RESUMEN
Lectins are proteins that reversibly bind to carbohydrates and are commonly found across many species. The Banana Lectin (BanLec) is a member of the Jacalin-related Lectins, heavily studied for its immunomodulatory, antiproliferative, and antiviral activity. In this study, a novel sequence was generated in silico considering the native BanLec amino acid sequence and 9 other lectins belonging to JRL. Based on multiple alignment of these proteins, 11 amino acids of the BanLec sequence were modified because of their potential for interference in active binding site properties resulting in a new lectin named recombinant BanLec-type Lectin (rBTL). rBTL was expressed in E. coli and was able to keep biological activity in hemagglutination assay (rat erythrocytes), maintaining similar structure with the native lectin. Antiproliferative activity was demonstrated on human melanoma lineage (A375), evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). rBTL was able to inhibit cellular growth in a concentration-dependent manner, in an 8-h incubation, 12 µg/mL of rBTL led to a 28.94% of cell survival compared to cell control with 100%. Through a nonlinear fit out log-concentration versus biological response, an IC50% of 3.649 µg/mL of rBTL was determined. In conclusion, it is possible to state that the changes made to the rBTL sequence maintained the structure of the carbohydrate-binding site without changing specificity. The new lectin is biologically active, with an improved carbohydrate recognition spectrum compared to nBanLec, and can also be considered cytotoxic for A375 cells.
Asunto(s)
Escherichia coli , Lectinas , Humanos , Animales , Ratas , Lectinas/genética , Lectinas/farmacología , Escherichia coli/genética , Lectinas de Plantas/genética , Lectinas de Plantas/farmacología , Lectinas de Plantas/química , Secuencia de Aminoácidos , CarbohidratosRESUMEN
Preservative treatments increase the durability of wood, and one of the alternative treatments involves the use of chromated copper arsenate (CCA). Due to the toxicity of CCA, the disposal of CCA-treated wood residues is problematic, and burning is considered to provide a solution. The ecotoxicological potential of ash can be high when these components are toxic and mutagenic. The aim of this study was to evaluate the toxicity and genotoxicity of bottom ash leachates originating from CCA-treated wood burning. Physical-chemical analysis of the leachates revealed that in treated wood ashes leachate (CCA-TWBAL), the contents of arsenic and chromium were 59.45 mg.L-1 and 54.28 mg.L-1, respectively. In untreated wood ashes leachate (UWBAL), these contents were 0.70 mg.L-1 and 0.30 mg.L-1, respectively. CCA-TWBAL caused significant toxicity in Lactuca sativa, Allium cepa, and microcrustacean Artemia spp. (LC50 = 12.12 mg.mL-1). Comet assay analyses using NIH3T3 cells revealed that concentrations ranging from 1.0 and 2.5 mg.mL-1 increase the damage frequency (DF) and damage index (DI). According to MTT assay results, CCA-TWBAL at concentrations as low as 1 mg.mL-1 caused a significant decrease in cellular viability. Hemolysis assay analyses suggest that the arsenic and chromium leachate contents are important for the ecotoxic, cytotoxic, and genotoxic effects of CCA-TWBAL.
Asunto(s)
Antineoplásicos , Arsénico , Eliminación de Residuos , Animales , Arseniatos/química , Arseniatos/toxicidad , Arsénico/análisis , Cromo/análisis , Cobre/química , Daño del ADN , Ratones , Células 3T3 NIH , Eliminación de Residuos/métodos , Madera/químicaRESUMEN
The sodium valproate has been largely used as an anti-epilepsy drug and, recently, as a putative drug in cancer therapy. However, the treatment with sodium valproate has some adverse effects. In this sense, more effective and secure complexes than sodium valproate should be explored in searching for new active drugs. This study aims to evaluate the cytotoxicity of sodium valproate, mixed ternary mononuclear Cu(II) complexes based on valproic acid (VA) with 1,10-phenanthroline (Phen) or 2,2'- bipyridine (Bipy) ligands - [Cu2(Valp)4], [Cu(Valp)2Phen] and [Cu(Valp)2Bipy] - in yeast Saccharomyces cerevisiae, proficient or deficient in different repair pathways, such as base excision repair (BER), nucleotide excision repair (NER), translesion synthesis (TLS), DNA postreplication repair (PRR), homologous recombination (HR) and non-homologous end-joining (NHEJ). The results indicated that the Cu(II) complexes have higher cytotoxicity than sodium valproate in the following order: [Cu(Valp)2Phen] > [Cu(Valp)2Bipy] > [Cu2(Valp)4] > sodium valproate. The treatment with Cu(II) complexes and sodium valproate induced mutations in S. cerevisiae. The data indicated that yeast strains deï¬cient in BER (Ogg1p), NER (complex Rad1p-Rad10p) or TLS (Rev1p, Rev3p and Rad30p) proteins are associated with increased sensitivity to sodium valproate. The BER mutants (ogg1Δ, apn1Δ, rad27Δ, ntg1Δ and ntg2Δ) showed increased sensitivity to Cu(II) complexes. DNA damage induced by the complexes requires proteins from NER (Rad1p and Rad10p), TLS (Rev1p, Rev3p and Rad30p), PRR (Rad6 and Rad18p) and HR (Rad52p and Rad50p) for efficient repair. Therefore, Cu(II) complexes display enhanced cytotoxicity when compared to the sodium valproate and induce distinct DNA lesions, indicating a potential application as cytotoxic agents.
Asunto(s)
Cobre/farmacología , Reparación del ADN/efectos de los fármacos , Preparaciones Farmacéuticas/administración & dosificación , Fenantrolinas/farmacología , Saccharomyces cerevisiae/efectos de los fármacos , Ácido Valproico/farmacología , ADN/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , Ligandos , Mutación/efectos de los fármacos , Recombinación Genética/efectos de los fármacosRESUMEN
A series of selenylated-oxadiazoles were prepared and their interaction with DNA was investigated. The photophysical studies showed that all the selenylated compounds presented absorption between 270 and 329 nm, assigned to combined nâπ* and πâπ* transitions, and an intense blue emission (325-380 nm) with quantum yield in the range of Φ F = 0.1-0.4. DFT and TD-DFT calculations were also performed to study the likely geometry and the excited state of these compounds. Electrochemical studies revealed the ionization potential energies (-5.13 to -6.01 eV) and electron affinity energies (-2.25 to -2.83 eV), depending directly on the electronic effect (electron-donating or electron-withdrawing) of the substituent attached to the product. Finally, the UV-Vis DNA interaction experiments indicated that the compounds can interact with the DNA molecule due to intercalation, except for 3g (which interacted via electrostatic interaction). Plasmid cleavage assay presented positive results only for 3f that presented the strongest interaction results. These results made the tested selenylated-oxadiazoles as suitable structures for the development of drugs and the design of structurally-related therapeutics.
RESUMEN
In the search for new drugs, strategies such as bioisosterism have been evidenced, in which the modification of molecules is already known to be active. Thus, metal complexes of known drugs have been highlighted, with examples of significant improvements in therapeutic efficacy. In this way, this work aimed at the synthesis of new zinc complexes with nonsteroidal anti-inflammatory drugs (NSAIDs), as well as the chemical characterization and the previous toxicity by cytotoxicity with Artemia salina, and evaluating the ability of these compounds to interact with DNA. As result, two new zinc II ternary complexes containing the NSAIDs diclofenac (Diclof) and ibuprofen (Ibup) and nicotinamide neutral linker (Nic) were obtained by the two-step solvent metal-ligand complexation method. Molecular structures were determined by NMR, FTIR, HR-MS, UV-Vis and X-ray diffraction, which demonstrated that both complexes are binuclear systems of general formula [Zn2(R-COO-)4(Nic)2]. Plasmidial DNA breakdown capacities were evaluated by producing single and double breaks (DNA FII and FIII) from plasmid incubation with complex solutions in the concentration range 0 to 400 µmol·L-1 in experiments with the presence and absence of light. Both experiments did not show significant differences (P ≤ 0.05) in induced DNA cleavage activity between the maximum study concentrations (400 µmol·L-1) and the negative controls for both complexes. The types of complex 1 and 2 interactions with the secondary DNA structure were determined by titrating a CT-DNA solution with complex solutions and monitored by circular dichroism spectrometry. The results showed that both complexes interact with the grooves of the secondary structure of CT-DNA by electrostatic attraction, but without evidence of alteration in the primary structure. Acute toxicity tests against Artemia salina showed that both complexes did not produce lethality >10% of the population up to a maximum concentration of 1200 µg·mL-1 within an exposure interval of 24 h. Thus, two new compounds were synthesized, characterized and had their previous toxicities determined. These compounds are promising new drugs, with the next step being evaluations of their activity.
Asunto(s)
Artemia/crecimiento & desarrollo , Complejos de Coordinación/toxicidad , Inhibidores de la Ciclooxigenasa/toxicidad , Diclofenaco/química , Ibuprofeno/química , Niacinamida/química , Zinc/química , Animales , Antiinflamatorios no Esteroideos , Artemia/efectos de los fármacos , Complejos de Coordinación/química , Cristalografía por Rayos X , Inhibidores de la Ciclooxigenasa/química , División del ADN , Estructura Molecular , Pruebas de Toxicidad AgudaRESUMEN
BACKGROUND/AIM: Colorectal cancer is a common type of cancer with reported resistance to treatment, in most cases due to loss of function of apoptotic and cell-cycle proteins. Piperlongumine (PPLGM) is a natural alkaloid isolated from Piper species, with promising anti-cancer properties. This study investigated whether PPLGM is able to induce cell death in colorectal carcinoma HCT 116 cells expressing wild-type or deficient in Bax, p21 or p53. MATERIALS AND METHODS: PPLGM was extracted from roots of Piper tuberculatum. Cell viability was determined by reduction of 3-(4,5-dimethilthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and clonogenic assay. Cell death was evaluated by acridine orange/ethidium bromide staining and flow cytometry. Plasmid cleavage activity and circular dichroism DNA interaction were also analyzed. RESULTS: PPLGM induced selective cell death in all cell lines (IC50 range from 10.7 to 13.9 µM) with an increase in the number of late apoptotic cells and different profiles in cell-cycle distribution. Plasmid DNA analysis showed that PPLGM does not interact directly with DNA. CONCLUSION: This paper suggests that PPLGM may be a promising candidate in colorectal cancer therapy.
Asunto(s)
Neoplasias Colorrectales/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Dioxolanos/farmacología , Proteína p53 Supresora de Tumor/genética , Proteína X Asociada a bcl-2/genética , Apoptosis , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias Colorrectales/tratamiento farmacológico , Ensayos de Selección de Medicamentos Antitumorales , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Células HCT116 , HumanosRESUMEN
Investigations focused on the interactions of nanoparticles with lectins are relevant since it is well accepted that such proteins can be recognized by carbohydrates as parts of cell membranes. This can ultimately enhance the cellular uptake of the produced assemblies. In this framework, the physical interactions of phosphatidylcholine (PC) liposomes and the Bauhinia variegate lectin (BVL) are reported here. BVL-liposome interactions were characterized by a variety of techniques to understand the influence of BVL in the structural features, thermodynamic and spectroscopic properties of the hybrid material. The produced system is composed of 56% w/w lectin, and the scattering techniques show the presence of stable vesicular structures with a mean diameter DHâ¯â¼â¯100â¯nm. The FTIR and NMR results showed a strong lectin effect on the PC choline region, restricting the rotational motion of the lipid group. The BVL-liposome interaction promoted hardening of the protein as evidenced by circular dichroism spectroscopy. The photophysics results suggest higher rigidity of the system in the presence of BVL. The BVL may be present in the inner or outer polar surface of the liposomes. The system was shown to be relatively stable and therefore potentially useful for carbohydrate recognition of nanoparticles.
Asunto(s)
Bauhinia/química , Liposomas/química , Nanopartículas/química , Fosfatidilcolinas/química , Lectinas de Plantas/química , Tamaño de la Partícula , Conformación Proteica , Propiedades de SuperficieRESUMEN
Continuous increases in the rates of tumor diseases have highlighted the need for identification of novel and inexpensive antitumor agents from natural sources. In this study, we investigated the effects of enriched fraction from hydroalcoholic Brazilian red propolis extract against Hep-2 cancer cell line. Initially 201 fractions were arranged in 12 groups according to their chromatographic characteristics (A-L). After an in vitro cell viability screening, J and L were further selected as promising enriched fractions for this study. The chemical characterization was performed and Biochanin A, Formononetin, and Liquiritigenin compounds were quantified. Through MTT viability assay and morphological changes observed by Giemsa and DAPI staining, the results showed that red propolis inhibited cancer cells growth. Flow cytometry results indicated effects that were partly mediated through programmed cell death as confirmed by externalization of phosphatidylserine, DNA cleaved assay, increase at SUB G1-G0 phase in cell cycle analysis and loss of mitochondrial membrane potential. In conclusion, our results demonstrated that red propolis enriched fractions promoted apoptotic effects in human cancer cells through the mechanisms involving mitochondrial perturbation. Therefore, red propolis fractions contain candidate agents for adjuvant cancer treatment, which further studies should elucidate the comprehensive mechanistic pathways.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias/tratamiento farmacológico , Própolis/farmacología , Apoptosis/efectos de los fármacos , Brasil , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Flavanonas/farmacología , Fase G1/efectos de los fármacos , Genisteína/farmacología , Humanos , Isoflavonas/farmacología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Fase de Descanso del Ciclo Celular/efectos de los fármacosRESUMEN
Red ceramic industry in southern Brazil commonly uses wood biomass as furnace fuel generating great amounts of gas emissions and ash. To avoid their impact on atmospheric environment, wet scrubbing is currently being applied in several plants. However, the water leachate formed could be potentially toxic and not managed as a common water-based effluent, since the resulting wastewater could carry many toxic compounds derived from wood pyrolysis. There is a lack of studies regarding this kind of effluent obtained specifically and strictly from wooden-based biomass furnaces. Therefore, we conducted an evaluation of toxic and genotoxic potentials of this particular type of wet gas scrubber effluent. Physical-chemical analysis showed high contents of several contaminants, including phenols, sulphates and ammoniacal nitrogen, as well as the total and suspended solids. The effluent cause significant toxicity towards microcrustacean Artemia sp. (LC50 = 34.4%) and Daphnia magna (Toxicity Factor = 6 on average) and to higher plants (Lactuca sativa L. and Allium cepa L.) with acute and sub-acute effects in several parameters. Besides, using plasmid DNA, significant damage was observed in concentrations 12.5% and higher. In cellular DNA, concentrations starting from 12.5% and 6.25% showed significant increase in Damage Index (DI) and Damage Frequency (DF), respectively. The results altogether suggest that the effluent components, such phenols, produced by wood combustion can be volatilized, water scrubbed, resulting in a toxic and genotoxic effluent which could contaminate the environment.
Asunto(s)
Industria de la Construcción , Daño del ADN , Mutágenos/toxicidad , Aguas Residuales/química , Contaminantes Químicos del Agua/toxicidad , Madera/química , Animales , Artemia/efectos de los fármacos , Artemia/genética , Brasil , Cerámica , Materiales de Construcción , Daphnia/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Lactuca/efectos de los fármacos , Lactuca/genética , Cebollas/efectos de los fármacos , Cebollas/genéticaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ayurvedic and Chinese traditional medicine and tribal people use herbal preparations containing Piper nigrum fruits for the treatment of many health disorders like inflammation, fever, asthma and cancer. In Brazil, traditional maroon culture associates the spice Piper nigrum to health recovery and inflammation attenuation. AIMS OF THE STUDY: The aim of the current work was to evaluate the relationship between reactive oxygen species (ROS) overproduction, DNA fragmentation, cell cycle arrest and apoptosis induced by Piper nigrum ethanolic extract and its antitumor activity. METHODS: The plant was macerated in ethanol. Extract constitution was assessed by TLC, UV-vis and ESI-IT-MS/MS spectrometry. The cytotoxicity, proliferation and intracellular ROS generation was evaluated in MCF-7 cells. DNA damage effects were evaluated through intercalation into CT-DNA, plasmid DNA cleavage and oxidative damage in CT-DNA. Tumor growth inhibition, survival time increase, apoptosis, cell cycle arrest and oxidative stress were assessed in Ehrlich ascites carcinoma-bearing mice. RESULTS: Extraction yielded 64mg/g (36% piperine and 4.2% piperyline). Treatments caused DNA damage and reduced cell viability (EC50=27.1±2.0 and 80.5±6.6µg/ml in MCF-7 and HT-29 cells, respectively), inhibiting cell proliferation by 57% and increased ROS generation in MCF-7 cells (65%). Ehrlich carcinoma was inhibited by the extract, which caused reduction of tumor growth (60%), elevated survival time (76%), cell cycle arrest and induced apoptosis. The treatment with extract increased Bax and p53 and inhibited Bcl-xL and cyclin A expression. It also induced an oxidative stress in vivo verified as enhanced lipid peroxidation and carbonyl proteins content and increased activities of glutathione reductase, superoxide dismutase and catalase. GSH concentration was decreased in tumor tissue from mice. CONCLUSION: The ethanolic extract has cytotoxic and antiproliferative effect on MCF-7 cells and antitumor effect in vivo probably due to ROS overproduction that induced oxidative stress affecting key proteins involved in cell cycle arrest at G1/S and triggering apoptosis. Finally, the overall data from this study are well in line with the traditional claims for the antitumor effect of Piper nigrum fruits.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Carcinoma de Ehrlich/tratamiento farmacológico , Puntos de Control del Ciclo Celular/efectos de los fármacos , Daño del ADN , Etanol/química , Oxidantes/farmacología , Estrés Oxidativo/efectos de los fármacos , Piper nigrum/química , Piperidinas/farmacología , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/metabolismo , Solventes/química , Animales , Antineoplásicos Fitogénicos/aislamiento & purificación , Proteínas Reguladoras de la Apoptosis/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Carcinoma de Ehrlich/genética , Carcinoma de Ehrlich/metabolismo , Carcinoma de Ehrlich/patología , Proteínas de Ciclo Celular/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Células HT29 , Humanos , Peroxidación de Lípido/efectos de los fármacos , Células MCF-7 , Masculino , Ratones Endogámicos BALB C , Oxidantes/aislamiento & purificación , Fitoterapia , Piperidinas/aislamiento & purificación , Extractos Vegetales/aislamiento & purificación , Plantas Medicinales , Carbonilación Proteica/efectos de los fármacos , Factores de Tiempo , Carga Tumoral/efectos de los fármacos , Regulación hacia ArribaRESUMEN
Naphthoquinones interact with biological systems by generating reactive oxygen species (ROS) that can damage cancer cells. The cytotoxicity and the antitumor activity of 3acyl2phenylamino1,4naphthoquinones (DPB1DPB9) were evaluated in the MCF7 human breast cancer cell line and in male Ehrlich tumorbearing Balb/c mice. DPB4 was the most cytotoxic derivative against MCF7 cells (EC50 15 µM) and DPB6 was the least cytotoxic one (EC50 56 µM). The 1,4naphthoquinone derivatives were able to cause DNA damage and promote DNA fragmentation as shown by the plasmid DNA cleavage assay (FII form). In addition, 1,4naphthoquinone derivatives possibly interacted with DNA as intercalating agents, which was demonstrated by the changes caused in the fluorescence of the DNAethidium bromide complexes. Cell death of MCF7 cells induced by 3acyl2phenylamino1,4naphthoquinones was mostly due to apoptosis. The DNA fragmentation and subsequent apoptosis may be correlated to the redox potential of the 1,4naphthoquinone derivatives that, once present in the cell nucleus, led to the increased generation of ROS. Finally, certain 1,4naphthoquinone derivatives and particularly DPB4 significantly inhibited the growth of Ehrlich ascites tumors in mice (73%).
Asunto(s)
Antineoplásicos/toxicidad , Apoptosis/efectos de los fármacos , ADN/metabolismo , Sustancias Intercalantes/toxicidad , Naftoquinonas/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Animales , Antineoplásicos/química , Carcinoma de Ehrlich/tratamiento farmacológico , ADN/química , Daño del ADN/efectos de los fármacos , Humanos , Sustancias Intercalantes/química , Sustancias Intercalantes/uso terapéutico , Células MCF-7 , Masculino , Ratones , Ratones Endogámicos BALB C , Naftoquinonas/química , Naftoquinonas/uso terapéutico , Trasplante HeterólogoRESUMEN
CONTEXT: Croton celtidifolius Baill (Euphorbiaceae) is a tree found in the Atlantic Forest in Southern Brazil, where it is commonly known as "Sangue-de-Dragão". Its red latex is used traditionally for treating ulcers, diabetes and cancer. OBJECTIVE: To evaluate antitumor activities of Croton celtififolius latex in vitro and in vivo. MATERIAL AND METHODS: Phytochemical analyses were conducted using HPLC-DAD-MS. Cytotoxic, nuclease and pro-apoptotic properties were determined using the tetrazolium salt assay (MTT), plasmid DNA damage assay and ethidium bromide (EB)/acridine orange methods, respectively, and antitumor activity was determined in the Ehrlich ascites carcinoma (EAC) mouse model. RESULTS: Phytochemical studies indicated a high phenol content of flavonols (45.67 ± 0.24 and 18.01 ± 0.23 mg/mL of myricetin and quercetin, respectively) and flavan-3-ols (114.12 ± 1.84 and 1527.41 ± 16.42 mg/L of epicatechin and epigallocatechin, respectively) in latex. These compounds reduced MCF-7 and EAC cell viability in the MTT assay (IC50 = 169.0 ± 1.8 and 187.0 ± 2.2 µg/mL, respectively). Latex compounds caused significant DNA fragmentation and increased the number of apoptotic cells (negative control (NC), 12%; latex, 41%) as indicated by differential staining in the EB/acridine orange assay. The in vivo latex treatment at 3.12 mg/kg/day reduced the body weight by 7.57 ± 2.04 g and increased median survival time to 17.5 days when compared to the NC group (13.0 days). In addition, the highest latex concentration inhibited tumor growth by 56%. DISCUSSION AND CONCLUSION: These results agree with ethno-pharmacological reports showing cytotoxicity and antitumor activity of C. celtidifolius latex. The mechanism of antitumor action may be related to direct DNA fragmentation that reduces survival and induces apoptosis.
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Antineoplásicos Fitogénicos/farmacología , Croton/química , Flavonoles/farmacología , Extractos Vegetales/farmacología , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Brasil , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Carcinoma de Ehrlich/tratamiento farmacológico , Carcinoma de Ehrlich/patología , Supervivencia Celular/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , Femenino , Flavonoides/administración & dosificación , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Flavonoles/administración & dosificación , Flavonoles/aislamiento & purificación , Concentración 50 Inhibidora , Látex/administración & dosificación , Látex/aislamiento & purificación , Látex/farmacología , Células MCF-7 , Masculino , Medicina Tradicional , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Factores de TiempoRESUMEN
Pharmacological doses of ascorbate were evaluated for its ability to potentiate the toxicity of sodium orthovanadate (Na(3)VO(4)) in tumor cells. Cytotoxicity, inhibition of cell proliferation, generation of ROS and DNA fragmentation were assessed in T24 cells. Na(3)VO(4) was cytotoxic against T24 cells (EC(50)=5.8 µM at 24 h), but in the presence of ascorbate (100 µM) the EC(50) fell to 3.3 µM. Na(3)VO(4) plus ascorbate caused a strong inhibition of cell proliferation (up to 20%) and increased the generation of ROS (4-fold). Na(3)VO(4) did not directly cleave plasmid DNA, at this aspect no synergism was found occurring between Na(3)VO(4) and ascorbate once the resulting action of the combination was no greater than that of both substances administered separately. Cells from Ehrlich ascites carcinoma-bearing mice were used to determine the activity of antioxidant enzymes, the extent of the oxidative damage and the type of cell death. Na(3)VO(4) alone, or combined with ascorbate, increased catalase activity, but only Na(3)VO(4) plus ascorbate increased superoxide dismutase activity (up to 4-fold). Oxidative damage on proteins and lipids was higher due to the treatment done with Na(3)VO(4) plus ascorbate (2-3-fold). Ascorbate potentiated apoptosis in tumor cells from mice treated with Na(3)VO(4). The results indicate that pharmacological doses of ascorbate enhance the generation of ROS induced by Na(3)VO(4) in tumor cells causing inhibition of proliferation and apoptosis. Apoptosis induced by orthovanadate and ascorbate is closer related to inhibition on Bcl-xL and activation of Bax. Our data apparently rule out a mechanism of cell demise p53-dependent or related to Cdk2 impairment.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Ácido Ascórbico/farmacología , Proliferación Celular/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Vanadatos/farmacología , Animales , Línea Celular Tumoral , ADN/efectos de los fármacos , Fragmentación del ADN , Sinergismo Farmacológico , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Plásmidos/efectos de los fármacos , Proteína X Asociada a bcl-2/agonistas , Proteína bcl-X/antagonistas & inhibidoresRESUMEN
Landfill leachates are liquid effluents with elevated concentrations of chemical compounds that can cause serious environmental pollution. In the south of the state of Santa Catarina, Brazil, a sanitary landfill was installed that employs a system of anaerobic/facultative lagoons for the treatment of its leachate. The present work examined the toxic and genotoxic potential of untreated and treated landfill leachates using bioassays. The chemical, toxic, genotoxic and mutagenic properties of the untreated leachate and the treated leachate were determined. Examination of the chemical properties showed a marked decrease in parameters after treatment, as well as in toxicity towards all the organisms tested. The results of the comet assay demonstrated that both leachates showed genotoxicity in all of the organisms tested, indicating the persistence of genotoxic substances even after treatment. A significant decrease in micronucleated cells was detected in Geophagus brasiliensis exposed to the treated leachate compared to untreated.
RESUMEN
Many biological properties have been attributed to ruthenium complexes including anti-tumor activity and the attenuation of reperfusion damage and infarct size. In this work, we characterize the antioxidant activity of trans-[RuCl2(nic)4] where nic is 3-pyridinecarboxylic acid and trans-[RuCl2(i-nic)4] where i-nic is 4-pyridinecarboxylic acid by (i) evaluation of total antioxidant potential (TRAP); (ii) prevention of DNA damage induced by hydrogen peroxide using the alkaline comet assay; and (iii) the prevention of lipid peroxidation and cell death induced by iron in liver slices. Our results suggest that nic has stronger antioxidant potential when compared to the i-nic. Higher doses (above 200 microM) of these compounds gave genotoxic effects, but the antioxidant potential could be obtained with the use lower doses (0.1-10 microM).