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Myxoma virus (MyxV) is a rabbit-specific poxvirus. However, its ability to selectively target tumor cells has established it as a safe and effective anticancer therapy. To strengthen its preclinical efficacy, transgenes that can prolong cancer cell infection and enhance anti-tumor effector functions are currently being investigated. We engineered MyxV armed with CD47, to turn on a 'do not eat me' signal within infected cells with actively replicating viruses, and with IFN-γ to further activate host immune anticancer responses. Tumor suppressive activities were significantly enhanced by the dual-armed MyxV_CD47/IFN-γ compared to parental MyxV or single-armed MyxV_CD47 or MyxV_IFN-γ. In addition, significant increases in IFN-γ+ CD8+T-cells and CD4+ T-cells populations within tumor-infiltrating lymphocytes (TIL) were observed after MyxV_CD47/IFN-γ treatment. Notably, all groups treated with MyxV showed a marked reduction in Foxp3+ CD4+ regulatory T-cells (Tregs) within TIL. We also show that MyxV infection induces PD-L1 up-regulation in cancer cells, and combinational treatment of MyxV with anti-mouse PD-L1 antibodies (αPD-L1) further controlled tumor burden and increased survival in the syngeneic melanoma model B16F10. Our data demonstrate that a CD47 and IFNγ dual-armed MyxV is an effective oncolytic viral immunotherapeutic. These findings strongly support further preclinical investigations to develop next-generation MyxV-based immunotherapy approaches.
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Metabolic variations, antioxidant potential and cytotoxic effects were investigated in the different plant parts like the leaf, stem, flower, pod, and root of C. majus L. using spectroscopic and chromatographic methods. Total phenolics and flavonoids were studied in the different parts of C. majus L., leaf showed higher flavonoid content (137.43 mg/g), while the pod showed the highest phenolic (23.67 mg/g) content, when compared with the stem, flower and root. In the ABTS antioxidant assay, the flower extract showed 57.94% effect, while the leaf, pod and root extract exhibited 39.10%, 36.08% and 28.88% activity, respectively. The pod and leaf extracts demonstrated the potential effect, exhibiting 45.46 and 41.61% activity, respectively, for the DPPH assay. Similar to the phosphomolybdenum assay, the flower revealed higher antioxidant activity (46.82%) than the other plant parts. The in vitro SRB assay facilitated evaluation of the cytotoxic effect against the HeLa and CaSki human cervical cancerous cells. The extract displayed dose-dependent inhibitory effect on both the cell lines. The highest cytotoxic effect was observed in the pod and flower extracts post 48 h of exposure at 1000 µg/mL. The results of C. majus L. offered new insights in the preliminary steps regarding the development of a high value product for phytomedicine applications though promising metabolic variations with antioxidant and anticancer potentials.
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Antineoplásicos/farmacología , Antioxidantes/farmacología , Chelidonium/metabolismo , Flavonoides/análisis , Células HeLa , Humanos , Polifenoles/análisisRESUMEN
The extract of Phyllodium (P.) elegans was investigated for its anti-cancer properties on brain astroglioma cells (U251-MG), colorectal carcinoma cells (HCT116), and malignant melanoma cells (A375). P. elegans methanolic extract (PeME) showed cytotoxicity on all three cancer cell lines tested. The cell viability assay revealed that PeME significantly reduced the viability of these cells. Clear apoptotic features such as cellular morphology, cell shrinkage, and augmentation of dead cells were observed. Flow cytometry and fluorescence staining techniques confirmed the apoptotic property of PeME. In vitro scratch invasion assay showed that cell migration rate was significantly reduced. Fluorescence microscopic studies using 4',6-diamidino-2-phenylindole staining showed early and late signs of apoptosis after PeME treatment. Upon PeME stimulation, activation of caspase-3/-9 and Mu-2-related death-inducing gene (MUDENG, MuD) was observed by western blot analysis. JC-1 staining analysis by flow cytometry showed that PeME depolarized the mitochondria membrane potential (MMP). Collectively, these findings, for the first time, point to the fact that PeME has anti-cancer properties against brain, colon, and skin cancer cell lines by depolarizing the MMP and activating apoptotic signaling through the activation of caspase-3/-9 as well as MuD. This is the first report reporting the anticancer activity of this specific plant extract.[Figure: see text].
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Fabaceae/química , Extractos Vegetales/farmacología , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Caspasas/genética , Caspasas/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Humanos , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
Dianthus superbus (DS) is a traditional medicinal herb well known for its medicinal and therapeutic potential and widely distributed in various Asian countries. The ethyl acetate (EA), butanol (Bu) and distilled water (DW) extracts of DS assessed for extraction of bioactive compounds and their biological activities. The chemical analysis was done using LC-MS/MS and antioxidant, anticancer and antiviral activities were determined. EA extracts showed strong anticancer activity with IC50 of 9.5, 13.8 and 69.9⯵g/mL on SKOV, NCL-H1299 and Caski cancer cell lines, respectively. The Bu extracts exhibited strongest antiviral activity with respect to both influenza A and B viruses with IC50 values of 4.97 and 3.9⯵g/mL, respectively. Also the metabolic profile for EA, Bu and DW extracts shows high variations and influence precisely the antioxidant, anticancer and antiviral properties. The quercetin 3- rutinoside and isorhamnetin 3- glucoside showed higher neuraminidase inhibition activity in dose dependent manner. Molecular docking study revealed that flavonol glycosides have higher binding activities towards influenza polymerase membrane glycoprotein. Correlation study showed that flavonol glycosides were linked to anti-influenza activity and cyclic peptides with anticancer activities. This study provides vital information for effective utilization of DS for medicinal, food and therapeutic purposes.
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Antineoplásicos/farmacología , Antioxidantes/farmacología , Antivirales/farmacología , Dianthus/química , Flavonoles/farmacología , Glicósidos/farmacología , Animales , Antineoplásicos/aislamiento & purificación , Antioxidantes/aislamiento & purificación , Antivirales/aislamiento & purificación , Línea Celular Tumoral , Perros , Flavonoles/aislamiento & purificación , Glicósidos/aislamiento & purificación , Humanos , Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza B/efectos de los fármacos , Células de Riñón Canino Madin Darby , Simulación del Acoplamiento Molecular , Neuraminidasa/antagonistas & inhibidores , Proteínas Virales/antagonistas & inhibidoresRESUMEN
In this study, we investigated the antiviral activity of newly synthesized berberine derivatives (BD) against influenza virus infection using several strains in in vitro and in silico. The CPE reduction, pre-incubation, NA activity inhibition and molecular docking assays were used for antiviral evaluation. The anti-influenza activities of BDs were stronger than plant-derived pure commercial berberine, and some of the BDs were more potent than control drug Oseltamivir. The cytotoxicity level was observed in the range 63.16-1639µg/mL for synthesized BDs. Additionally, BDs were detected as able to block influenza viral particles. We targeted neuraminidase one of the influenza surface protein for further probing. Moreover, BDs registered competitive NA inhibition activity comparing with Oseltamivir. The active site of viral NA subunit was fully blocked by BD as the same location as Oseltamivir. The binding energies between influenza NA subunit and BD-5 were higher than Oseltamivir. More H-bonds and NA residues were occupied by BD for stronger binding ability than Oseltamivir. These results indicated that BD inhibits various strains of influenza virus by blocking of viral NA subunit.
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Antivirales/farmacología , Berberina/farmacología , Descubrimiento de Drogas , Inhibidores Enzimáticos/farmacología , Neuraminidasa/antagonistas & inhibidores , Orthomyxoviridae/efectos de los fármacos , Antivirales/síntesis química , Antivirales/química , Berberina/síntesis química , Berberina/química , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Humanos , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Neuraminidasa/metabolismo , Orthomyxoviridae/enzimología , Relación Estructura-ActividadRESUMEN
Aspartame is widely used artificial sweeteners as food additives. Several researchers have pointed that the controversial report on the use of aspartame over more than decades. Omega-3 fatty acids are essential and unsaturated fatty acids, and it plays a remarkable role in vision, intelligence, neural development, and metabolism of neurotransmitters. Therefore, the present study was aimed to investigate the effect of omega-3 fatty acids on aspartame treated renal cells. Experimental groups were divided into three such as sham control, aspartame treated, and aspartame with omega-3 fatty acids. Cell viability was determined by sulforhodamine-b assay and flow cytometric analysis. The experimental results showed that the aspartame induced altered cell viability were reduced following treatment of aspartame with omega-3 fatty acids. Altered cell morphology was recovered by omega-3 fatty acids. DNA damage appeared in the highest concentration of aspartame used in this study. DNA damage characteristics such as comet tail and tiny head sections did not appear in the omega-3 fatty acids treated cells. Several microvilli and vesicular structures were found in aspartame treated cells. Altered morphology such as rounding, microvilli, and formation of dome-like structures did not appear in the omega-3 fatty acids with aspartame treated cells. Caspase-3 mRNA and protein expression were increased in aspartame treated cells, and these levels were reduced following omega-3 fatty acids treatment. Taking all these data together, it is suggested that the omega-3 fatty acids may be a therapeutic agent to reduce the aspartame induced biochemical and morphological alterations in normal renal cells. © 2017 BioFactors, 43(6):847-857, 2017.
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Aspartame/toxicidad , Ácidos Grasos Omega-3/farmacología , Sustancias Protectoras/farmacología , Animales , Caspasa 3/genética , Caspasa 3/metabolismo , Supervivencia Celular/efectos de los fármacos , Ensayo Cometa , Fragmentación del ADN/efectos de los fármacos , Perros , Expresión Génica , Células de Riñón Canino Madin Darby , ARN Mensajero/genética , ARN Mensajero/metabolismo , Rodaminas/químicaRESUMEN
The present study investigates the anticancer activity of Thidiazuron (TDZ). Anticancer activity of TDZ was evaluated in cervical carcinoma cells (HeLa cells). Sulforhodamine-B (SRB) assay indicates that TDZ was about 100 times more toxic to the cancer cell than normal cells. TUNEL assay showed TDZ induced DNA damage in tumor cells. The loss of mitochondrial membrane potential (MMP) in cancer cells was observed following TDZ treatment. The Bax and bcl-2 gene expression ratio are highly responsible for the regulation of MMP balance, and these ratio was significantly altered following TDZ treatment. The p53 and caspase-3 expressions were increased in cancer cells following treatment. Caspase-3 activation is the key factor for apoptosis. Cytotoxicity of TDZ on HeLa cells was 100 times higher than normal kidney cell (MDCK cells). Moreover, the anticancer activity of TDZ was tested by DNA damage, mitochondrial dysfunction, some gene expression and caspase-3 inhibition in silico. TDZ detected has higher ability on early apoptosis of cancer cell through DNA damage. Additionally, cancer cellular MMP was significantly reduced under inoculation of TDZ. In silico assay confirmed that TDZ was able to bind with the active site of the capase-3 protein. Therefore, taking all these data together it is suggested that the TDZ may be a potential agent to act against cervical cancer cells.
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Daño del ADN/efectos de los fármacos , Compuestos de Fenilurea/toxicidad , Tiadiazoles/toxicidad , Animales , Sitios de Unión , Caspasa 3/química , Caspasa 3/genética , Caspasa 3/metabolismo , Perros , Femenino , Expresión Génica/efectos de los fármacos , Células HeLa , Humanos , Células de Riñón Canino Madin Darby , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microscopía Fluorescente , Simulación del Acoplamiento Molecular , Compuestos de Fenilurea/química , Compuestos de Fenilurea/metabolismo , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/metabolismo , Termodinámica , Tiadiazoles/química , Tiadiazoles/metabolismo , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patología , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
ß-Alanine is a non-essential amino acid and presents as a major component of various sports supplements. It is a non-proteogenic amino acid, formed in vivo by degradation of carnosine, anserine, balenine, and dihydrouracil. The present study was aimed at investigating the anti-tumor effects of ß-alanine in renal and cervical tumor cells. Sulforhodamine-B assay and flow cytometric analysis were used to measure cell viability. Lactate dehydrogenase (LDH) expression was analyzed using FITC-conjugated fluorescent antibody. The cellular adenosine triphosphate (ATP) content was measured using bioluminescence method. Cell migration was determined by the simple standard-scratch method. ß-Alanine reduced renal and cervical cell growth significantly. Percentage of inhibition of renal and cervical tumor cells was increased at higher concentration of ß-alanine. LDH expression and ATP content were significantly reduced in renal and cervical tumor cells in a dose-dependent manner. Renal and cervical tumor cell migration were significantly reduced following 10 and 100 mM of ß-alanine treatment. In our study, ß-alanine exerts no significant effect on normal MDCK cells except a marginal effect at the highest concentration (100 mM). In summary, our experimental data suggest that ß-alanine may be a potential anti-tumor agent exhibiting several anti-cancer effects in renal and cervical tumor cells.
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Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Neoplasias Renales/metabolismo , Neoplasias del Cuello Uterino/metabolismo , beta-Alanina/farmacología , Femenino , Humanos , Neoplasias Renales/tratamiento farmacológico , Neoplasias Renales/patología , L-Lactato Deshidrogenasa/metabolismo , Células Tumorales Cultivadas , Neoplasias del Cuello Uterino/tratamiento farmacológico , Neoplasias del Cuello Uterino/patologíaRESUMEN
Influenza virus has had a high rate of antigenic shift and drift that causes significant morbidity and mortality in humans and animals. The lack of excellent pharmacological treatment underlines the importance of the development of the novel antiviral drugs. We investigated the anti-influenza A and B viruses of 2,4-dichlorophenoxyacetic acid (2,4-D), which is the synthetic analog to auxin and is used as a popular herbicide in the agricultural practices. 2,4-D was evaluated using a cytopathic effect reduction method; assay results showed that 2,4-D possessed strong anti-influenza A and B viruses inhibiting the formation of a visible cytopathic effect. Influenza viral RNA expression was performed by quantitative real-time polymerase chain reaction. 2,4-D also inhibited virus replication in the early stage of influenza virus infection without direct interaction with virus particles. Additionally, 2,4-D significantly inhibited various factors occur during influenza virus infection as the acidic vesicular formation and reactive oxygen species production. Moreover, 2,4-D represented no cytotoxicity in normal kidney cell. Therefore, these findings provide an understanding of the mechanism and efficient use of 2,4-D in pharmacological applications against influenza virus infection.
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Ácido 2,4-Diclorofenoxiacético/farmacología , Antivirales/farmacología , Autofagia/efectos de los fármacos , Regulación Viral de la Expresión Génica , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Replicación Viral/efectos de los fármacos , Animales , Perros , Interacciones Huésped-Patógeno , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/metabolismo , Subtipo H3N2 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/metabolismo , Células de Riñón Canino Madin Darby , Neuraminidasa/antagonistas & inhibidores , Neuraminidasa/genética , Neuraminidasa/metabolismo , Oseltamivir/farmacología , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Viral/antagonistas & inhibidores , ARN Viral/genética , ARN Viral/metabolismo , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/metabolismo , Proteínas Virales/antagonistas & inhibidores , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
Seabuckthorn is a medicinal plant that is used to prevent cold. It was tested for its metabolic content followed by activity against cancer and virus. The metabolic distribution of different polarity solvent extractions from the leaves was analyzed by LC-MS/MS. Flavonol glycoside contents in EA and Bu extracts were higher than MeOH and DW was observed. MeOH and EA extracts recorded high activity against influenza A/PR virus with IC50 of 7.2 µg/mL and 10.3 µg/mL compared with known drug Oseltamivir of 60.3 µg/mL. A similar trend showed in influenza A/Victoria virus. In case of influenza B viruses such as B/Lee and B/Maryland, EA extract (2.87 µg/mL and 4.5 µg/mL of IC50) emerged strongest among other extracts and Oseltamivir (103.73 µg/mL and 71.6 µg/mL). Each extract showed potent anticancer activities. Interestingly, Bu extract showed stronger anticancer activity against human cancer cells such as NCL-H1299, HeLa, SKOV and Caski (8.2 µg/mL, 8.6 µg/mL, 18.2 µg/mL and 9.2 µg/mL of IC50) respectively. Correlation study reveals that aglycones and flavonol mono-glycosides highly correlated with anti-influenza activities but not correlated with anticancer activities. Reversely, di-glycosides and tri-glycosides have a high correlation with cytotoxic effect with both normal and cancer cells. Therefore, this study provides significant information concerning Seabuckthorn for further medicinal drug development.
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Mushrooms that have been restricted to fresh markets have now entered commercial and pharmaceutical markets. However, not much research has been targeted on testing the efficiency of these commercialized mushroom powders or capsules. For the first time, efforts were made to study the bioactive properties and antimicrobial properties of four predominant mushroom capsules available for commercial purposes. Then, these commercial mushroom powders were downsized to ultrasized fine powders by sonication and then their properties were compared against the commercialized ones. The results indicated that the bioactive properties and the antioxidant properties of these powders when used as marketed capsules was very less. It was following ultrasonication assisted size reduction that the cumulative bioactivity related properties got accelerated. Micro size reduction of the mushroom powders lead to significant enhancement of antiviral properties compared to antibacterial and antifungal properties. This work demonstrates that commercialization of mushroom as powders could realize higher impact through sonication assisted ultrasizing and even nanosizing.
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Antiinfecciosos/farmacología , Ganoderma/química , Pleurotus/química , Reishi/química , Tricholoma/química , Animales , Aspergillus niger/efectos de los fármacos , Aspergillus niger/crecimiento & desarrollo , Perros , Tecnología de Alimentos/métodos , Humanos , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H1N1 del Virus de la Influenza A/crecimiento & desarrollo , Células de Riñón Canino Madin Darby , Pruebas de Sensibilidad Microbiana , Tamaño de la Partícula , Penicillium/efectos de los fármacos , Penicillium/crecimiento & desarrollo , Polvos , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Sonicación , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/crecimiento & desarrolloRESUMEN
Aspartame is an artificial sweetener used as an alternate for sugar in several foods and beverages. The study reports that consumption of aspartame containing product could lead to cancer. However, the effect of aspartame on apoptosis process in cancer is not yet understood clearly. HeLa cells were exposed to different concentrations (0.01-0.05 mg/ml) of aspartame for 48 h. Cytotoxicity of aspartame on cancer cells was determined by SRB assay. The result indicates no significant changes on cell viability. Aspartame suppresses apoptosis process in cancer cells by down-regulation of mRNA expression of tumor suppressor gene p53, and pro-apoptotic gene bax. It up-regulates anti-apoptotic gene bcl-2 mRNA expression. In addition, Ki 67 and PCNA mRNA, and protein expressions were determined. Taking all these together, we conclude that aspartame may be a potent substance to slow-down the apoptosis process in HeLa cells. Further works are ongoing to understand the biochemical and molecular mechanism of aspartame in cancer cells.
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Natural substances have been attracted several researchers in the recent years, because of its potential antioxidant, anti-inflammatory and anti-cancer properties. We have investigated the effect of carnosine on cell viability, apoptosis, DNA damage, reactive oxygen species (ROS) and caspase 3 enzyme expression in human cervical carcinoma and Madin-Darby Kidney Cells (MDCK) cells. Carnosine inhibited cancer cell growth up to 23%. ROS level was increased up to 30 and 31% in MDCK and HeLa cells respectively. Tunnel assay showed 42 and 14% of positive apoptotic cells in cancer and normal cells respectively. The alteration in mitochondrial and nuclear morphology was determined. The extended lace-like network of normal mitochondria found in control cells. Carnosine treatment significantly altered the mitochondrial morphology of normal cervical carcinoma cell. Mitochondria were condensed clump structures in carnosine treated cancer cells. Carnosine reduced the number of colonies of cervical carcinoma cells. Caspase 3 expression was corresponded to the appearance of immunofluorescence in the cytoplasm. Caspase 3 expression was gradually increased in cervical carcinoma cells. In Silico, docking study was performed to recognize the binding activity of carnosine against a subunit of the caspase 3, and carnosine was able to bind to the drug binding pocket of caspase 3. The glide energy is -5.2 kcal/mol, suggesting the high binding affinity of carnosine to caspase 3. Taking all these data together, the natural dipeptide L-carnosine could be a suitable antiproliferative agent in cervical carcinoma cells. Copyright © 2016 John Wiley & Sons, Ltd.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Carcinoma/tratamiento farmacológico , Carnosina/farmacología , Neoplasias del Cuello Uterino/tratamiento farmacológico , Animales , Caspasa 3/biosíntesis , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Perros , Femenino , Técnica del Anticuerpo Fluorescente , Células HeLa , Humanos , Células de Riñón Canino Madin Darby , Mitocondrias/fisiología , Simulación del Acoplamiento Molecular , Especies Reactivas de Oxígeno/metabolismoRESUMEN
A metal oxide nanoparticle has been widely investigated for its potential use in the biomedical application. The present study investigates the cytotoxicity of ZnO nanoparticle in human cervical carcinoma cells. Cell viability was determined, and it showed the possible cytotoxic effect of ZnO nanoparticles. The characteristic apoptotic features such as rounding and loss of adherence were observed in the treated cells. Fluorescence and Confocal Laser Scanning Microscope (CLSM) studies have showed reduced nuclear volume and condensed cytoplasm. The mRNA expression of apoptotic gene p53 and caspase 3 was up-regulated following ZnO nanoparticle exposure, which confirms the occurrence of apoptosis at the transcriptional level. Reactive oxygen species (ROS) was increased in a dose-dependent manner, and initiate lipid peroxidation of the liposomal membrane, which in turn regulate several signaling pathways and influencing the cytokinetic movements of cells. ZnO nanoparticles showed a dynamic cytotoxic effect in cervical carcinoma cells. ZnO nanoparticle might induce the apoptosis through increased intracellular ROS level. Moreover, up-regulated apoptotic gene expression confirms the occurrence of apoptosis. Taking all these data together, it may be concluded that ZnO nanoparticle may exert cytotoxicity on HeLa cell through the apoptotic pathway, implies the probable utility of ZnO nanoparticle in the cancer treatment and therapy.
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Antineoplásicos/farmacología , Nanopartículas del Metal , Neoplasias del Cuello Uterino/patología , Óxido de Zinc/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Femenino , Expresión Génica , Células HeLa , Humanos , Especies Reactivas de Oxígeno/metabolismo , Espectrofotometría UltravioletaRESUMEN
Aspartame is used as an artificial sweetener in more than 6000 food varieties. The present study aims to determine the effects of aspartame at various concentrations on the cell viability, morphology, ROS level and DNA of human cervical carcinoma cells over two time periods of exposure. The effects of aspartame on HeLa cell viability were investigated using the sulphorhodamine-B assay (SRB assay) and flow cytometry. Alkaline comet assay was carried out to determine the possible DNA damage induced by aspartame. Mitochondria-derived reactive oxygen species (ROS) were determined using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Fluorescence microscopy was used to determine the presence of apoptotic and necrotic cells following aspartame treatment. Cell viability was significantly altered following a higher concentration of aspartame exposure. Mitochondria-derived ROS increased at higher concentrations of aspartame exposure. Exposure to 10 mM and 20 mM of aspartame induced DNA fragmentation. Apoptotic and necrotic bodies were found in the range of 1-20 mM aspartame exposure. Exposure to high concentrations of aspartame may alter cell viability and morphology, and it may induce ROS generation and DNA damage in cervical carcinoma cells.
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Silver nanoparticles (AgNPs) have well-known anti-bacterial properties and have been widely used in daily life as various medical and general products. There is limited information available on the cytotoxicity of AgNPs. Therefore, the present study aimed to investigate the cytotoxicity of AgNPs in HeLa cells. Cytotoxicity and apoptosis have been observed in the AgNPs treated in the HeLa cells. Sulphorhodamine-B assay (SRB assay) showed the cytotoxic effect in the AgNP-treated HeLa cells. Inverted microscope, fluorescence microscope, and confocal laser scanning microscope (CLSM) analyses showed the apoptosis-induced morphological changes such as rounding in shape, nuclear fragmentation, cytoplasm reduction, loss of adhesion, and reduced cell volume. Necrosis and apoptosis were observed in the AgNP-treated HeLa cells by DNA fragmentation study. Mitochondria-derived reactive oxygen species (ROS) have increased in AgNP-treated HeLa cells. Up-regulation of messenger RNA (mRNA) expression of p53, bax, and caspase 3 were found in AgNP-treated HeLa cells. Caspase 3 enzyme activity was found to increase in AgNP-treated HeLa cells. The AgNPs showed the right cytotoxic effect in cervical carcinoma cells. Our results suggest that metal-based nanoparticles might be a potential candidate for the treatment of cervical cancer.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , Nanopartículas del Metal , Plata/farmacología , Neoplasias del Cuello Uterino/tratamiento farmacológico , Citotoxinas/farmacología , Relación Dosis-Respuesta a Droga , Femenino , Células HeLa , Humanos , Especies Reactivas de Oxígeno/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patologíaRESUMEN
Hormonal regulation of cell growth and development, tissue morphology, metabolism and physiological function in animals and man is a well-established knowledge domain in modern biological science. The present study was carried out to investigate the structural stability of hexokinase when exposed to diabetic levels of glucose and its binding efficiency. The fluorescence study indicated that 28-homobrassinolide was able to protect or restore the native structure of hexokinase. Proteins are synthesized and fold into the native form to become active. The inability of a protein molecule to remain in its native form is called as protein misfolding and this is because of several factors. Protein aggregation and misfolding are known to play a critical role in several human diseases including diabetes. Homobrassinolide interaction with hexokinase was studied by UV-Vis spectrophotometer and fluorescence spectrophotometer. Results were suggested that the denatured hexokinase was renatured upon binding with homobrassinolide. In silico, docking study was performed to recognize the binding activity of homobrassinolide against a subunit of the glucokinase, and homobrassinolide was able to bind to the drug binding pocket of glucokinase. The glide energy is -7.1 kcal/mol, suggesting the high binding affinity of homobrassinolide to glucokinase. Overall, these studies predict that the phytohormone 28-homobrassinolide would function as an anti-diabetic when present in human and animal diet by augmenting the hexokinase enzyme activity in the animal cell. Copyright © 2015 John Wiley & Sons, Ltd.
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Colestanonas/farmacología , Hexoquinasa/química , Hexoquinasa/efectos de los fármacos , Hipoglucemiantes/farmacología , Animales , Sitios de Unión , Estabilidad de Enzimas/efectos de los fármacos , Humanos , Modelos Moleculares , Simulación del Acoplamiento Molecular , Unión Proteica , Desnaturalización Proteica/efectos de los fármacos , Ingeniería de ProteínasRESUMEN
Cellular and physiological responses to the degradation products of titanium implants are key indicators to determine the quality of biocompatibility of implant devices. The present study investigated titanium dioxide (TiO2) nanoparticle-induced cytotoxicity, apoptotic morphological modification, and apoptotic-related gene expressions in the human cervical carcinoma cells. TiO2 nanoparticle-induced cytotoxicity on cancer cells was determined by the sulphorhodamine-B assay. Apoptotic morphological modification such as nuclear fragmentation, rounding, cytoplasm shrinkage, loss of adhesion, and reduced cell volume were observed by an inverted, fluorescence, and confocal laser scanning microscope (CLSM). The DNA fragmentation study showed the occurrence of necrosis and apoptosis in nanoparticle-treated cells. The qPCR study showed the increased p53 and bax mRNA expression in the nanoparticle-treated cells compared to control. In addition, caspase 3 activity was increased in nanoparticle-treated cells, which indicates the increased auto-catalysis. Taking all these data together, it may suggest that TiO2 nanoparticle could inhibit the growth of HeLa cells.
Asunto(s)
Nanopartículas/química , Nanopartículas/uso terapéutico , Titanio/farmacología , Titanio/uso terapéutico , Neoplasias del Cuello Uterino/tratamiento farmacológico , Neoplasias del Cuello Uterino/patología , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Células HeLa , Humanos , Relación Estructura-Actividad , Titanio/química , Células Tumorales CultivadasRESUMEN
Variations in antioxidant and anti-viral activities (against Influenza AP/R/8 (H1N1) virus) between the leaves and stem bark of selected medicinal plants were studied. Malin Darby canine kidney (MDCK) cells were used for the viral infection and the antiviral activity of the extracts was studied using sulphorhodamine B (SRB) assay. The stem bark of the plants including Strychnos minor, Diotacanthus albiflorus, Strychnos nux-vomica and Chloroxylon swietenia showed higher flavonoid contents as well as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activity when compared with their leaves. In case of 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity, the stem bark of S. nux-vomica and leaf extract of C. swietenia showed the highest activity. Based on the IC50 values, the stem bark extracts of Cayratia pedata (20.5 µg/mL) and S. minor (22.4 µg/mL) showed high antiviral activity. In the mean-time S. nux-vomica, C. swietenia and C. swietenia bark extracts showed cytotoxicity to the MDCK cells. When comparing the stem bark and leaves the content of gallic acid, ferulic acid, o-coumaric acid, total flavonoids (TFC) and total phenols (TPC) was higher in stem bark and hence their anti-viral activity was high. Further study based on the metabolites against H1N1 can reveal the potential of therapeutic compounds against the viral disease.
RESUMEN
Antiviral activity against H1N1 influenza was studied using ethnic medicinal plants of South India. Results revealed that Wrightia tinctoria (2.25 µg/ml) was one of the best antidotes against H1N1 virus in terms of inhibitory concentration of 50% (IC50) whereas the control drug Oseltamivir showed 6.44 µg/ml. Strychnos minor, Diotacanthus albiflorus and Cayratia pedata showed low cytotoxicity (>100) to the MDCK (Malin darby canine kidney) cells by cytotoxicity concentration of 50% (CC50) and possessed antiviral activity suggesting that these plants can be used as herbal capsules for H1N1 virus. W. tinctoria and S. minor showed high therapeutic indexes (TI) such as 12.67 and 21.97 suggesting that those plants can be used for anti-viral drug development. The CC50 values of Eugenia singampattiana (0.3 µg/ml), Vitex altissima (42 µg/ml), Salacia oblonga (7.32 µg/ml) and Salacia reticulata (7.36 µg/ml) resulted in cytotoxicity of the MDCK cells, due to their high phenolic content. Findings from this study state that the plant W. tinctoria can be a potent source for third generation anti-viral drug development against H1N1.