Human Cervical Adenocarcinoma Cells Research Articles

Chemical composition and cytotoxic activity of the essential oils of Cymbopogon citratus L. Grown in phu tho province

Culms and leaves of Cymbopogon citratus L. were collected from two regions of Phu Tho province (Thanh Son and Phu Ninh) and used as materials for essential oil extraction. Oils obtained were steam-distilled, analyzed for chemical composition and evaluated for cytotoxic activity against three different cancer cell lines. The GC/MS analysis showed that citral is the major content of the steam-distilled essential oils which was found in the range of 64.15-76.22%. Camphene was found only in culm oils of both regions but it was not detected in the leaf oils. Interestingly, the isomer forms of ocimene present at higher content in the culm oils than in the leaf oils whereas myrcene content in the leaf oils is higher than that in the culm oils. In a cytotoxicity test, four essential oils of culms and leaves of C. citratus from Thanh Son and Phu Ninh showed potent activity against A549 (human lung carcinoma) cell line with the IC50 values ranging from 4.01±0.39 to 6.3±0.54 µg/ml. The essential oils (culms and leaves) from Phu Ninh exhibited moderate effects on the Hela (human cervical adenocarcinoma) cells with the IC50 values of 19.43±1.16 and 42±2.41 µg/ml, respectively. However, they were inactive against the human hepatocellular carcinoma Hep3B cell line. The essential oils from Thanh Son exhibited potent cytotoxic activity against Hela and Hep3B cell lines with the IC50 values ranging from 1.18±0.26 to 8.91±0.32 µg/ml. The results indicated that the essential oils of C. citratus from Thanh Son, Phu Tho could be considered as a promising candidate for the natural sources of anticancer agents.

High-Resolution Adhesion Kinetics of EGCG-Exposed Tumor Cells on Biomimetic Interfaces: Comparative Monitoring of Cell Viability Using Label-Free Biosensor and Classic End-Point Assays.

A high-throughput label-free resonant waveguide grating biosensor, the Epic BenchTop, was utilized to in situ monitor the adhesion process of cancer cells on Arg-Gly-Asp tripeptide displaying biomimetic polymer surfaces. Using highly adherent human cervical adenocarcinoma (HeLa) cells as a model system, cell adhesion kinetic data with outstanding temporal resolution were obtained. We found that pre-exposing the cells to various concentrations of the main extract of green tea, the (−)-epigallocatechin gallate (EGCG), largely affected the temporal evolution of the adhesion process. For unexposed and low dosed cells, sigmoid shaped spreading kinetics was recorded. Higher dose of EGCG resulted in a complete absence of the sigmoidal character, and displayed adsorption-like kinetics. By using the first derivatives of the kinetic curves, a simple model was developed to quantify the sigmoidal character and the transition from sigmoidal to adsorption-like kinetics. The calculations showed that the transition happened at EGCG concentration of around 60 μg/mL. Using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide end-point assay, we concluded that EGCG is cytostatic but not cytotoxic. The effect of EGCG was also characterized by flow cytometry. We concluded that, using the introduced label-free methodology, the shape of the cell adhesion kinetic curves can be used to quantify in vitro cell viability in a fast, cost-effective, and highly sensitive manner.

Selection and identification of novel peptides specifically targeting human cervical cancer.

Cervical cancer is the second most commonly diagnosed cancer and the third leading cause of cancer deaths among females in underdeveloped countries. This study aimed to identify several novel cervical cancer-specific targeting peptides (CSPs) to provide new methods for the effective diagnosis and treatment of cervical cancer. Peptide library screening in vivo was performed on human cervical cancer xenografts with Ph.D.™-12 and C7C phage display peptide libraries. Two specific peptide sequences (GDALFSVPLEVY and KQNLAEG), which were enriched in tumors, were screened, and respectively, named CSP-GD and CSP-KQ through three rounds of biopanning. The in vivo tumor-targeting ability of these peptides was identified by injecting them into mice with cervical cancer xenograft. CSPs were compounded and labeled with fluorescein isothiocyanate (FITC). The specificity and affinity of FITC-CSPs were evaluated in human cervical cancer cell lines and tissue microarrays in vitro by immunofluorescent staining. Results showed that FITC-CSP-GD and FITC-CSP-KQ evidently and specifically bound to the cell membrane and cytoplasm of SiHa, ME-180, and C-33A cells in vitro. In human cervical cancer tissue, FITC-CSP-GD and FITC-CSP-KQ strongly targeted human cervical adenocarcinoma and cervical squamous cell carcinoma tissues, respectively. A bright FITC signal was located mainly on the cell membrane and cytoplasm of tumor cells. In conclusion, the novel 12-residue peptide CSP-GD and 7-residue peptide CSP-KQ could specifically target human cervical cancer and may have the potential to be used in the diagnosis and targeted therapy of cervical cancer.

Cytotoxicity, genotoxicity and uptake detection of folic acid-functionalized green upconversion nanoparticles Y2O3/Er3+, Yb3+ as biolabels for cancer cells

Upconversion nanoparticles (UCNPs) have been used as biolabels for cancer cells due to their ability to absorb near-infrared photons and upconvert them into visible radiation. We reported the synthesis of UCNPs Y2O3/Yb3+, Er3+ (1, 1 mol%), which upon excitation with infrared photons (λ = 980 nm) emit green color with a maximum peak centered at λ = 550 nm. UCNPs were functionalized with folic acid (UCNPs-NH2-FA) and analyzed by transmission electron microscopy, Fourier transform infrared spectroscopy, XRD, DLS and photoluminescence measurements. UCNPs-NH2-FA had a particle size of 70 ± 10 nm and exhibit a good luminescence spectrum in comparison with bare UCNPs. Cytotoxicity of different concentrations of bare and functionalized UCNPs was measured with the MTT assay in three cancer cell lines: human cervical adenocarcinoma (HeLa) and human breast adenocarcinoma cells (MDA-MB-231 and MCF-7). Some concentrations of bare UCNPs were cytotoxic for cells; however, after been functionalized, UCNPs resulted to be non-cytotoxic. Genotoxicity of bare and functionalized UCNPs was performed by the comet assay, and no DNA damage was found for any concentration. The internalization of UCNPs-NH2-FA into cancer cells was confirmed by confocal microscopy showing a cytoplasmic fluorescence signal. UCNPs-NH2-FA were used to detect cancer cells in suspension by flow cytometry, with a specific green fluorescent signal for effective detection of cells. These results confirm that functionalized UCNPs can be used without any cytotoxic or genotoxic effects for bioimaging to detect and visualize cancer cells.

Selective Cell Isolation by Transferrin Functionalized Silane–Carbon Soot Mediated Superhydrophobic Micropatterns

AbstractSurfaces that facilitate selective cell adhesion using specific targeting moieties have great implications in diagnostics, tissue engineering, and high throughput screenings. However, designing robust and spatially confined micropatterns for selective cell isolation on portable platform is highly challenging. Here, wettable, silane (Si) micropatterns holding covalently attached transferrin (Tf) for targeting Tf overexpressing cancer cells are reported. These micropatterns are separated by carbon soot based superhydrophobic regions that turn these targeting sites into surface tension confined “microwells.” These microwells facilitate capture of human colorectal carcinoma cells (HCT 116) and human cervical adenocarcinoma cells (HeLa) by confining their attachment to wettable region, thereby making isolation and spotting of the targeted cells more efficient. In addition, owing to its transparent trait, the Tf conjugated wettability based patterned chip offer real time optical monitoring of cell adhesion, cell growth, and cell behavior. The specific cell isolation using such surface has applications in devising cancer recurrence monitoring tests.

Identification of cytotoxic metabolites from Mahonia aquifolium using 1H NMR-based metabolomics approach

Herein, we propose a 1H NMR-based metabolomics method to reveal cytotoxic metabolites from Mahonia aquifolium stem-bark. Primary and secondary metabolites in the Mahonia aquifolium extracts were identified by thorough analysis of 1H and 2D NMR spectra, without prior isolation. An OPLS multivariate analysis method was used to correlate the chemical composition of the plant extracts with the results of cytotoxic activity against Human cervical adenocarcinoma cell line. Protoberberine alkaloids berberinе and palmatine, along with bisbenzylisoquinoline alkaloid berbamine were identified as the most influential in the OPLS model, with the highest cytotoxic activity.

Synthesis and Photodynamic Activity of 5,10,15-Tris(p-chlorophenyl)-20-(2- hydroxy-3-methoxyphenyl)-21H,23H-porphyrin

Photodynamic therapy (PDT) is an effective option for the treatment of solid neoplastic tumors, as well as non-malignant diseases such as dermatological, ophthalmological and benign pathologies. PDT destroys cancer cells by photochemical generation of reactive oxygen species upon absorption of visible light by a photosensitizing agent (PS). Here, we describe the synthesis and characterization of the unsymmetrical A3B porphyrin 5,10,15-Tris(p-chlorophenyl)-20-(2-hydroxy-3-methoxyphenyl)-21H,23H-porphyrin, which presents different groups at the meso positions, p-chlorophenyl and 2-hydroxy-3-methoxyphenyl and is a novel target molecule in order to evaluate the synergistic effect of these hydrophobic and hydrophilic moieties distributed unsymmetrically on the porphyrin macrocycle. We demonstrated the in vitro cytotoxic effect of the synthesized porphyrin, by the WST-1 assay on a cultured human cervical adenocarcinoma cell line (HeLa ATCC: CCL-2), as well as the presence of apoptosis by T.U.N.E.L. assay. We compared the phototoxicity and intrinsic cytotoxicity of the PS and found a considerable difference in the IC50 between phototoxic activity (1.5 μM) and intrinsic cytotoxicity (7 μM). The results of the induction of apoptosis on HeLa cells treated with the unsymmetrical porphyrin at a final concentration of 80 μM showed 15% apoptotic cells, whereas the untreated HeLa cells had only 2.5% apoptotic cells.

Growth inhibitory effect of the Src inhibitor dasatinib in combination with anticancer agents on uterine cervical adenocarcinoma cells.

Uterine cervical adenocarcinoma has a poor clinical prognosis when compared with squamous cell carcinoma. Therefore, the development of new treatment strategies for uterine cervical adenocarcinoma is necessary. Src is a proto-oncogene that is important in cancer progression. Dasatinib is a Src inhibitor that has been reported to be effective when used in combination with anticancer drugs. The present study aimed to confirm Src expression in human cervical adenocarcinoma cell lines and to determine the mechanism underlying the inhibitory effect of dasatinib on Src signaling in vitro. Western blot analysis was performed to investigate Src expression in cervical adenocarcinoma cell lines (HeLa and TCO-2 cells). The cells were cultured for 48 h with the addition of different concentrations of anticancer drugs (paclitaxel or oxaliplatin). Viable cell count was measured using a colorimetric (WST-1) assay. The concentrations of anticancer agents were fixed according to the results obtained, and the same experiments were performed using the drugs in combination with dasatinib at various concentrations to determine the concentrations that significantly affected the number of viable cells. The presence or absence of apoptosis was investigated using a caspase-3/7 assay. Signal transduction in each cell line was examined using western blotting. Src was activated in the two cell lines, and cell proliferation was significantly suppressed by each anticancer drug in combination with 10 µM dasatinib. Caspase-3/7 activity was also increased and Src signaling was suppressed by each anticancer drug in combination with dasatinib. In conclusion, Src is overexpressed in cervical adenocarcinoma cell lines, and dasatinib inhibits intracellular Src signaling and causes apoptosis. The results of the present study suggest that Src may be targeted in novel therapeutic strategies for cervical adenocarcinoma.

SFPQ, a multifunctional nuclear protein, regulates the transcription of PDE3A.

Phosphodiesterase 3A (PDE3A), a member of the cGMP-inhibited cyclic nucleotide phosphodiesterase (PDE) family, plays important roles in oocyte maturation and vascular smooth muscle cell proliferation. However, the molecular mechanisms that regulate PDE3A gene expression remain largely unknown. In the present study, we investigated the transcriptional regulation of PDE3A, and found that the splicing factor proline- and glutamine-rich (SFPQ) protein modulated PDE3A mRNA levels. Multiple transcription start sites (TSS1, 2, and 3) were identified within the first exon of PDE3A using 5′-rapid amplification of cDNA ends (RACE). Variable expression levels of three PDE3A variants were also observed in human tissues and HeLa cells. Several putative SFPQ-binding sites were identified upstream of the regulatory region of PDE3A-TSSs using ChIP sequencing (ChIP-seq). Serum-induced PDE3A expression was affected by increasing the amount of SFPQ binding to the upstream regulatory region of PDE3A. In addition, transcription of PDE3A was lower in human cervical adenocarcinoma cells compared with normal cervical tissue. Furthermore, overexpression of PDE3A induced sensitivity to anticancer therapeutic agent, 6-(4-(diethylamino)-3-nitrophenyl)-5-methyl-4,5-dihydropyridazin-3(2H)-one (DNMDP), in HeLa cells. Taken together, these results suggest that SFPQ functions as a transcriptional activator of PDE3A, which is involved in the regulation of DNMDP sensitivity, offering a novel molecular target for the development of anticancer therapies.

HPLC analysis, anti-oxidant activity of Genista ferox and its anti-proliferative effect in HeLa cell line

The prevention and treatment of the cancer using plants have attracted increasing interest. The present study was aimed to determine the phenolic compounds of Genista ferox using HPLC-TOF/MS and the anti-oxidant acti-vity associated with anti-cancer activity against human cervical adenocarcinoma (HeLa) cell line. Total anti-oxidant capacities of different extracts of G. ferox were assessed by DPPH assay, and their total phenolic and flavonoids contents measured by Folin–Ciocalteu and aluminum trichloride assays. The amounts of total phenolic (105.2 ± 0.6 – 308.5 ± 5.7 mg/g) of extract measured as gallic acid equivalent and flavonoids (8.1 ± 0.1 – 124.0 ± 0.7 mg/g) of extract measured as quercetin equivalent varied from chloroform to n-butanol extract of the two parts of the plant (leaf and stem). The ethyl acetate extract of G. ferox exhibited the most powerful effect on the DPPH scavenging activity with 94% from the leaf and 93% from the stem, while the chloroform extract from the leaf exhibited the most effective anti-proliferative activity against HeLa cell lines.
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Real-Time Cell Analysis of the Cytotoxicity of Origanum acutidens Essential Oil on HT-29 and HeLa Cell Lines.

The aim of this study was to investigate the cytotoxic and radical scavenging effects of the essential oil from Origanum acutidens (Hand-Mazz.) Ietswaart and to determine its chemical composition. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography/mass spectrometry (GC-MS). The cytotoxic effect of the essential oil on the growth of human colorectal adenocarcinoma (HT-29) and human cervical adenocarcinoma (HeLa) cell lines was investigated by xCELLigence Real Time Cell Analyzer instrument. Moreover, radical scavenging effect of the oil was determined by using DPPH method. The main constituents of the oil were found to be carvacrol (61.69%), p-cymene (17.32%), γ-terpinene (4.05%), and borneol (3.96%). The essential oil of O. acutidens exhibited significant cytotoxic effect against HeLa and HT-29 cell lines at the tested concentrations. The essential oil had moderate DPPH radical scavenging activity compared to butylated hydroxytoluene (BHT). The data in the present study clearly demonstrated inhibitory effect of the oil on two human cancer cell lines. According to these results and other reported studies, this observed high effect may be attributed to the presence of the carvacrol component in the oil.

Comparative studies on the antimicrobial and cytotoxic activities of Tanacetum vulgare L. essential oil and methanol extracts

Chemical composition of essential oil (EO) and methanol extracts (MEs) from different parts of Tanacetum vulgare L. plant was analyzed and investigated for potential biological activities and correlated with the main constituents detected in EO and MEs.The EO was characterized by a high content of oxygenated monoterpenes with trans-chrysanthenyl acetate as major compound. All MEs were characterized by neochlorogenic, 3,5-O-dicaffeoylquinic and caffeoylquinic acids. High phenolic content in MEs correlated to high antioxidant capacity, especially for roots. Tansy EO showed strong activity against the most of tested fungi and was efficient as bifonazole and ketoconazole, while the most sensitive bacteria were Gram-negative E. coli and E. cloacae. All MEs showed fungistatic and fungicidal effects against all of the eight tested fungi, but the antimicrobial activity was higher against Gram-positive bacteria. Additionally, shown for the first time, MEs of leaves and flowers exhibited a strong antiproliferative effect on human cervical adenocarcinoma (HeLa) cells, causing cell shrinkage and detachment.In the present study, the Tansy extracts and essential oil with low thujone content can provide a very promising and effective alternative in the field of antimicrobial applications and food preservation. The Tansy MEs possess a high antioxidant potential with phenolic acids being a major radical scavenging contributor with the proved antiproliferative activity to HeLa cells.

Influence of dynamic flow environment on nanoparticle-protein corona: From protein patterns to uptake in cancer cells

The fast growing use of nanoparticles (NPs) in biotechnology and biomedicine raises concerns about human health and the environment. When introduced in physiological milieus, NPs adsorb biomolecules (especially proteins) forming the so-called protein corona (PC). As it is the PC that mostly interacts with biological systems, it represents a major element of the NPs’ biological identity with impact on nanotoxicology, nanosafety and targeted delivery of nanomedicines. To date, NP-protein interactions have been largely investigated in vitro, but this condition is far from mimicking the dynamic nature of physiological environments. Here we investigate the effect of shear stress on PC by exposing lipid NPs with different surface chemistry (either unmodified and PEGylated) to circulating fetal bovine serum (FBS). PC formed upon in vitro incubation was used as a reference. We demonstrate that PC is significantly influenced by exposure to dynamic flow and that changes in PC composition are dependent on both exposure time and NP’s surface chemistry.Notably, alterations induced by dynamic flow affected cellular uptake of lipid NPs in both human cervical cancer (HeLa) and human breast adenocarcinoma (MCF7) cell lines.

Assessment of anti-cancerous potential of 6-gingerol (Tongling White Ginger) and its synergy with drugs on human cervical adenocarcinoma cells

The anti-cancerous activity of 6-gingerol extracted from Tongling White Ginger was investigated. 6-Gingerol inhibited the growth of HeLa cells with IC50 (96.32 μM) and IC80 (133.01 μM) and led to morphological changes, induced the cell cycle arrest in G0/G1-phase and ultimately resulted into apoptosis. Among cell cycle-related genes and proteins, the expression of cyclin (A, D1, E1) reduced, while of CDK-1, p21 and p27 showed slight decrease, except cyclin B1 and E1 (protein). Western blotting reported the induction of apoptosis with an increased Bax/Bcl-2 ratio, release of cytochrome c, cleavage of caspase-3, -8, -9 and PRPP in treated cells. 6-Gingerol activated AMPK, but inhibited PI3K/AKT phosphorylation with reduced P70S6K expression and also suppressed the mTOR phosphorylation. 6-Gingerol with 5-FU and Ptx resulted in 83.2% and 52% inhibition respectively, this synergy have stimulated apoptosis proteins more efficiently as compared to 6-Gingerol alone (10.75%) under in vitro conditions.

Lipophilic triphenylphosphonium derivatives enhance radiation-induced cell killing via inhibition of mitochondrial energy metabolism in tumor cells

It has recently been reported that radiation enhances mitochondrial energy metabolism in various tumor cell lines. To examine how this radiation-induced alteration in mitochondrial function influences tumor cell viability, various lipophilic triphenylphosphonium (TPP+) cation derivatives and related compounds such as 4-hydroxy-2,2,6,6-tetramethyl-1-oxy-piperidin (Tempol) with TPP+ (named “Mito-”) were designed to inhibit the mitochondrial electron transport chain. Mito-(CH2)10-Tempol (M10T) and its derivatives, Mito-(CH2)5-Tempol (M5T), Mito-(CH2)10-Tempol-Methyl (M10T-Me), Mito-C10H21 (M10), and C10H21-Tempol (10T), were prepared. In HeLa human cervical adenocarcinoma cells and A549 human lung carcinoma cells, the fractional uptake of the compound into mitochondria was highest among the TTP+ analogs conjugated with Tempol (M10T, M5T, and 10T). M10T, M10T-Me, and M10 exhibited strong cytotoxicity and enhanced X-irradiation-induced reproductive cell death, while 10T and M5T did not. Furthermore, M10T, M10T-Me, and M10 decreased basal mitochondrial membrane potential and intracellular ATP. M10T treatment inhibited X-ray-induced increases in ATP production. These results indicate that the TPP cation and a long hydrocarbon linker are essential for radiosensitization of tumor cells. The reduction in intracellular ATP by lipophilic TPP+ is partly responsible for the observed radiosensitization.

Glyco-functionalized dinuclear rhenium(i) complexes for cell imaging.

The design, synthesis and photophysical characterization of four new luminescent glycosylated luminophores based on dinuclear rhenium complexes, namely Glyco-Re, are described. The derivatives have the general formula [Re2(μ-Cl)2(CO)6(μ-pydz-R)] (R-pydz = functionalized 1,2-pyridazine), where a sugar residue (R) is covalently bound to the pyridazine ligand in the β position. Different synthetic pathways have been investigated including the so-called neo-glycorandomization procedure, affording stereoselectively glyco-conjugates containing glucose and maltose in a β anomeric configuration. A multivalent dinuclear rhenium glycodendron bearing three glucose units is also synthesized. All the Glyco-Re conjugates are comprehensively characterized and their photophysical properties and cellular internalization experiments on human cervical adenocarcinoma (HeLa) cells are reported. The results show that such Glyco-Re complexes display interesting bio-imaging properties, i.e. high cell permeability, organelle selectivity, low cytotoxicity and fast internalization. These findings make the presented Glyco-Re derivatives efficient phosphorescent probes suitable for cell imaging application.

Phytochemical analysis, antiproliferative and antioxidant activities of Chrozophora tinctoria: a natural dye plant

Context: Chrozophora tinctoria (L.) A. Juss. (Euphorbiaceae) is known as ‘dyer’s-croton’ and used to obtain dye substances. Recently, natural antioxidants and colorants have been of interest because of their safety and therapeutic effects.Objective: This study investigates the antiproliferative and antioxidant activities of the various extracts and fractions from C. tinctoria and analyzes their phytochemical contents.Materials and methods: The aerial parts of C. tinctoria were extracted with water, ethyl acetate, n-butanol, and methanol/chloroform. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The ethyl acetate extract (EA) was fractionated by flash chromatography. The extracts, fractions, and major phenolic compounds were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line at the concentrations of 5–100 μg/mL by using BrdU ELISA assay during 24 h of incubation. DPPH radical scavenging activities (5–150 μg/mL) and total phenolic contents of the samples were also evaluated.Results: 4-Hydroxybenzoic acid (268.20 mg/kg), apigenin-7-glucoside (133.34 mg/kg), and gallic acid (68.92 mg/kg) were the major components of EA. CT/E-F6 (IC50 = 64.59 ± 0.01 μg/mL) exhibited the highest antiproliferative activity. CT/E-F2 (IC50= 14.0 ± 0.0 μg/mL) and some fractions displayed higher radical scavenging activity compared to synthetic antioxidant BHT (IC50 = 23.1 ± 0.0 μg/mL). Among the main phenolics, gallic acid exhibited the highest antiproliferative and radical scavenging abilities (IC50 < 5 μg/mL).Conclusion: In this study, we have determined the biologically active fractions and their high effects may be attributed to the presence of gallic acid.

PLGA/SBA-15 mesoporous silica composite microparticles loaded with paclitaxel for local chemotherapy

In this work, high surface area mesoporous silica (SBA-15) was loaded with paclitaxel (taxol, PTX) and was further entrapped into poly(lactic acid-co-glycolic acid) (PLGA) microparticles (MPs). A modified solvent evaporation-emulsion method was used in order to formulate the composite microparticles with sizes of 8–12μm. PTX loaded SBA-15 as well as the PLGA/PTX-SBA-15 composites were characterized in terms of their morphology, crystal structure and thermal properties. Drug content, loading efficiency, particle size and the in-vitro drug release kinetics of the PLGA/PTΧ-SBA-15 microspheres were also investigated. The in vitro release studies were carried out using Simulated Body Fluid (SBF) at 37°C revealing that the prepared formulations present higher dissolution rate than pure PTX and sustained pattern which is ideal for anticancer carriers. Modeling and data analysis of the in vitro drug release was also investigated. It was also shown that all microparticles have low cytotoxicity in HUVE cells. Finally, it was found that drug loaded microparticles are very effective in Human Cervical Adenocarcinoma (HeLa) cells.

Methodology for monitoring gold nanoparticles and dissolved gold species in culture medium and cells used for nanotoxicity tests by liquid chromatography hyphenated to inductively coupled plasma-mass spectrometry

An analytical methodology based on coupling reversed-phase liquid chromatography (HPLC) to an inductively coupled plasma mass spectrometry (ICP-MS) has been developed for the characterization and identification of gold nanoparticles (AuNPs) and gold dissolved species (Au3+) in culture medium (Dulbecco's Modified Eagle Medium, DMEM) and HeLa cells (a human cervical adenocarcinoma cell line) used in nanotoxicity tests. The influence of the culture medium was also studied and the method applied for nanotoxicity tests. It was also observed that AuNPs can undergo an oxidation process in the supernatants and only a small amount of AuNPs and dissolved Au3+ was associated with cells. To evaluate the biological impact of AuNPs, a classical viability assay onto HeLa cells was performed using cellular media DMEM in the presence of increasing dosage of 10nm AuNPs. The results showed that 10nm AuNPs exhibit a slight toxic effect.

Eulophia macrobulbon – an orchid with significant anti-inflammatory and antioxidant effect and anticancerogenic potential exerted by its root extract

BackgroundThe Orchidaceae family is one of the largest families of flowering plants. Orchids are widely used for the traditional herbal medicine, acting as aphrodisiac, antisepic, antimicrobial, anti-cancer agent, etc. PurposeThis study was designed to elucidate the anti-inflammatory, antioxidant and cytotoxic potential of a 50% ethanolic extract of Eulophia macrobulbon roots (EME) in vitro, an orchid growing in Southern Asia. Furthermore, the main active compounds were isolated, and the bioactivity of the single constituents was determined. MethodsThe anti-inflammatory activity of EME and its compounds was evaluated by the secretion of pro- and anti-inflammatory cytokines and by the expression of inducible nitric oxide synthase (iNOS) in a lipopolysaccharide (LPS)-stimulated macrophage model, as determined by an enzyme linked immunosorbent assay (ELISA) and Western blot. Antioxidant activity was assessed using a DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) photometric assay. Cytotoxic effects were determined using a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-assay. ResultsEME and its compounds significantly reduced the production of the proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α), the expression of iNOS and subsequently increased the production of the anti-inflammatory cytokine interleukin 10 (IL-10) in LPS-stimulated macrophages. Additionally it could be demonstrated that EME is rich in radical scavengers. Furthermore, EME and its components showed notable cytotoxic effects on the human cervical adenocarcinoma cell line HeLa, the human colorectal adenocarcinoma cell line CaCo-2 and the human breast adenocarcinoma cell line MCF-7. The most active constituents were identified as 4-methoxy-9,10-dihydro-2,7-phenanthrenediol (8), 4-methoxy-2,7-phenanthrenediol (9), 1,5-dimethoxy-2,7-phenanthrenediol (10), 1,5,7-trimethoxy-2,6-phenanthrenediol (11), 1-(4-hydroxybenzyl)-4,8-dimethoxy-2,7-phenanthrenediol (15). ConclusionBased on this data, EME provides various beneficial anti-inflammatory, antioxidant and cytotoxic attributes and may be used as herbal remedy in the pharmaceutical or food industries.