Structure-activity Relationship of Sintenin and its Analogues on Six Human Tumor Cell Lines

Cytotoxic compounds are crucial in the course of finding new anti-tumor leading compounds. Chen and his co-workers recently reported the isolation of sintenin 1 which possessed selective cytotoxicity against P-388 cells with an ED50 value of 0.21 μg/mL1. As to our knowledge, this kind of esters should have broader-spectrum of biological activity2,3. Furthermore, the structure of 1 is relatively similar to nelumol B-D, the sinapyl alcohol derivatives isolated from Ligularia nelumbifolia, wh…     

Effect of α-Ketoglutarate on Monoclonal Antibody Production of Hybridoma Cell Lines in Serum-Free and Serum-Containing Medium

Process development and optimization for increase population growth and protein productivity in mammalian cell culture have been studied for many years. In this study, the behavior of hybridoma cells was investigated using six-well micro-titer plate systems with a working volume of 4 ml. Mouse hybridoma cell lines D2 and 2C83G2 were seeded in serum-free and serum-containing media and cultured for 8 days. alpha-Ketoglutarate is an integral component of the tricarboxylic acid (TCA) cycle and is produced from glutamine via glutamate. To study its effect on cell growth, metabolism, and monoclonal antibody (mAb) production, 2 mM alpha-ketoglutarate (pH 7.2) was added in both media at the beginning of the cultivation and in another set after 72 h. High cell density was observed in D2 cell culturing in serum-free medium, while 2C83G2 cell line showed high cell density in serum-containing medium. However, both cell lines cultured in serum-free medium gave viability above 70% when grown for 8 days. The supplement of 2 mM alpha-ketoglutarate supported cell growth and mAb production of both hybridoma cell lines in serum-free and serum-containing medium. The addition of alpha-ketoglutarate at the beginning of the batch cultivation gave better result in cell growth and mAb production as compared to alpha-ketoglutarate supplementation after 72 h. However, addition after 72 h was better than no addition at all. This indicates that alpha-ketoglutarate have a positive effect on production and release of antibody.     

Expression Levels of RFP in Normal and Cancer Human Tissues via Real-time RT-PCR Detection

IntroductionRet finger protein(RFP) was first identified as apart of the human recombined transforming gene, ret,and was a member of the tripartite motif(TRIM) pro-tein family. Itwas postulated thatmultimeric complexesmade of TRIM proteins defined differe…     

Trans-(−)-Kusunokinin: A Potential Anticancer Lignan Compound against HER2 in Breast Cancer Cell Lines?

Trans-(−)-kusunokinin, an anticancer compound, binds CSF1R with low affinity in breast cancer cells. Therefore, finding an additional possible target of trans-(−)-kusunokinin remains of importance for further development. Here, a computational study was completed followed by indirect proof of specific target proteins using small interfering RNA (siRNA). Ten proteins in breast cancer were selected for molecular docking and molecular dynamics simulation. A preferred active form in racemic trans-(±)-kusunokinin was trans-(−)-kusunokinin, which had stronger binding energy on HER2 trans-(+)-kusunokinin; however, it was weaker than the designed HER inhibitors (03Q and neratinib). Predictively, trans-(−)-kusunokinin bound HER2 similarly to a reversible HER2 inhibitor. We then verified the action of (±)-kusunokinin compared with neratinibon breast cancer cells (MCF-7). (±)-Kusunokinin exhibited less cytotoxicity on normal L-929 and MCF-7 than neratinib. (±)-Kusunokinin and neratinib had stronger inhibited cell proliferation than siRNA-HER2. Moreover, (±)-kusunokinin decreased Ras, ERK, CyclinB1, CyclinD and CDK1. Meanwhile, neratinib downregulated HER, MEK1, ERK, c-Myc, CyclinB1, CyclinD and CDK1. Knocking down HER2 downregulated only HER2. siRNA-HER2 combination with (±)-kusunokinin suppressed HER2, c-Myc, CyclinB1, CyclinD and CDK1. On the other hand, siRNA-HER2 combination with neratinib increased HER2, MEK1, ERK, c-Myc, CyclinB1, CyclinD and CDK1 to normal levels. We conclude that trans-(±)-kusunokinin may bind HER2 with low affinity and had a different action from neratinib.     

Differential Activation of Signaling Pathways by UVA and UVB Radiation in Normal Human Epidermal Keratinocytes(†)

Ultraviolet (UV) radiation from the solar spectrum is a major etiological factor for many cutaneous pathologies including cancer. By understanding changes in cell signaling pathways induced by UVA and UVB, novel strategies for prevention and treatment of UV‐related pathologies could be developed. However, much of the information in the literature from various laboratories cannot cross talk because of difficulties associated with the use of ill‐defined light sources and physiologically irrelevant light dosimetry. Herein, we have assessed the effect of exposure of normal human epidermal keratinocytes (NHEK) to UVA (2 and 4 J cm^?2) or UVB (20 and 40 mJ cm^?2) radiation. Employing western blot analysis, we found that exposure of NHEK to UVB, but not UVA, phosphorylates JNK1/2 at Th¹⁸³/Tyr¹⁸⁵, STAT3 at Ser⁷²⁷, AKT at Ser⁴⁷³ and increases c‐Fos expression, whereas exposure to UVA, but not UVB, phosphorylates AKT at Thr³⁰⁸. UVB as well as UVA exposure leads to increased phosphorylation of (1) ERK1/2 at Th²⁰²/Tyr²⁰⁴; (2) p38 at Th¹⁸⁰/Tyr²⁰⁴; (3) STAT3 at Tyr⁷⁰⁵; (4) mTOR at Thr²⁴⁴⁸; and (v) p70S6k at Thr⁴²¹/Ser⁴²⁴; enhanced expression of PI3K (p85) and c‐jun; and nuclear translocation of NFκB proteins. These findings could be considered as a beginning for understanding the differential effects of UVA and UVB in the human skin and may have implications both with respect to risk assessment from exposure to solar UV radiation, and to target interventions against signaling events mediated by UVA and UVB.     

Bioelectrical Impedance Assay to Monitor Changes in Aspirin‐Treated Human Colon Cancer HT‐29 Cell Shape during Apoptosis

Abstract

Physiological cell death occurs primarily through an evolutionarily conserved form of cell suicide termed apoptosis, which may be triggered by cytokines, depletion of growth factors, or certain chemicals. It is morphologically characterized by severe alterations in cell shape like cell shrinkage and disintegration of cell–cell contacts. Aspirin and other nonsteroidal anti‐inflammatory drugs (NSAIDs) inhibit proliferation of human colon cancer cells in vitro. We applied a noninvasive bioelectrical impedance assay referred to as electric cell–substrate impedance sensing (ECIS) in order to monitor the apoptosis‐induced changes in human colon cancer HT‐29 cell shape in an integral and quantitative fashion with a time resolution on the order of minutes. In whole‐cell biosensors the cells are grown directly on the surface of small gold‐film electrodes. From readings of the electrical impedance of the cell‐covered electrode, we deduced alterations of aspirin on HT‐29 cells in cell–cell and cell–substrate contacts. And the apoptosis was verified by transmission electron microscope.     

Phorbol Esters from Jatropha Meal Triggered Apoptosis, Activated PKC-δ, Caspase-3 Proteins and Down-Regulated the Proto-Oncogenes in MCF-7 and HeLa Cancer Cell Lines

Jatropha meal produced from the kernel of Jatropha curcas Linn. grown in Malaysia contains phorbol esters (PEs). The potential benefits of PEs present in the meal as anticancer agent are still not well understood. Hence, this study was conducted to evaluate the cytotoxic effects and mode of actions of PEs isolated from Jatropha meal against breast (MCF-7) and cervical (HeLa) cancer cell lines. Isolated PEs inhibited cells proliferation in a dose-dependent manner of both MCF-7 and HeLa cell lines with the IC₅₀ of 128.6 ± 2.51 and 133.0 ± 1.96 μg PMA equivalents/mL respectively, while the values for the phorbol 12-myristate 13-acetate (PMA) as positive control were 114.7 ± 1.73 and 119.6 ± 3.73 μg/mL, respectively. Microscopic examination showed significant morphological changes that resemble apoptosis in both cell lines when treated with PEs and PMA at IC₅₀ concentration after 24 h. Flow cytometry analysis and DNA fragmentation results confirmed the apoptosis induction of PEs and PMA in both cell lines. The PEs isolated from Jatropha meal activated the PKC-δ and down-regulated the proto-oncogenes (c-Myc, c-Fos and c-Jun). These changes probably led to the activation of Caspase-3 protein and apoptosis cell death occurred in MCF-7 and HeLa cell lines upon 24 h treatment with PEs and PMA. Phorbol esters of Jatropha meal were found to be promising as an alternative to replace the chemotherapeutic drugs for cancer therapy.     

Proteomic Analysis of Differential Protein Expression of Achilles Tendon in a Rabbit Model by Two-Dimensional Polyacrylamide Gel Electrophoresis at 21 Days Postoperation

Postoperative early kinesitherapy has been advocated as an optimal method for treating Achilles tendon rupture. However, an insight into the rationale of how early kinesitherapy contributes to healing of Achilles tendon remains to be achieved, and research in the area of proteomic analysis of Achilles tendon has so far been lacking. Forty-two rabbits were randomized into control group, immobilization group, and early motion group, and received postoperative cast immobilization and early motion treatments. Achilles tendon samples were prepared 21 days following microsurgery, and the proteins were separated with two-dimensional polyacrylamide gel electrophoresis. Differentially expressed proteins were first recognized by PDQuest software, and then identified using peptide mass fingerprinting, tandem mass spectrometry, and database searching. A total of 463 ± 12, 511 ± 39, and 513 ± 80 protein spots were successfully detected in the two-dimensional polyacrylamide gels for the Achilles tendon samples of rabbits in the control group, immobilization group, and early motion group, respectively. There were 15, 8, and 9 unique proteins in these three groups, respectively, and some differentially expressed proteins were also identified in each group. It was indicated that some of the differentially expressed proteins were involved in various metabolism pathways and may play an important role in healing of Achilles tendon rupture. Postoperative early kinesitherapy resulted in differentially expressed proteins in ruptured Achilles tendon compared with those treated with postoperative cast immobilization. These differentially expressed proteins may contribute to healing of Achilles tendon rupture through a mechanobiological mechanism due to the application of postoperative early kinesitherapy.     

LC Determination of Malondialdehyde Concentrations in the Human Umbilical Vein Endothelial Cell Culture Medium: Application to the Antioxidant Effect of Vitexin-2″-O-rhamnoside

Vitexin-2″-O-rhamnoside (VOR) presents the leaves of Crataegus pinnatifida Bge. var. major which plays a role in preventing human pathologies related to oxidative stress, but as the principal component in the leaves, little attention has been devoted to its study of antioxidation of VOR. A simple, rapid and sensitive high-performance liquid chromatography method was developed to determine malondialdehyde (MDA) in ECV304 cell culture medium induced by tert-butyl-hydroperoxide (TBHP). The preparation of analyzed samples involved a one-step derivatization with thiobarbituric acid (TBA). The separation was achieved using the Synergi Hydro-RP, a polar endcapped C₁₈ column (250 × 4.6 mm, 4 μm pore size) with a linear gradient elution of acetonitrile and 10 mmol L^?1 of ammonium acetate aqueous solution (pH 6.8). The calibration curve was linear over the ranges 0.0125–1.25 μmol L^?1 MDA (r = 0.9962). The lower limit of quantification of MDA was 0.0125 μmol L^?1. Relative standard deviations of intra-day and inter-day precision were less than 5.2 and 4.4%. The method with high recovery (95.4 ± 1.3%) was successfully applied to the investigation of antioxidation of VOR by determining MDA in ECV304 cell culture medium.     

Ratio Design of Docetaxel/Quercetin Co-Loading-to-Nanocarrier: Synthesis of PCL–PEG–PCL Copolymer,Study of Drug Release Kinetic and Growth Inhibition of Human Breast Cancer (MCF-7) Cell Line

Russian Journal of Applied Chemistry - The present study aimed to design PCL–PEG–PCL copolymer as novel nanocarrier for co-loading of docetaxel (DTX) and quercetin (Qu) drugs and...     

Redox Mediated Synthesis of Ag-CuO Hybrid Nanoparticles – DNA/BSA Binding Studies and in vitro Evaluation of Anti-cancer Activity on MCF-7 Cancer Cell Line

We herein report a nanoparticle-directed therapeutic approach to breast cancer using Ag-CuO hybrid nanoparticles. The nanoparticles were synthesized through redox-mediated process involving reduction of silver ion on the surface of Cu₂O nanostructures. Structural, microstructural and optical characterization were carried out and the hybrid nanoparticles were found to be phase pure with crystallite size between 100–200 nm and the absorption due to surface plasmon resonance from silver was observed around 460 nm. The binding affinities of the hybrid nanoparticles with the plasma protein BSA and calf thymus DNA were studied and the respective binding constants were found to be 5.1 × 10⁴ and 1.12 × 10⁵ M⁻¹ showing appreciable binding affinity of the substrate. The cytotoxicity of the as-synthesized hybrid nanoparticles induced in breast cancer cells was evaluated in vitro and the obtained results demonstrates the potential anti-cancer activity of the Ag-CuO hybrid nanoparticles against the MCF-7 breast cancer cell line.     

Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA—derivatization and Electrochemical Detection

A novel method for determination of amino acids in individual red blood cells has been developed. In this method, the derivatization reagents (NDA and CN^-) are introduced into living cells by electroporation. After completion of derivatization,the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.     

Two Coordination Modes of Bidentate Aminopyrazine Ligands in Cubane-type Cluster Complex Re4Te4Cl8(C4N3H4)4 · 2DMF

Abstract  A new cubane-type cluster complex Re₄Te₄Cl₄(C₄H₄N₃)₄ · 2DMF has been synthesized by reaction of Re₄Te₄Cl₈(TeCl₂)₄ with 2-aminopyrazine C₄H₅N₃ in DMF. The crystal structure of compound has been solved by X-ray single crystal diffraction method. Crystal data for Re₄Te₄Cl₄(C₄N₃H₄)₄ · 2DMF: a = 22.8718(16) ?, b = 8.5936(7) ?, c = 20.5720(17) ?, β ^o = 106.493(2), V = 3877.1(5) ?³, R ₁ = 0.0466, R _w(F ²) = 0.1191. In the complex bidentate aminopyrazine ligands are coordinated in two different types, namely, two of four aminopyrazine ligands bind to a single rhenium atom, and each of two other ligands is coordinated as bridge between two rhenium atoms. Graphical Abstract  A new cubane-type cluster complex Re₄Te₄Cl₄(C₄H₄N₃)₄  · 2DMF with two coordination modes of bidentate aminopyrazine ligands has been synthesized and structurally characterized.      

A multivariate insight into the in vitro antitumour screen database of the National Cancer Institute: classification of compounds, similarities among cell lines and the influence of molecular targets

A multivariate insight into the in vitro antitumour screen database of the NCI by means of the SIMCA package allows to propose hypotheses on the mechanism of action of novel anticancer compounds. As an example, the application of multivariate analysis to the NCI standard database provided clues to the classification of drugs whose mechanism is either unknown or controversial. Moreover, the influence of intrinsic biochemical cell line properties (molecular targets) on the sensitivity to drug treatment could be evaluated simultaneously for classes of compounds which act by the same mechanism. Interestingly, the present approach can also provide a correlation between the molecular targets and the therapeutical fingerprint of novel active compounds thus suggesting specific biochemical studies for the investigation of new mechanisms of drug action and resistance. The statistical approach reported here represents a valuable tool for handling theenormous data sets deriving from recent genome-wide investigations of gene expression in the NCI cell lines.     

Twelve Million Resolving Power on 4.7 T Fourier Transform Ion Cyclotron Resonance Instrument with Dynamically Harmonized Cell—Observation of Fine Structure in Peptide Mass Spectra

Resolving power of about 12,000 000 at m/z 675 has been achieved on low field homogeneity 4.7 T magnet using a dynamically harmonized Fourier transform ion cyclotron resonance (FT ICR) cell. Mass spectra of the fine structure of the isotopic distribution of a peptide were obtained and strong discrimination of small intensity peaks was observed in case of resonance excitation of the ions of the whole isotopic cluster to the same cyclotron radius. The absence of some peaks from the mass spectra of the fine structure was explained basing on results of computer simulations showing strong ion cloud interactions, which cause the coalescence of peaks with m/z close to that of the highest magnitude peak. The way to prevent peak discrimination is to excite ion clouds of different m/z to different cyclotron radii, which was demonstrated and investigated both experimentally and by computer simulations.