (bark and seed) extracts significantly halted the growth of MDA-MB231 cells with IC50 values of 108 μg/ml and 33 μg/ml, respectively. Literature-based phytochemical analysis of M. indica was carried out, and M. indica-derived 94 compounds were docked against three binding sites of PKM2 for the identification of PKM2 inhibitors. The results of in silico based screening have unveiled various PKM2 modulators; however, further studies are recommended to validate their PKM2 inhibitory potential via in vitro biochemical assay. The results of this study provide novel findings for possible mechanism of action of M. indica (bark and seed) extracts against TNBC via PKM2 inhibition suggesting that M. indica might be of therapeutic interest for the treatment of TNBC.