RESUMO
Bisphosphonates are the most commonly prescribed drugs for the treatment of osteoporosis and other bone illnesses. Some of them have also shown antiparasitic activity. In search of improving the pharmacological profile of commercial bisphosphonates, our group had previously developed first row transition metal complexes with N-containing bisphosphonates (NBPs). In this work, we extended our studies to heteroleptic palladium-NBP complexes including DNA intercalating polypyridyl co-ligands (NN) with the aim of obtaining potential multi-target species. Complexes of the formula [Pd(NBP)2(NN)]·2NaCl·xH2O with NBP = alendronate (ale) or pamidronate (pam) and NN = 1,10 phenanthroline (phen) or 2,2'-bipyridine (bpy) were synthesized and fully characterized. All the obtained compounds were much more active in vitro against T. cruzi (amastigote form) than the corresponding NBP ligands. In addition, complexes were nontoxic to mammalian cells up to 50-100 µM. Compounds with phen as ligand were 15 times more active than their bpy analogous. Related to the potential mechanism of action, all complexes were potent inhibitors of two parasitic enzymes of the isoprenoid biosynthetic pathway. No correlation between the anti-T. cruzi activity and the enzymatic inhibition results was observed. On the contrary, the high antiparasitic activity of phen-containing complexes could be related to their ability to interact with DNA in an intercalative-like mode. These rationally designed compounds are good candidates for further studies and good leaders for future drug developments. Four new palladium heteroleptic complexes with N-containing commercial bisphosphonates and DNA intercalating polypyridyl co-ligands were synthesized and fully characterized. All complexes displayed high anti-T. cruzi activity which could be related to the inhibition of the parasitic farnesyl diphosphate synthase enzyme but mainly to their ability to interact DNA.
Assuntos
Complexos de Coordenação/farmacologia , Difosfonatos/farmacologia , Paládio/farmacologia , Tripanossomicidas/farmacologia , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Difosfonatos/química , Estrutura Molecular , Paládio/química , Testes de Sensibilidade Parasitária , Tripanossomicidas/síntese química , Tripanossomicidas/química , Trypanosoma cruzi/efeitos dos fármacosRESUMO
Osteosarcoma (OS) is the most common primary tumor of bone, occurring predominantly in the second decade of life. High-dose cytotoxic chemotherapy and surgical resection have improved prognosis, with long-term survival for patients with localized disease. Vanadium is an ultra-trace element that after being absorbed accumulates in bone. Besides, vanadium compounds have been studied during recent years to be considered as representative of a new class of non-platinum antitumor agents. Moreover, flavonoids are a wide family of polyphenolic compounds that display many interesting biological effects. Since coordination of ligands to metals can improve the pharmacological properties, we report herein, for the first time, the in vitro and in vivo effects of an oxidovanadium(IV) complex with the flavonoid chrysin on the new 3D human osteosarcoma and xenograft osteosarcoma mice models. The pharmacological results show that VOchrys inhibited the cell viability affecting the shape and volume of the spheroids and VOchrys suppressed MG-63 tumor growth in the nude mice without inducing toxicity and side effects. As a whole, the results presented herein demonstrate that the antitumor action of the complex was very promissory on human osteosarcoma models, whereby suggesting that VOchrys is a potentially good candidate for future use in alternative antitumor treatments.
Assuntos
Neoplasias Ósseas/tratamento farmacológico , Complexos de Coordenação/farmacologia , Flavonoides/farmacologia , Osteossarcoma/tratamento farmacológico , Esferoides Celulares/efeitos dos fármacos , Vanádio/farmacologia , Animais , Neoplasias Ósseas/patologia , Técnicas de Cultura de Células/métodos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/química , Feminino , Flavonoides/química , Humanos , Masculino , Camundongos Nus , Microscopia de Contraste de Fase , Estrutura Molecular , Osteossarcoma/patologia , Esferoides Celulares/patologia , Fatores de Tempo , Resultado do Tratamento , Vanádio/química , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The α6ß4 integrin is composed of the α6 and ß4 subunits that are encoded by the ITGα6 and the ITGß4 genes, respectively. The α6ß4 main function is to intervene in lamination and epithelia integrity maintenance by cell-matrix interactions. This integrin appears to have importance in breast cancer malignancy, as well as other epithelial carcinomas. The aim of this work was to investigate the potential role of ITGα6 (A380T) and ITGß4 (R1281W) genetic variations in breast cancer susceptibility, in a female population from the northeast region of Argentina (Misiones). We performed a case-control study of 85 breast cancer patients and 113 cancer-free controls. Genotyping was performed by RFLP-PCR. For ITGα6 (A380T) single nucleotide polymorphism, a high frequency of heterozygous genotype GA in cases compared to controls was observed, achieving values of 48% and 49%, respectively. No association between the A380T SNP and breast cancer development was found (Odds Ratio = 0.92; 95% Confidence Interval = 0.52-1.63; p = 0.884). In conclusion, we did not find evidence of an association between A380T (ITGα6) and the risk of developing breast cancer. The results represent the first report of these genetic variations in breast cancer; therefore, they are an important contribution to the literature.
Assuntos
Neoplasias da Mama/genética , Integrina alfa6/genética , Integrina beta4/genética , Polimorfismo de Nucleotídeo Único , Adulto , Idoso , Argentina/epidemiologia , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/patologia , Estudos de Casos e Controles , Feminino , Frequência do Gene , Variação Genética , Genótipo , Humanos , Pessoa de Meia-Idade , Razão de Chances , RiscoRESUMO
Vanadium is a transition metal that, being ubiquitously distributed in soil, crude oil, water and air, also found roles in biological systems and is an essential element in most living beings. There are also several groups of organisms which accumulate vanadium, employing it in their biological processes. Vanadium being a biological relevant element, it is not surprising that many vanadium based therapeutic drugs have been proposed for the treatment of several types of diseases. Namely, vanadium compounds, in particular organic derivatives, have been proposed for the treatment of diabetes, of cancer and of diseases caused by parasites. In this work we review the medicinal applications proposed for vanadium compounds with particular emphasis on the more recent publications. In cells, partly due to the similarity of vanadate and phosphate, vanadium compounds activate numerous signaling pathways and transcription factors; this by itself potentiates application of vanadium-based therapeutics. Nevertheless, this non-specific bio-activity may also introduce several deleterious side effects as in addition, due to Fenton's type reactions or of the reaction with atmospheric O2, VCs may also generate reactive oxygen species, thereby introducing oxidative stress with consequences presently not well evaluated, particularly for long-term administration of vanadium to humans. Notwithstanding, the potential of vanadium compounds to treat type 2 diabetes is still an open question and therapies using vanadium compounds for e.g. antitumor and anti-parasitic related diseases remain promising.
RESUMO
A new complex of the oxovanadium(IV) cation with the flavolignan silibinin has been synthesized and characterized. Vanadium compounds show interesting biological and pharmacological properties and some of them display antitumoral actions. Flavonoids are part of a larger group of antioxidant compounds called polyphenols which may inhibit the proliferation and growth of cancer cells. The antioxidant and antitumoral effects of silibinin and its oxovanadium(IV) complex were investigated. Silibinin acted as a very strong antioxidant and its complexation with oxovanadium(IV) improved this behavior. Besides, the generation of reactive oxygen species (ROS) by this compound was favored in tumoral (UMR106) cells and correlated with the deleterious behavior in the proliferation of this cell line. Conversely, silibinin did not exert any effect on the proliferation of normal osteoblasts (MC3T3E1). The cytotoxic action and ROS generation of the oxovanadium(IV) complex was more effective in tumoral cells. This behavior was not consistent with cleaving DNA of plasmid DNA pA1 because no significant cleaving activity was observed in both cases. These results suggest that the main deleterious mechanisms may take place through cytotoxic effects more than genotoxic actions. A comparison with our own findings on the behavior of other flavonoids and their vanadyl(IV) complex has also been performed.
Assuntos
Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Clivagem do DNA/efeitos dos fármacos , Compostos Organometálicos/farmacologia , Silimarina/farmacologia , Vanadatos/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antioxidantes/síntese química , Antioxidantes/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Espectroscopia de Ressonância de Spin Eletrônica , Estrutura Molecular , Compostos Organometálicos/síntese química , Compostos Organometálicos/química , Plasmídeos/efeitos dos fármacos , Ratos , Espécies Reativas de Oxigênio/metabolismo , Silibina , Silimarina/química , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Relação Estrutura-Atividade , Vanadatos/químicaRESUMO
The complex of vanadyl(IV) cation with oxodiacetate, VO(oda) caused an inhibitory effect on the proliferation of the human colon adenocarcinoma cell line Caco-2 in the range of 25-100 µM (P < 0.001). This inhibition was partially reversed by scavengers of free radicals. The difference in cell proliferation in the presence and the absence of scavengers was statistically significant in the range of 50-100 µM (P < 0.05). VO(oda) altered lysosomal and mitochondria metabolisms (neutral red and MTT bioassays) in a dose-response manner from 10 µM (P < 0.001). Morphological studies showed important transformations that correlated with the disassembly of actin filaments and a decrease in the number of cells in a dose response manner. Moreover, VO(oda) caused statistically significant genotoxic effects on Caco-2 cells in the low range of concentration (5-25 µM) (Comet assay). Increment in the oxidative stress and a decrease in the GSH level are the main cytotoxic mechanisms of VO(oda). These effects were partially reversed by scavengers of free radicals in the range of 50-100 µM (P < 0.05). Besides, VO(oda) interacted with plasmidic DNA causing single and double strand cleavage, probably through the action of free radical species. Altogether, these results suggest that VO(oda) is a good candidate to be evaluated for alternative therapeutics in cancer treatment.
Assuntos
Acetatos/toxicidade , Acetatos/uso terapêutico , Células CACO-2/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Vanadatos/toxicidade , Vanadatos/uso terapêutico , Acetatos/química , Actinas/metabolismo , Animais , Células CACO-2/citologia , Proliferação de Células/efeitos dos fármacos , Ensaio Cometa , Citoesqueleto/efeitos dos fármacos , Dano ao DNA , Relação Dose-Resposta a Droga , Glutationa/metabolismo , Humanos , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Vanadatos/químicaRESUMO
The new complex [VO(chrysin)(2)EtOH](2) (VOchrys) has been synthesized and thoroughly characterized. Fourier transform IR, UV-vis, diffuse reflectance, and EPR spectroscopies as well as elemental analysis and thermal measurements were performed. In solution, different species could be detected by EPR spectroscopy as a function of the ligand-to-metal ratio. The stoichiometry of the chelate complex formed at pH 5 was also determined by spectrophotometric titrations. Since flavonoids are natural antioxidant compounds, the antioxidant capacity of chrysin and its vanadyl(IV) complex was investigated using different radicals. Chrysin and its complex were not able to diminish the level of superoxide and 1,1-diphenyl-2-picrylhydrazyl radicals to a great extent. In contrast, they were strong scavengers for 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt radical cations and OH. radicals with a greater potency for VOchrys. Taking into account their selective antioxidant properties, we investigated the bioactivity of these compounds in two osteoblast-like cells in culture. Chrysin and VOchrys caused an inhibition of cell proliferation in MC3T3E1 normal osteoblasts and UMR106 tumor cells in a dose-response manner, with a greater effect in the latter cell line. The generation of reactive oxygen species (ROS) was evaluated in both cell lines and a correlation could be established between the antiproliferative effects of chrysin and the increase in the ROS levels. The complex did not generate types of ROS that can be detected by the dihydrorhodamine 123 technique so the antiproliferative effect may be attributed to the formation of other radicals such as superoxide, which is not detected by this probe. The morphological alterations were in agreement with these changes.
Assuntos
Antineoplásicos/farmacologia , Flavonoides/química , Sequestradores de Radicais Livres/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Vanadatos/química , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sequestradores de Radicais Livres/síntese química , Sequestradores de Radicais Livres/química , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Camundongos , Osteoblastos/citologia , Ratos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Vanadium is a trace element widely distributed in the environment. In vertebrates it is mainly stored in bone tissue. The unique cellular environment in the bone and the variety of interactions that mediate cancer metastasis determine that certain types of cancer, such as breast and prostate cancer, preferentially metastize in the skeleton. Since this effect usually signifies serious morbidity and grave prognosis there is an increasing interest in the development of new treatments for this pathology. The present work shows that vanadium complexes can inhibit some parameters related to cancer metastasis such as cell adhesion, migration and clonogenicity. We have also investigated the role of protein kinase A in these processes.
Assuntos
Metástase Neoplásica/prevenção & controle , Osteossarcoma/tratamento farmacológico , Oligoelementos/farmacologia , Vanádio/farmacologia , Animais , Aspirina/química , Aspirina/farmacologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Proteínas Quinases Dependentes de AMP Cíclico/efeitos dos fármacos , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Estabilidade de Medicamentos , Glucose/química , Glucose/farmacologia , Ratos , Oligoelementos/química , Trealose/química , Trealose/farmacologia , Vanádio/químicaRESUMO
A new copper(II) complex of santonic acid [Cu(2)(sant)(4)(H(2)O)(2)].2(1/2)H(2)O has been prepared and characterized by electronic, vibrational, EPR spectral studies, and stability determinations in solution. The presence of two antiferrromagnetically coupled copper centers in the solid state was detected by EPR. The dinuclear Cu(II) complex crystallizes in the tetragonal P4(3)2(1)2 space group, with a=b=14.498(3), c=64.07(1)A. Biological studies indicate that the complex displays interesting potential antitumoral actions.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Hidrocarbonetos Aromáticos com Pontes/síntese química , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Cobre/química , Animais , Antineoplásicos/química , Hidrocarbonetos Aromáticos com Pontes/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Cristalografia por Raios X , Espectroscopia de Ressonância de Spin Eletrônica , Humanos , Camundongos , Modelos Moleculares , Estrutura Molecular , Ratos , Espectrofotometria Infravermelho , Análise Espectral Raman , Relação Estrutura-Atividade , VibraçãoRESUMO
In this study we report the synthesis, characterization and a thorough biological evaluation of twelve organoruthenium-8-hydroxyquinolinato (Ru-hq) complexes. The chosen hqH ligands bear various halogen atoms in different positions which enables to study effect of the substituents on physico-chemical and biological properties. The determined crystal structures of novel complexes expectedly show the cymene ring, a bidentately coordinated deprotonated hq and a halide ligand (chlorido or iodido) coordinated to the ruthenium central ion. In previous studies the anticancer potential of organoruthenium complex with 8-hydroxyquinoline ligand clioquinol was well established and we have decided to perform an extended biological evaluation (antibacterial and antitumor activity) of the whole series of halo-substituted analogs. Beside the cytotoxic potential of studied compounds also the effect of two selected complexes (9 and 10) on apoptosis induction in MG-63 and A549 cells was also studied via externalization of phosphatidylserine at the outer plasma membrane leaflet. Both selected complexes that gave best preliminary cytotoxicity results contain bromo substituted hq ligands. Apoptosis induction results are in agreement with the cell viability assays suggesting the higher and more selective anticancer activity of complex 10 in comparison to complex 9 on MG-63 cells.
Assuntos
Antibacterianos , Antineoplásicos , Apoptose/efeitos dos fármacos , Neoplasias , Compostos Organometálicos , Oxiquinolina , Rutênio , Células A549 , Antibacterianos/síntese química , Antibacterianos/química , Antibacterianos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Neoplasias/patologia , Compostos Organometálicos/síntese química , Compostos Organometálicos/química , Compostos Organometálicos/farmacologia , Oxiquinolina/química , Oxiquinolina/farmacologia , Rutênio/química , Rutênio/farmacologiaRESUMO
Cancer is a group of diseases involving abnormal cell growth. The cells grow uncontrollably with the potential to invade and spread to other parts of the body. This disease is one of the principal death causes in the world, thus becoming a significant topic of scientific research. On the other hand, transition metals play a fundamental role in different living systems. In particular, Metallodrugs represent new and powerful tools for diverse therapeutic applications. To date, various metallodrugs display interesting biological activities for chemotherapy. In this field, cisplatin was the first inorganic compound with high relevance in cancer treatment. This compound was a leader agent in clinical use. Toxicity and resistance problems trigger the development of other platinum drugs with better clinical perspective and also raise the scientific interest for the putative antitumor properties of V, Ru and Cu compounds. Several scientific articles show that complexes of these metals are the new metal-based drugs used in the treatment of several cancers, such us, lung, colon, breast, bladder, etc. In this review we recapitulate current information and new advances on antitumor in vitro effects of several organic and inorganic compounds derived from copper, ruthenium and vanadium. These metal derived compounds targeting DNA or cell proteins involved in cell signaling pathways related to cancer. The mechanisms of cell death of these metallodrugs have also been comprehensibly reviewed. The knowledge of these mechanisms of death and the relationship between chemical structure and biological activity may be useful for the design of new metal-based drugs with promising pharmacologic applications as anticancer agents.
Assuntos
Antineoplásicos/farmacologia , Complexos de Coordenação/farmacologia , Cobre/química , Rutênio/química , Vanádio/química , Animais , Apoptose/efeitos dos fármacos , Portadores de Fármacos , Ácidos Graxos Monoinsaturados/química , Humanos , Nanopartículas/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Compostos de Amônio Quaternário/químicaRESUMO
Vanadium compounds were studied in recent years by considering them as a representative of a new class of non-platinum metal anticancer drugs. However, a few challenges still remain in the discovery of new molecular targets of these new metallodrugs. Studies on cell signaling pathways related to vanadium compounds have scarcely been reported and so far this information is highly critical for identifying novel targets that play a key role in the antitumor actions of vanadium complexes. This research deals with the alterations in the intracellular signaling pathways promoted by an oxovanadium(iv) complex with the clioquinol (5-chloro-7-iodo-8-quinolinol), VO(CQ)2, on a human osteosarcoma cell line (MG-63). Herein are reported, for the first time, the antitumor properties of VO(CQ)2 and the relative abundance of 224 proteins (which are involved in most of the common intracellular pathways) to identify novel targets of the studied complex. Besides, full-length human recombinant AKT1 kinase was produced by using an IVTT system to evaluate the variation of relative tyrosin-phosphorylation levels caused by this compound. The results of the differential protein expression levels reveal several up-regulated proteins such as CASP3, CASP6, CASP7, CASP10, CASP11, Bcl-x, DAPK and down-regulated ones, such as PKB/AKT, DIABLO, among others. Moreover, cell signaling pathways involved in several altered pathways related to the PKC and AP2 family have been identified in both treatments (2.5 and 10 µM) suggesting the crucial antitumoral role of VO(CQ)2. Finally, it has been demonstrated that this compound (10 µM, 6 h) triggers a decrease of 2-fold in in situ AKT1 expression.
Assuntos
Antineoplásicos/farmacologia , Clioquinol/farmacologia , Osteossarcoma/patologia , Transdução de Sinais/efeitos dos fármacos , Compostos de Vanádio/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Clioquinol/síntese química , Clioquinol/química , Perfilação da Expressão Gênica , Humanos , Reprodutibilidade dos Testes , Compostos de Vanádio/síntese química , Compostos de Vanádio/químicaRESUMO
BACKGROUND: Copper has shown to be useful in disorders with an inflammation origin such as cancer [1-3]. It has previously shown that Casiopeínas® interact with DNA and promote the disruption by a mechanism related to the increase in the level of free radicals [4-6] which confers antineoplastic potential. Objetive: The aim of the present work was to study the antitumor effects of a series of Cu(II) complexes with saccharinate (sac) and glutamate (gln): [Cu(sac)2(H2O)4].2H2O (Cu-sac), [Cu(gln)2] (Cu-gln) and Na2[Cu(sac)2 (gln)2].H2O (Cu-sac-gln). METHODS: We have investigated the action of these compounds on cell viability on human osteosarcoma cells MG-63. In particular, we pay special attention to the cyto and genotoxicity actions of these complexes and to the association to oxidative stress. RESULTS: The three complexes: Cu-sac, Cu-gln and Cu-sac-gln caused a decline in cell viability. The half-maximal inhibitory concentration in MG-63 cells for Cu-sac-gln is 170 µM, showing the strongest antiproliferative effect. Moreover, only Cu-sac-gln caused a decrease of the mitochondrial activity from 100 µM. Our results indicate that the copper(II) complexes studied here produce DNA damage and suggest that the rise of reactive oxygen species (ROS) is the central mechanism action. Genotoxicity studied by the Cytokinesis-block micronucleus (MN) assay and the Single cell gel electrophoresis (comet assay) could be observed in MG-63 cells treated with Cu-sac-gln from 100 and 50 µM, respectively. Cu-sac and Cu-gln also induced DNA damage; however their effect was definitively weaker. The generation of reactive oxygen species increased from 50 µM of Cu-sac-gln and Cu-sac and only from 250 µM of Cugln, as well as a reduction of the GSH/GSSG ratio from 50 µM. When cells were treated with several concentrations of the complexes in addition to a combination of 50 µM of vitamin C plus 50 µM of vitamin E, a total recovery in cell survival was obtained for Cu-gln in the whole range of tested concentrations while only a partial viability recovery was obtained from 250 µM of Cu-sac and Cu-sac-gln. CONCLUSION: Overall, our results point to a differential cyto- and genotoxicity of the three copper(II) complexes and demonstrate that the complexation with both ligands confers the most potent antitumor action in human osteosarcoma cells.
Assuntos
Antineoplásicos/farmacologia , Cobre/farmacologia , Glutamina/farmacologia , Compostos Organometálicos/farmacologia , Osteossarcoma/tratamento farmacológico , Sacarina/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cobre/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Glutamina/química , Humanos , Camundongos , Estrutura Molecular , Compostos Organometálicos/síntese química , Compostos Organometálicos/química , Osteossarcoma/patologia , Sacarina/análogos & derivados , Sacarina/química , Relação Estrutura-AtividadeRESUMO
Oxovanadium(IV) complexes of the polyalcohols sorbitol, galactitol, and mannitol, of stoichiometry Na(2)[VO(L)(2)].H(2)O, were obtained from aqueous alkaline solutions. They were characterized by elemental analysis, infrared and UV-vis spectroscopies, thermoanalytical (thermogravimetric and differential thermal analysis) data, and magnetic susceptibility measurements. The biological activities of the complexes on the proliferation, differentiation, and glucose consumption were tested on osteoblast-like cells (MC3T3E1 osteoblastic mouse calvaria-derived cells and UMR106 rat osteosarcoma-derived cells) in culture. The three complexes exerted a biphasic effect on cell proliferation, being slight stimulating agents at low concentrations and inhibitory in the range of 25-100 microM. All the complexes inhibited cell differentiation in tumor osteoblasts. Their effects on glucose consumption were also discussed. The free ligands did not show any effect on the studied biological parameters.
Assuntos
Álcoois/química , Compostos de Vanádio/química , Compostos de Vanádio/síntese química , Animais , Proliferação de Células , Células Cultivadas , Temperatura Alta , Raios Infravermelhos , Camundongos , Estrutura Molecular , Células Tumorais CultivadasRESUMO
Vanadium complexes were studied during recent years and considered as a representative of a new class of non-platinum metal antitumor agents in combination with their low toxicity. However, a few challenges still remain in the discovery of new molecular targets for these novel metal-based drugs. The study of cell signaling pathways related to vanadium drugs, which is highly critical for identifying specific targets that play an important role in the antitumor activity of vanadium compounds, is scarce. This research deals with the alterations in intracellular signaling pathways promoted by an oxovanadium(iv) complex with the flavonoid chrysin [VO(chrysin)2EtOH]2 (VOChrys) in a human osteosarcoma cell line (MG-63). Herein we report for the first time the effect of [VO(chrysin)2EtOH]2 on the relative abundance of 224 proteins, which are involved in the most common intracellular pathways. Besides, full-length human recombinant (FAK and AKT1) kinases are produced using an in situ IVTT system and then we have evaluated the variation of relative tyrosine-phosphorylation levels caused by the [VO(chrysin)2EtOH]2 compound. The results of the differential protein expression levels reveal that several proteins such as PKB/AKT, PAK, DAPK, Cdk 4, 6 and 7, FADD, AP2, NAK, and JNK, among others, were altered. Moreover, cell signaling pathways related to the PTK2B, FAK, PKC families suggests an important role associated with the antitumor activity of [VO(chrysin)2EtOH]2 was demonstrated. Finally, the effect of this compound on in situ expressed FAK and AKT1 is validated by determining the phosphorylation level, which decreased in the former and increased in the latter.
Assuntos
Antineoplásicos/farmacologia , Neoplasias Ósseas/patologia , Flavonoides/química , Compostos Organometálicos/farmacologia , Osteossarcoma/patologia , Vanadatos/química , Antineoplásicos/química , Neoplasias Ósseas/tratamento farmacológico , Sobrevivência Celular , Humanos , Estrutura Molecular , Compostos Organometálicos/química , Osteossarcoma/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
Bone homeostasis is the result of a tight balance between bone resorption and bone formation where macrophage activation is believed to contribute to bone resorption. We have previously shown that a vanadyl(IV)-aspirin complex (VOAspi) regulates cell proliferation and differentiation of osteoblasts in culture. In this study, we assessed VOAspi and VO effects and their possible mechanism of action on a mouse macrophage cell line RAW 264.7. Both vanadium compounds inhibited cell proliferation in a dose-dependent manner. Nifedipine completely reversed the VOAspi-induced macrophage cytotoxicity, while it could not block the effect of VO. VOAspi also stimulated nitric oxide (NO) production, the oxidation of dihydrorhodamine 123 (DHR-123) and enhanced the expression of both constitutive and inducible isoforms of nitric oxide syntases (NOS). All these effects were abolished by nifedipine. Altogether our finding give evidence that VOAspi-induced macrophage cytotoxicity is dependent on L-type calcium channel and the generation of NO though the induction of eNOS and iNOS. Contrary, the parent compound VO exerted a cytotoxic effect by mechanisms independent of a calcium entry and the NO/NOS activation.
Assuntos
Aspirina/análogos & derivados , Aspirina/toxicidade , Canais de Cálcio Tipo L/metabolismo , Proliferação de Células/efeitos dos fármacos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Óxido Nítrico/biossíntese , Compostos de Vanádio/toxicidade , Animais , Western Blotting , Linhagem Celular , Relação Dose-Resposta a Droga , Macrófagos/metabolismo , Camundongos , Microscopia de Fluorescência , Óxido Nítrico Sintase/biossíntese , Espécies Reativas de Oxigênio/metabolismoRESUMO
Oxovanadium (IV) complexes of the cyclic polyols conduritol C (cond) and myo-inositol (inos) of stoichiometry Na(2)[VO(cond)(2)].2H(2)O and Na(2)[VO(inos)(2)].H(2)O were obtained in aqueous alkaline solutions. They were characterized by infrared and UV-Vis spectroscopies, thermoanalytical (thermogravimetric and differential thermal analysis) data and magnetic susceptibility measurements. The biological activities of the complexes on the proliferation, differentiation and glucose consumption were tested on osteoblast-like cells in culture. Conduritol C and myo-inositol did not produce any effect on these parameters. Normal and tumoral cell proliferation was inhibited about (ca.40-60%) by the two oxovanadium (IV) complexes in concentrations as low as 100microM. The complexes were also inhibitory on cell differentiation (ca. 70-80%) while they stimulate glucose consumption. Comparisons of these effects with those of the oxovanadium (IV) cation, under the same experimental conditions, were also performed.
Assuntos
Compostos Organometálicos/síntese química , Compostos Organometálicos/farmacologia , Vanadatos/síntese química , Vanadatos/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Cicloexanóis/síntese química , Cicloexanóis/química , Cicloexanóis/farmacologia , Cicloexenos , Glucose , Inositol/síntese química , Inositol/química , Inositol/farmacologia , Camundongos , Estrutura Molecular , Compostos Organometálicos/química , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Ratos , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de Fourier , Termodinâmica , Vanadatos/químicaRESUMO
The oxidovanadium(IV) complex of oxodiacetic acid (H2ODA) and dppz (dipyrido[3,2-a:2',3'-c] phenazine) of stoichiometry [VO(ODA)(dppz)]·3H2O could be synthesized for the first time by reaction between [VO(ODA)(H2O)2] and dppz. It was characterized by infrared and electronic spectroscopies. Its optimized molecular structure was obtained by DFT calculations, as it was impossible to grow single crystals adequate for crystallographic studies. The antitumor action of the complex on MG-63 human osteosarcoma cell line was also investigated. It was found that it caused a concentration-related inhibitory effect in the concentration range between 5 and 25 µM and diminished the cell viability ca. 45% in the range from 25 to 100 µM, without dose/response effects in this range. These biological effects are, in general, similar to those previously reported for the related [VO(ODA)(ophen)]·1.5H2O complex.
Assuntos
Ácido Acético/química , Compostos Organometálicos/química , Fenazinas/química , Vanadatos/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Compostos Organometálicos/síntese química , Compostos Organometálicos/farmacologia , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The adhesion of osteoblasts to bone extracellular matrix, of which type-I collagen constitutes >85%, can modulate diverse aspects of their physiology such as growth, differentiation and mineralisation. In this study we examined the adhesion of UMR106 rat osteoblast-like cells either to a control (Col) or advanced-glycation-endproduct-modified (AGEs-Col) type I collagen matrix. We investigated the possible role of different integrin receptors in osteoblastic adhesion, by co-incubating these cells either with beta-peptide (conserved sequence 113-125 of the beta subunit of integrins) or with two other peptides, RGD (Arg-Gly-Asp) and DGEA (Asp-Gly-Glu-Ala), which are recognition sequences for the alpha-subunits of alpha(1,5)beta(1) and alpha(2)beta(1) integrins. Collagen glycation inhibited the adhesion of UMR106 osteoblasts to the matrix (40% reduction versus Col, P > 0.001). beta-Peptide showed a dose- and glycation-dependent inhibitory effect on adhesion, and at a concentration of 100 microM decreased the attachment of UMR106 cells to both matrices (42% to Col, P<0.001and 25% to AGEs-Col, P<0.01). The synthetic peptides RGD (1mM) and DGEA (5mM) inhibited the attachment of UMR106 cells to Col (30 and 20%, P > 0.01 and P< 0.001, respectively), but not to AGEs-Col. beta-Peptide induced an increase in UMR106 cell clumping and a decrease in cellular spreading, while DGEA increased spreading with cellular extensions in multiple directions. These results indicate that both alpha and beta integrin subunits participate in osteoblastic attachment to type-I collagen, probably through the alpha(1,5)beta(1) and alpha(2)beta(1) integrins. AGEs-modification of type-I collagen impairs the integrin-mediated adhesion of osteoblastic cells to the matrix, and could thus contribute to the pathogenesis of diabetic osteopenia.
Assuntos
Colágeno Tipo I/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Integrinas/antagonistas & inibidores , Osteoblastos/metabolismo , Animais , Matriz Óssea/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Glicosilação , Cadeias alfa de Integrinas/metabolismo , Cadeias beta de Integrinas/metabolismo , Integrinas/fisiologia , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteogênese , Peptídeos/metabolismo , Peptídeos/farmacologia , RatosRESUMO
BACKGROUND: Vanadium derivatives have been reported to display different biological effects, and in particular antineoplastic activity has been demonstrated in both in vivo and in vitro studies. PURPOSE. To study the effect of two new organic vanadyl(IV) complexes (one with glucose, GluVO, and the other with naproxen, NapVO) in osteosarcoma cells. METHODS: UMR106 osteosarcoma cells and, for comparison, nontransformed MC3T3E1 osteoblasts were used. Proliferation and differentiation were assessed using the crystal violet assay and ALP specific activity, respectively. Morphological alterations were assessed by light microscopy. Lipid peroxidation was evaluated in terms of production of thiobarbituric acid-reactive substances (TBARS) and apoptosis was measured using annexin V. Extracellular regulated kinase (Erk) activation was investigated by Western blotting. RESULTS: Vanadium complexes caused morphological alterations and they strongly inhibited UMR106 cell proliferation and differentiation. In contrast, in MC3T3E1 cells, these vanadium derivatives had a relatively weak action. In UMR106 tumoral cells there was a significant increase in TBARS production. Both vanadium complexes induced apoptosis and activation of Erk. PD98059, an inhibitor of Erk phosphorylation, did not block the vanadium-induced antitumoral action. However, the antioxidants vitamins C and E abrogated the apoptosis and TBARS production induced by the vanadium complexes. CONCLUSIONS: GluVO and NapVO exerted an antitumoral effect in UM106 osteosarcoma cells. They inhibited cell proliferation and differentiation. While the Erk cascade seems not to be directly related to the bioactivity of these vanadium derivatives, the action of both vanadium complexes with organic ligands may be mediated by apoptosis and oxidative stress.