Normal L02 Cells Research Articles

Nanobiofunctionalized chitosan nanocomposite hydrogel as a highly biocompatible cancer drug carrier

AbstractA bio‐inspired drug carrier was developed by dual functionalization of chitosan using L‐glutamic acid (GA) and phyto‐synthesized zinc oxide nanoparticles (ZNPs). A highly porous, three‐dimensional network of nanocomposite hydrogel (GA‐CHGZ) was obtained upon cross‐linking chitosan using biomass‐derived dialdehyde cellulose. The hydrogel was optimally loaded with naringenin (NRG) and further characterized using nuclear magnetic resonance (NMR), Fourier‐transform infrared spectroscopy (FTIR), X‐ray diffraction (XRD), scanning electron microscopy (SEM), and swelling studies. An enhanced NRG loading efficiency of 85.23% was obtained using functionalized hydrogel compared to 52.54% using non‐functionalized hydrogel. Delivery studies displayed a maximum release of 69.63% for 1.0 mg/ml of initial NRG concentration at pH 5, which is a highly preferred condition for cancer therapeutics. While ZNPs’ embedment was instrumental in improving the NRG loading and delivery rates, the GA conjugation increased the stability of NRG in the GA‐CHGZ, aiding sustained NRG release, which followed a non‐Fickian diffusion mechanism with polymer swelling. Antimicrobial potential was explored against Staphylococcus aureus and Trichophyton rubrum strains. The biocompatibility assay using L929 normal cells showed enhanced cell proliferation characteristics for the materials, revealing significant cell viability. The anticancer activity of NRG tested against A431 human skin carcinoma cells increased up to nine‐fold with a reduced IC50 value when a functionalized hydrogel was used instead of pure NRG without the nanocomposite carrier. Thus, the bio‐functionalized drug–carrier system has a promising application for wound healing and topical skin cancer therapies.

Novel acetic acid derivatives containing quinazolin-4(3H)-one ring: Synthesis, in vitro, and in silico evaluation of potent aldose reductase inhibitors.

Aldose reductase (AR) is a crucial enzyme of the polyol pathway through which glucose is metabolized under conditions of hyperglycemia related to diabetes. A series of novel acetic acid derivatives containing quinazolin-4(3H)-one ring (1-22) was synthesized and tested for in vitro ARinhibitory effect. All the target compounds exhibited nanomolar activity against the target enzyme, and all compounds displayed higher activity as compared to the reference drug epalrestat. Among them, Compound 19, named 2-(4-[(2-[(4-methylpiperazin-1-yl)methyl]-4-oxoquinazolin-3(4H)-ylimino)methyl]phenoxy)acetic acid, displayed the strongest inhibitory effect with a KI value of 61.20 ± 10.18 nM. Additionally, these compounds were investigated for activity against L929, nontumoral fibroblast cells, and MCF-7, breast cancer cells using the MTT assay. Compounds 16 and 19 showed lower toxicity against the normal L929 cells. The synthesized compounds' (1-22) absorption, distribution, metabolism, and excretionproperties were also evaluated. Molecular docking simulations were used to look into the possible binding mechanisms of these inhibitors against AR.

Bioorthogonal activation of prodrugs, for the potential treatment of breast cancer, using the Staudinger reaction.

Selective prodrug activation at a tumor site is crucial to maximise the efficiency of chemotherapy approaches and minimise side effects due to off-site activation. In this paper, a new prodrug activation strategy is reported based on the bioorthogonal Staudinger reaction. The feasibility of this prodrug activation strategy was initially demonstrated using 9-azido sialic acid 4 as a trigger and two novel triphenylphosphine-modified N-mustard-PRO 10 and doxorubicin-PRO 12 prodrugs in an HPLC-monitored release study. Then, the azide reporter group was introduced on cancer cells' surfaces through metabolic glycoengineering of sialic acid-rich surface glycans using azide-modified monosaccharides (9-azido sialic acid 4, tetra-O-acetylated-9-azido sialic acid 5 and tetra-O-acetyl azidomannosamine). Next, the N-mustard-PRO 10 and doxorubicin-PRO 12 prodrugs were employed in vitro with the bioengineered cells, and activation of the prodrugs, which allowed selective release of the cytotoxic moiety at the tumour cell, was assessed. Release of the parent drugs from the prodrugs was shown to be dependent on the level of metabolic labelling, where tetra-O-acetyl azidomannosamine allowed the highest level of azide reporter generation in tumor cells and led to full recovery of the parent cytotoxic drug's potency. The selectivity of azide expression on breast cancer MCF-7 cells versus normal fibroblast L929 cells was also probed, with the 9-azido sialic acid and tetra-O-acetylated-9-azido sialic acid showing ∼17-fold higher azide expression on the former. Taken together, these data demonstrate the feasibility of the Staudinger reaction for selective activation of prodrugs targeted to the MCF-7 breast cancer cells.

Design and Synthesis of Imidazole Derivatives as Anticancer Agents and Potential Aromatase Inhibitors

In this study, imidazole derivative compounds were synthesized using the Debus-Radziszewski method. The chemical structures of the compounds were characterized by spectroscopic methods. The effects of target compounds on MCF7 (CRL-3435) were examined and their IC50 values and percent viability were calculated. In addition, the cytotoxic effects on the L929 (CCL-1) normal cell line were evaluated in order to determine the selectivities of the compounds. Then, the inhibition values of aromatase enzyme of the compounds were calculated and compared to the reference compound. When the results were examined, it was observed that Compound la caused the death of breast cancer cells, although not as much as cisplatin, but did not harm healthy cells. In this respect, it was determined that compound Ia has a promising anticancer effect as an aromatase inhibitor.

Biological Activities of Essential Oil of Vitex altissima Leaves and Inhibition Potential towards Phosphoinositide-3 Kinase (PI3K) Enzyme by Molecular Docking

This study aims to employ molecular docking to verify the efficacy of a few bioactive compounds of pharmacological significance extracted from Vitex altissima. GC/MS and GC/FID were used for chemical profiling. An in vitro study for anticancer potential done using MTT assay with DLD-1 cancer cell line and L929 normal cell line. All the bioactive compounds were docked against phosphoinositide-3 kinase (PI3K) using AutoDock 4.2.6 software. The identified 22 compounds (89.91%) with allo-aromadendrene (29.82%), E-phytol (16.08%) α-humulene (14.04%), β-caryophyllene (4.18%), α-santalol (3.64%) and spanthulenol (2.23%) as the dominant sesquiterpenoid compounds. The study indicates effective anticancer and antioxidant potential with the oil. Apoptosis evaluation revealed that the cells experience necrosis after treatment with Vitex altissima leaf oil. The deregulation of enzyme PI3K is liable for various types of cancers. The ADMET properties of respective compounds reveal their pharmacokinetic properties. All compounds subjected to docking satisfied Lipinski’s rule of five, indicating the scope for their use as oral drugs.

New2-((2-(2,4-dinitrophenyl)hydrazineeylidene) derivatives: design, synthesis, in silico, and in vitro anticancer studies

A series of novel hydrazone compounds have been synthesized by the condensation of hydrazines and different substituted salicylaldehydes at a molar ratio of 1:1 in one step reaction and characterized by FT-IR, ESI-MS, 1H NMR, and single crystal x-ray diffraction. The crystal structure of the compound shows a trans configuration around the C = N bond and triclinic system with P −1/-p 1. Synthesized compounds were screened for cytotoxicity activities against A375 (melanoma), HT-29 (Colon), and A549 (lung) cancer cell lines. Among them, compound 2 exhibited the highest cytotoxic effect against the A375 cell line (IC50 = 0.30 µM) and HT-29 cell line (1.68 µM), compared to those of apatinib as a reference standard drug (0.28, 1.49 µM, respectively). The cytocompatibility assay on the L929 normal cell line and the hemolysis assay on human RBC were used to validate the non-toxic action. From DFT calculation, the various parameters such as HOMO-LUMO energies, Hirshfeld, and MEP have been studied. Furthermore, in silico molecular docking with three receptors was studied. Among four compounds, compound 2 has the lowest binding energy against cyclin dependent kinase (ΔGb = −9.3 kcal/mol). In addition to this, molecular dynamics (MD) simulation was also performed. Based on this study, these novel hydrazones can be considered a promising anticancer agent due to their potent cytotoxicity activities and computational analysis. Communicated by Ramaswamy H. Sarma

Fabrication of a novel bioactive chitosan based biocomposite from Opuntia ficus-indica fruit gum and evaluation of anticancer activity in bone MG63 cancer and L929 normal cell lines

In this paper, a novel bioactive Chitosan-Lignin/n-Hydroxyapatite/Opuntia ficus-indica -Fruit gum (Cs-Lg/n-Hap/OFI-Fg) biocomposite was fabricated. The as-obtained biocomposite properties are analyzed by optical and structural investigation. The biological properties like antibacterial, antioxidant, and anticancer ability were evaluated for the biocomposites. The FT-IR spectrum result shows the biocomposites possess significant intermolecular interaction with biomolecules. The powder XRD result indicated the as-obtained Cs-Lg/n-Hap/OFI-Fg biocomposites are crystalline in nature. The microhardness test shows the addition of fruit gum increases the mechanical strength of the biocomposite. The thermal stabilities of the as-obtained Cs-Lg/n-Hap/OFI-Fg biocomposite were higher than to Cs-Lg/n-Hap composite as revealed by TGA analysis. The SEM images show an addition of fruit gum leads to the formation of spherical-shaped discrete particle morphology evidenced by TEM analysis. The antioxidant and antibacterial activity of bioactive biocomposite shows better activity due to the presence of numerous biomolecules. The MTT assay result revealed significant activity in as-obtained Cs-Lg/n-Hap/OFI-Fg biocomposite compared to Cs-Lg/n-Hap composite against MG63 bone cells and L929 cells due to the active biomolecules extant in fruit gum. The anticancer results revealed that 5 µg/ml concentration biocomposite is a significant role against bone cancer cells which proves that the as-obtained composites could be more valuable for the treatment of bone deficiency.

Fabrication of poly(ϵ‐caprolactone)/paclitaxel (core)/chitosan/zein/multi‐walled carbon nanotubes/doxorubicin (shell) nanofibers against MCF‐7 breast cancer

AbstractIn the present study, paclitaxel (PTX), multi‐walled carbon nanotubes (MWCNTs), and doxorubicin (DOX) have been simultaneously doped into the poly(ϵ‐caprolactone) (PCL)/chitosan/zein core‐shell nanofibers to increase its cytotoxicity for MCF‐7 breast cancers killing. The physico‐chemical properties of synthesized nanofibers were determined by scanning electron microscope, Fourier‐transform infrared spectroscopy, tensile strength, and degradation rate determinations. The in vitro release studies demonstrated the sustained release of drugs from core‐shell nanofibrous scaffold. The cytotoxicity and compatibility of core‐shell nanofibers were investigated by their treating with MCF‐7 breast cancer cells and L929 normal cells, respectively. PCL/PTX/chitosan/zein/MWCNTs/DOX core‐shell nanofibers containing 1 wt% MWCNTs, 100 μg ml−1 DOX and 100 μg ml−1 PTX had a high biocompatibility with a 84% MCF‐7 cancer cells killing. The in vivo studies revealed the synergic effects of MWCNTs and anticancer drugs on the tumor inhibition. This method could be considered as a new way for developing of MWCNTs loaded‐nanofibers for cancer treatment in future.

Nanoaggregates of Disulfide-Decorated TrxR Inhibitor Promote Cellular Uptake, Selective Targeting, and Antitumor Efficacy.

Three self-assembled nanoaggregates (CPUL1-LA NAs, CPUL1-DA NAs, and CPUL1-AA NAs) were constructed through lipoic acid (LA), dithiodipropionic acid (DA), and adipic acid (AA) decorated TrxR inhibitor (CPUL1), respectively. Measurements of DLS, TEM, UV-vis, fluorescence, 1H NMR, ITC, and MTT assays verified disulfide-containing CPUL1-LA NAs and CPUL1-DA NAs spontaneously assembled carrier-free nanoparticles in aqueous solution, which possessed high drug contents, excellent stability, improved cytotoxicity against HUH7 hepatoma cells, and potential biosafety because of low cytotoxicity against L02 normal cells. In contrast, disulfide-free CPUL1-AA NAs happened to aggregate and precipitate after 48 h, which showed distinct instability in aqueous solution. Thus, disulfide units seemed to be crucial for constructing controllable and stable nanoaggregates. While measuring the reduction of nanoaggregates by TrxR/NADPH and GSH/GR/NADPH, cyclic disulfide of LA and linear disulfide of DA were verified to endow the nanoaggregates with targeting ability to respond specifically to TrxR over GSH. Furthermore, by tests of flow cytometry, fluorescence images, and CLSM, both CPUL1-LA NAs and CPUL1-DA NAs displayed a faster cellular uptake characteristic to be internalized by cancer cells and could generate more abundant ROS to induce cell apoptosis than that of free CPUL1, resulting in significantly improved antitumor efficacy against HUH7 cells in vitro.

Significant effect on the photo conversion performance of non-stoichiometric tungsten oxide with various reaction solvents

Photothermal therapy (PTT) is a new emerging treatment method in recent years which has the advantages of non-invasive and wide adaptability. The key to PTT is the development of effective photothermal agents. Herein, the non-stoichiometric tungsten oxides were prepared in a facile solvothermal method with various reaction solvents (ethanol, n-propanol, isopropanol) and time. The obtained samples were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscope, photoluminescence and UV–vis–NIR adsorption spectroscopy. The kind of solvent had a significant effect on the phase, microstructure, photo conversion efficiency. Once n-propanol was used as solvent, the samples possessed high crystallinity, uniform fusiform morphology, good dispersibility, obvious absorbance of NIR and the photothermal conversion efficiency of 35.6%. Under the 808 nm near-infrared light irradiation, in vitro cell tests were further conducted on L929 normal and MCF-7 tumor cells for comparative experiments. The outstanding properties make the samples highly potential agents for PTT.

A New 7‐Azaindole Structure Analog: Molecular Docking, Synthesis and Preliminary Biological Activity in Vitro for Anticancer

In this work, a series of 7-azaindole analogs were designed by the bioisosteric principle based on the pharmacodynamic parent nucleus. Moreover, 5-[(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl]-N-{[6-(trifluoromethyl)pyridin-3-yl]methyl}pyrimidin-2-amine (compound P1) with the strongest interaction with colony-stimulating factor 1 receptor (CSF-1R) was screened by molecular docking. Compound P1 was successfully prepared by the six-step reaction with HPLC purity of 99.26 % and characterized by 1 H-NMR and ESI-MS spectra. In vitro bioactivity study showed that compound P1 appeared the cytotoxicity to MCF-7 and A549 cells, especially to HOS cells (IC50 =88.79±8.07 nM), while it had lower toxicity to normal L929 cells (IC50 =140.49±8.03 μM). In addition, compound P1 could induce HOS cell death by apoptosis and blocking the G0/G1 phase at nanomolar concentrations. The obtained results indicated that compound P1 might be a promising candidate compound for anticancer drug.

Bioactivity of crude fucoidan extracted from Sargassum ilicifolium (Turner) C. Agardh

Fucoidan derived from brown algae has been shown to exhibit antitumor and antioxidant effects, so research on sulfated polysaccharides is increasing. The purpose of this study was to evaluate the characteristics and biological activity of fucoidan that was extracted at two temperatures (65 and 80 °C) from Sargassum ilicifolium (Turner) C. Agardh from five regions of Taiwan. The data show that there are significant differences in the yield, sulfate and total sugar content of Sargassum ilicifolium (Turner) C. Agardh grown in different locations in the same sea area. HPLC was used to determine the monosaccharide compositions of the fucoidan, which contains fucose, mannose, mannose, glucose and galactose and have a low molecular weight of less than 5 kDa, and then we will select the algae collected in Fugang, Taitung, for further biological activity research. The sampled Sargassum ilicifolium (Turner) C. Agardh at all five locations has a good polyphenol content, and it shows great DPPH radical scavenging activity, ABTS radical scavenging activity, Ferrous ion-chelating activity and Reducing power. The Sargassum ilicifolium (Turner) C. Agardh that was collected from Taitung Fugang is not toxic to L929 normal cells, but for A549 cancer cells and HCT116 cancer cells, it is known from the results that it has good cytotoxicity for A549 cancer cells. Thus, this study found that the Sargassum ilicifolium (Turner) C. Agardh that was collected from Taitung Fugang has significant antioxidant and anticancer properties.

Thymol has anticancer effects in U-87 human malignant glioblastoma cells.

Thymol (2-isopropyl-5-methylphenol) is a colorless crystalline derivative of cymene, that possesses pleotropic pharmacological properties, including analgesic, antibacterial, antispasmodic, and anti-inflammatory activities. Thymol has also been recognized for its beneficial effect as an anti-tumor agent, but the precise mechanism for this has not been fully elucidated. We aimed to identifying whether thymol exerts anti-cancer activity in human U-87 malignant glioblastoma (GB) cells (U-87). Cell viability and apoptosis was evaluated in U-87 cells treated with thymol at different concentrations. Reactive oxygen species (ROS) production, mRNA expressions of apoptosis-related genes and cell cycle characteristics were assessed. The cytotoxic activity of the co-exposure of thymol and temozolomide (TMZ) was also evaluated. The half-maximal inhibitory concentration (IC50) of thymol in the U-87 cells was 230μM assessed at 24h after exposure. Thymol did not exhibit any cytotoxic effects on normal L929 cells at this concentration. Thymol treatment increased the expression of Bax and p53, and also increased apoptotic cell death, and excessive generation of ROS. Moreover, the cytotoxic activity of thymol on the U-87 cells may be related to the arrest of the cell cycle at the G0/G1 interface. Combination therapy showed that the cytotoxic effects of thymol synergized with TMZ, and combined treatment had more cytotoxic potential compared to either of the agents alone. Our data indicate the potential cytotoxic activities of thymol on U-87 cells. Further studies are required to evaluate the spectrum of the antitumor activity of thymol on GB cells.

Synthesis, characterization and molecular docking studies of phenoxyimine based ligands: Cytotoxicity, hemolytic activity and antioxidant assessment

A series of phenoxyimine compounds were designed and synthesized by the condensation of different substituted salicylaldehyde and aniline at the molar ratio of 1:1 in one step reaction. All synthesized compounds were characterized by analytical and spectroscopic techniques. All compounds were screened for cytotoxicity activities against different cancer cell lines. Compound 3 exhibits significant cytotoxic activity with an IC50 value of 1.099 µM, 1.132 µM against COLO-205 (Colorectal adenocarcinoma) and A-549 (Lung carcinoma) cell line respectively, while compound 4 exhibits noteworthy cytotoxic activity with an IC50 value of 0.52 µM against MDA-MB-231(Breast adenocarcinoma). Similarly, other compounds also exhibited good cytotoxicity properties. Furthermore, the phenoxyimine compounds showed the greatest antioxidant activity against DPPH radicals. The nontoxic activity was confirmed by cytocompatibility assay on L929 normal cell line and hemolysis assay on human red blood cells. Additionally, in silico molecular docking of all compounds were carried out against EGFR, HER-2 and VEGF. Compound 3 having minimum binding energy ΔGb = -8.81 kcal/mol, ΔGb = -10.49 kcal /mol, ΔGb = -9.99 kcal /mol, was bound into the active pocket of EGFR, HER-2 and VEGF respectively.

Chemistry of glycol nucleic acid (GNA): Synthesis, photophysical characterization and insight into the biological activity of phenanthrenyl GNA constituents

The knowledge pertaining to the chemistry and biological activity of glycol nucleic acid (GNA) components, like nucleosides and nucleotides, is still very limited. Herein we report on the preparation of the uracil nucleoside (1) and nucleotide ester GNA (2). The compounds are functionalised with a luminescent phenanthrenyl group. In DMSO, 1 and 2 are brightly fluorescent, with emission maxima at 390 nm, nanosecond decay times (0.6 and 0.75 ns, respectively), and quantum yields of ca. 0.2. In the solid phase, they show excimeric emission, with maxima at 495 nm (1) and 432 nm (2), and decay times of 3.7 ns (1) and 2.9 ns (2). The anticancer activity of the GNA components, as well as gemcitabine hydrochloride, used as a reference drug, were examined in vitro against human cancer HeLa and Ishikawa cells, as well as against normal L929 cells, using a battery of biochemical assays. Furthermore, biodistribution imaging studies were carried out in HeLa cells, with luminescence confocal microscopy, which showed that the compounds localized mainly in the lipophilic cellular compartments. Nucleoside (1) and nucleotide ester (2) features two different anticancer activity profiles. At 24 h of treatment, the nucleoside acts mainly as a toxin and induces necrosis in HeLa cells, whereas the nucleotide ester exhibits pro-apoptotic activity. At longer treatment times (72 h), the nucleoside and the reference, gemcitabine hydrochloride, featured almost identical signs of anticancer activity, such as S-phase cell cycle arrest, proliferation inhibition, and apoptosis induction. In view of this data, one can hypothesize that despite the structural differences, the newly obtained phenanthrenyl GNA nucleoside (1) and gemcitabine may share a common mechanism of anticancer activity in HeLa cancer cells. The GNA components were also examined as antiplasmodial agents against Plasmodium falciparum, in vitro. Nucleoside (1) was found to be more potent than nucleotide (2), displaying activity in the low micromolar range. Furthermore, both phenanthrene derivatives were found to display resistance indices at least 9-fold lower than chloroquine diphosphate (CQDP).

Curcumin Nanoparticles as Promising Anticancer Agent Against Human Colorectal Cancer Cell Lines

Curcumin, the main bioactive material found in turmeric, has been extensively studied as a curative measure for variety of diseases. Besides its high therapeutic value, curcumin possess some inherent limitations like poor aqueous solubility, less bioavailability, rapid metabolic rate, and fast elimination from the body. In the present study, nanoformulation of curcumin was discussed to improve its potentials and to reduce the disadvantages. The curcumin nanoparticles have been synthesized by wet-milling technique and have been characterized using FTIR, UV-Visible spectroscopy, SEM-EDS, TEM, and XRD. The results showed that the particles have a uniform dispersion and possess spherical shape with size ranging from 10-30 nm. FTIR studies showed that there was no change in the chemical structure of curcumin during the nanoparticle synthesis process. The anticancer activity of the synthesized nanoparticles was studied against DLD-1 (colorectal cancer) cell lines and cytotoxic evaluation was done against L929 (fibroblast) cells. It was observed that the curcumin nanoparticles significantly reduce the cell viability of DLD-1 cells in a concentration dependent manner and less toxic on normal L929 cells.

Characterization of a novel squalene-rich oil: Pachira macrocarpa seed oil.

Pachira macrocarpa is a woody oil crop with high economic and ornamental value. Although P. macrocarpa seeds are rich in oil, little information has been reported about its characterization. In this study, the fatty acids, minor components (tocopherols, squalene, phytosterols, and total phenols), antioxidant activity, cytotoxicity, thermal, and rheological behavior of the P. macrocarpa seed oil (PSO) were investigated for the first time. The results showed that the seeds contained 43.34% lipid, which was mainly composed of palmitic acid (49.96%), linoleic acid (31.22%), and oleic acid (13.48%). The contents of tocopherols, squalene, phytosterols, and total phenols in PSO were 42.01mg/100g, 96.78mg/100g, 119.67mg/100g, and 3.79mg GAE/100g, respectively. PSO showed relatively strong DPPH radical scavenging capacity (93.47µmol TE/100g) and high melting point (20.8°C). In addition, the oil exhibited Newtonian flow behavior and was not toxic to normal L929 cells at concentrations of 500-8000µg/ml. As a whole, PSO may be considered as a valuable source for new multipurpose products for industrial utilization. PRACTICAL APPLICATION: Pachira macrocarpa is a woody oil crop and its seeds are rich in oil. Our study has investigated the physicochemical properties and chemical composition of the P. macrocarpa seed oil (PSO). The present study revealed PSO had potential as an edible oil, and it may be considered as a valuable source for new multipurpose products for food industrial utilization.

In vitro and in vivo anti-cancer effects of hibernating common carp (Cyprinus carpio) plasma on metastatic triple-negative breast cancer

Uncontrollable proliferation is a hallmark of cancer cells. Cell proliferation and migration are significantly depressed during hibernation state. Many studies believe some factors in the plasma of hibernating animals cause these effects. This study aimed to assess the anti-cancer effects of hibernating common carp (Cyprinus carpio) plasma on 4T1 cancer cells in vitro and in vivo. The effect of hibernating plasma on cell viability, morphology, migration, apoptosis rate, and cell cycle distribution of 4T1 cells was investigated in vitro and in vivo. Hibernating plasma at a concentration of 16 mg/ml significantly reduced the viability of 4T1 cancer cells, without any toxicity on L929 normal fibroblast cells. It could change the morphology of cancer cells, induced apoptosis and cell cycle arrest at the G2/M phase, and inhibited migration. Furthermore, intratumoral injection of hibernating plasma (200 µl, 16 mg/ml) in the tumor-bearing mice caused a significant inhibition of 4T1 breast tumors volume (46.9%) and weight (58.8%) compared with controls. A significant decrease in the number of metastatic colonies at the lungs (80%) and liver (52.8%) of hibernating plasma-treated animals was detected which increased the survival time (21.9%) compared to the control groups. Immunohistochemical analysis revealed a considerable reduction in the Ki-67-positive cells in the tumor section of the hibernating plasma-treated animals compared with controls. Taken together, the SDS-PAGE and mass spectrometry analysis indicated the alpha-2-macroglobulin level in the hibernating fish plasma was significantly increased. It could exert an anti-cancer effect on breast cancer cells and suggested as a novel cancer treatment strategy.

Ethanol promoting the upregulation of C-X-C Motif Chemokine Ligand 1（CXCL1） and C-X-C Motif Chemokine Ligand 6（CXCL6） in models of early alcoholic liver disease

ABSTRACT Alcoholic liver disease (ALD) denotes a series of liver diseases caused by ethanol. Recently, immune-related genes (IRGs) play increasingly crucial role in diseases. However, it’s unclear the role of IRGs in ALD. Bioinformatic analysis was used to discern the core immune-related differential genes (IRDGs) in the present study. Subsequently, Cell Counting Kit-8 say, oil red O staining, and triglyceride detection were employed to explore optimal experimental conditions of establishing hepatocellular models of early ALD. Ultimately, real-time reverse transcription-PCR and immunohistochemistry/immunocytochemistry methods were adopted to verify the expressions of mRNA and proteins of core IRDGs, respectively. C-X-C Motif Chemokine Ligand 1 (Cxcl1) and Cxcl6 were regarded as core IRDGs via integrated bioinformatics analysis. Besides, Lieber Decarli Ethanol feeding and 200 mM and 300 mM ethanol stimulating L02 cells for 36 h can both successfully hepatocellular model. In ethanol groups, the levels of CXCL1 and CXCL6 mRNA were significantly upregulated than pair-fed groups (P < 0.0001). Also, immunohistochemistry revealed that positive particles of CXCL1 and CXCL6 in mice model of early ALD were obviously more than control groups (P < 0.0001). Besides, in L02 hepatocytes stimulated by ethanol, CXCL1 and CXCL6 mRNA were over-expressed, compared with normal L02 cells (P < 0.0001). Meanwhile, immunocytochemistry indicated that CXCL1 and CXCL6 proteins in hepatocellular model of early ALD were higher than normal L02 hepatocytes stimulus (P < 0.0001). Ethanol promoted the upregulation of Cxcl1 and Cxcl6 mRNA and proteins in models of early ALD, denoting their potentiality of acting as biomarkers of ALD.

Sodium metabisulfite in dried plum and its cytotoxic effects on K-562 and L-929 normal cell lines.

In the present study, the concentration of sodium metabisulfite (SMB) in dried plums and its toxicity effects on the cell lines of K-562 (human leukemia cell line) and L-929 (normal fibroblast cell line) were measured. Samples of dried plums were randomly collected from the shops located in Neyshabur and Mashhad (Iran). SMB residue was measured using iodometric titration and high-performance liquid chromatography. To analyze the cytotoxicity, the cells were treated with various concentrations of SMB, and cell viability was determined by the MTT and LDH methods. The average concentration of SMB in the samples of dried plums was selected to evaluate the apoptosis/necrosis by flow cytometer. The expression analysis of apoptosis marker genes (BAX, Bcl-2, and P53) was also assessed. Results indicated that the average concentration of SMB residue in 12 samples of dried plum was 516 ± 285.39mg/kg. When K-562 cells were treated with 500mg/L of SMB, apoptosis increased significantly (p<0.01). The IC50 of SMB for K-562 and L-929 cells after a 48-h exposure was 200.31 and 257.82mg/L, respectively. SMB-treated cells showed that cell viability in both cell lines decreased in a dose-dependent manner after 72h (p<0.01). The percentage of apoptotic but not necrotic cells was 69.49% for K-562 and 77.32% for L-929 cells, whereas apoptosis of untreated control cells was 0.17%. Our findings also showed an opposite mRNA expression of Bcl-2 (anti-apoptotic marker) and Bax2 (pro-apoptotic marker) when k-562 cells were treated with SMB. The results indicated that the concentration of sulfite residue in some dried plums poses a cell toxicity risk for normal cells. PRACTICAL APPLICATION: The results of current study provide important information concerning the toxicological effects of SMB, and give a warning that it needs to be replaced by natural products for fruit drying processes.