RESUMEN
Osteosarcoma (OS) is a primary malignant bone tumor that has an abnormal expression of oncogenesis and tumor suppressors and causes dysregulation of various signaling pathways. Thus, novel therapeutic strategies for OS are needed to overcome the resistance of traditional treatments. This study evaluated the cytotoxic and anticancer effects of the association between menadione (MEN) and protocatechuic acid (PCA) in murine OS cells (UMR-106). The concentrations were 3.12 µM of isolated MEN, 500 µM of isolated PCA, and their associations. We performed cell viability assays, morphology modification analysis, cell migration by the wound-healing method, apoptosis by flow cytometry, reactive oxygen species (ROS) production, gene expression of NOX by RT-qPCR, and degradation of MMP-2 and 9 by zymography. Our results showed that the association of MEN+PCA was more effective in OS cells than the compounds alone. The association decreased cell viability, delayed cell migration, and decreased the expression of NOX-2 and ROS. In addition, the MEN+PCA association induced a slight increase in the apoptotic process. In summary, the association can enhance the compound's antitumor effects and establish a higher selectivity for tumor cells, possibly caused by significant mitochondrial damage and antioxidant properties.
Asunto(s)
Neoplasias Óseas , Osteosarcoma , Humanos , Animales , Ratones , Vitamina K 3/farmacología , Especies Reactivas de Oxígeno/metabolismo , Apoptosis , Osteosarcoma/tratamiento farmacológico , Osteosarcoma/genética , Combinación de Medicamentos , Línea Celular Tumoral , Neoplasias Óseas/patología , Proliferación CelularRESUMEN
Phenolic phytochemicals are a group of organic compounds with potent antioxidant features but can also act as powerful pro-oxidants. These characteristics are effective in reducing metastatic potential in cancer cells, and this effect has been associated with reactive oxygen species (ROS). Methyl vanillate (MV) and its dimer, methyl divanillate (DMV), are potent antioxidants. In the present study, we investigated the effects of MV and DMV on breast cancer cell lines MCF-7 and MDA-MB-231 and compared the results using the non-tumor cell line HB4a. Our results indicated that the compounds performed a pro-oxidant action, increasing the generation of ROS. DMV decreased the viability cell, showing a higher apoptotic effect and inhibition of proliferation than MV on both cell lines, with significant differences between groups (p < 0.05). Some modulation of NOX4, NOX5, and DUOX were observed, but the results did not correlate with the intracellular production of ROS. The dimer showed more effectivity and pro-oxidant effect than MV, impacting cell line MCF-7 in higher extension than MDA-MB-231. In conclusion, and corroborating with reported works, the dimerization of natural phenolic compounds was associated with improved beneficial biological effects as a potential cytotoxic agent to tumor cells.
Asunto(s)
Neoplasias de la Mama , Antioxidantes/metabolismo , Antioxidantes/farmacología , Apoptosis , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Proliferación Celular , Dimerización , Femenino , Humanos , Células MCF-7 , Especies Reactivas de Oxígeno/metabolismo , Ácido Vanílico/análogos & derivadosRESUMEN
Myrcia bella Cambess (Myrtaceae) is an important and common plant, native to the Brazilian Cerrado, with cytotoxicity, antimicrobial, and antidiabetic properties. Therefore, the effects of crude hydroalcoholic extract (CE) and fractions of ellagitannins (ELT) and flavonoids (FV) from Myrcia bella leaves were evaluated in a UMR-106 murine osteosarcoma cells and MC3T3 (normal cell). Cell viability and migration, production of reactive oxygen species (ROS) and matrix metalloproteinase (MMP) -2 and -9 activities were evaluated. In general, CE (80 µg/mL), ELT (160 µg/mL) and FV (64 µg/mL) reduced cell viability (p < 0.05). FV (64 µg/mL) was more effective in inhibition of cell migration, ROS production, and MMP-2 activity when compared to CE and ELT. Myrcia bella a rich source of phenolic compounds and its fraction of flavonoids have cytotoxic effects on osteosarcoma cells, preserving the viability of normal osteoblasts. Due to its antioxidant capacity, flavonoid may be a new therapeutic strategy for cancer.
Asunto(s)
Myrtaceae , Osteosarcoma , Ratones , Animales , Flavonoides/farmacología , Taninos/farmacología , Especies Reactivas de Oxígeno , Extractos Vegetales/farmacología , Antioxidantes/farmacología , Fenoles/farmacología , Hojas de la Planta , Taninos Hidrolizables/farmacología , Osteosarcoma/tratamiento farmacológicoRESUMEN
The extract of Myracrodruon urundeuva Allemão (aroeira), as a vehicle, associated with calcium hydroxide (CH) paste was evaluated based on cell viability, antimicrobial action, calcium ion release, and pH variation. Calcium hydroxide with propylene glycol was used as control. The pH variation was measured at 3, 24, 72, 168, 140, 360, and 720 h and calcium ion release was measured on days 7, 15, and 30. Cell viability was assessed with NIH/3T3 cells using MTT and crystal violet assays, after 24, 48, and 72 h. Antibacterial activity was determined by the disc diffusion method, while microbial reduction (Enterococcus faecalis) was evaluated using the time-kill test. The CH paste formulated with aroeira showed antibacterial activity against Enterococcus faecalis and further did not interfere with pH, calcium ion release, or cell viability; moreover, the formulation had antimicrobial activity and could serve as a vehicle for CH paste.
Asunto(s)
Anacardiaceae , Hidróxido de Calcio , Animales , Antibacterianos/farmacología , Hidróxido de Calcio/farmacología , Enterococcus faecalis , Concentración de Iones de Hidrógeno , Ratones , Extractos Vegetales/farmacologíaRESUMEN
Abstract Background: Qualea grandiflora (QG) (Vochysiaceae), also known as "pau-ferro", "pau-terra" or "pau-de-tucano", is a very common deciduous tree in the Brazilian Cerrado used in traditional medicine to treat inflammations, ulcers, diarrhea, and infections. There are reports in the scientific literature that demonstrate the medicinal effects of the bark and leaf of the QG. However, studies involving this plant are rather imited. Aim of the study: To perform the phytochemical analysis of the QG hydroalcoholic extract (HAE) of leaves, and to investigate it effects on fibroblast and preosteoblasts. Methods: Phytochemical analysis was done by HPLC-DAD. Murine NIH/3T3 fibroblasts and MC3T3-E1 preosteoblasts cell lines (ATCC) were used for the experiments. Cell viability was assessed by the MTT colorimetric assay and the expression of MMP-14 and HIF-1α by immunofluorescence. Results and conclusion: The following compounds were identified by HPLC-DAD, such as quinic acid, ethyl galate, ellagic acid derivatives as O-methylellagic acid O-galloyl, O-methylellagic acid O-deoxyhexoside, galloyl derivatives, flavonol glycoside as kaempferol-O-deoxyhexoside, quercetin-O-deoxyhexoside, myricetin-O-deoxyhexoside and the pentacyclic triterpene arjunglucoside. Cell viability results demonstrated no cytotoxic effects in the studied concentrations. We found in QG HAE some compounds with therapeutic properties that can increase the expression of MMP-14 and HIF-1α, in fibroblasts and preosteoblasts. These data suggest that QG HAE has an action on these two molecules widely involved in physiological conditions, such as collagen remodeling, bone development and growth and pathological processes as HIF signaling in cancer metastasis.
RESUMEN
Bone development and healing processes involve a complex cascade of biological events requiring well-orchestrated synergism with bone cells, growth factors, and other trophic signaling molecules and cellular structures. Beyond health processes, MMPs play several key roles in the installation of heart and blood vessel related diseases and cancer, ranging from accelerating metastatic cells to ectopic vascular mineralization by smooth muscle cells in complementary manner. The tissue inhibitors of MMPs (TIMPs) have an important role in controlling proteolysis. Paired with the post-transcriptional efficiency of specific miRNAs, they modulate MMP performance. If druggable, these molecules are suggested to be a platform for development of "smart" medications and further clinical trials. Thus, considering the pleiotropic effect of MMPs on mammals, the purpose of this review is to update the role of those multifaceted proteases in mineralized tissues in health, such as bone, and pathophysiological disorders, such as ectopic vascular calcification and cancer.
Asunto(s)
Remodelación Ósea/fisiología , Matriz Extracelular/fisiología , Metaloproteinasas de la Matriz/fisiología , Enfermedades Óseas/metabolismo , Enfermedades Óseas/fisiopatología , Progresión de la Enfermedad , Humanos , Inhibidores de la Metaloproteinasa de la Matriz/uso terapéutico , Neoplasias/metabolismo , Neoplasias/fisiopatología , Osteoblastos/fisiología , Inhibidores Tisulares de Metaloproteinasas/fisiología , Calcificación Vascular/metabolismo , Calcificación Vascular/fisiopatologíaRESUMEN
A espécie vegetal Qualea grandiflora (QG), popularmente conhecida como "pauferro", "pau-terra-da-folha-grande", "pau-terra" ou "pau-de-tucano", muito comum no Cerrado brasileiro, é bem conhecida devido às suas variadas propriedades terapêuticas. Suas indicações incluem ações preventivas no aparecimento de lesões de mucosa gástrica, efeitos analgésicos, antibacterianos, anti-inflamatórios e antifúngicos. Assim, os componentes da QG poderiam ter alguma ação sobre moléculas amplamente envolvidas em processos angiogênicos e de desenvolvimento/reparo, como a Metaloproteinase de matriz 14 (MMP-14) e o Fator Induzido por hipóxia 1α (HIF-1alfa). Dessa maneira, o objetivo deste estudo foi investigar os efeitos do extrato hidroalcoólico das folhas de QG na viabilidade celular e expressão de MMP-14 e HIF-1alpha em culturas de fibroblastos da linhagem NIH/3T3 e pré-osteoblastos da linhagem MC3T3-E1. Para o teste de viabilidade celular e expressão das moléculas, concentrações de 0.1; 1.0 e 10 µg/mL do extrato hidroalcoólico das folhas de QG foram administrados por períodos de 24, 48, 72 e 96h. Após cada período, a viabilidade celular foi avaliada pelo método de redução de MTT e a análise da expressão das moléculas foi feita por meio da técnica de imunofluorescência. Os resultados mostram que o extrato de QG não promove redução da viabilidade celular de fibroblastos e pré-osteoblastos em concentrações até 10 µg/mL, nos períodos iniciais (24 e 48h). Porém, uma redução significativa da viabilidade pode ser verificada nos períodos de 72h e 96h para os fibroblastos e 96h para os pré-osteoblastos, expostos a mais alta concentração do extrato (10 µg/mL). O ensaio de imunofluorescência indica que o extrato, nas concentrações de 0.1; 1.0 e 10 µg/mL foi capaz de aumentar a expressão de MMP-14 e HIF-1alpha, em ambos os tipos celulares. Em conclusão, nossos resultados indicam que o extrato de QG exerce um efeito capaz de aumentar a expressão das duas moléculas em estudo (MMP-14 e HIF-1alpha), tanto para os fibroblastos da linhagem NIH/3T3 como para os pré- osteoblastos da linhagem MC3T3-E1. Assim, os compostos de QG podem apresentar potencial para serem utilizados como agentes terapêuticos moduladores da angiogênese, por meio do aumento da expressão de MMP-14 e HIF-1alpha.(AU)
The vegetable specie Qualea grandiflora (QG), popularly known as "pau-ferro", "pauterra-da-folha-grande", "pau-terra" or "pau-de-tucano", very common in the Brazilian Cerrado, is well known due to its varied therapeutic properties. Its indications include preventive actions in the appearance of lesions of gastric mucosa, analgesic, antibacterial, anti-inflammatory and antifungal effects. Thus, QG components could have some action on molecules widely involved in angiogenic and developmental / repair processes, such as Matrix metalloproteinase 14 (MMP-14) and HypoxiaInducible Factor-1α (HIF-1alpha). Thus, the objective of our study was to investigate the effects of QG hydroalcoholic extract on cell viability and expression of MMP-14 and HIF-1alpha in NIH/3T3 fibroblasts and MC3T3-E1 pre-osteoblasts cell lines. For the cell viability assay and expression of the molecules, concentrations of 0.1; 1.0 and 10 µg / mL of the hydroalcoholic extract of leaves of QG, were administered for periods of 24, 48, 72 and 96h. After each period, the cell viability was evaluated by MTT assay and the expression of the molecules was analyzed using the immunofluorescence technique. The results show that the QG extract does not promote reduction of the cellular viability of fibroblasts and pre-osteoblasts in concentrations up to 10 µg/mL in the initial periods (24 and 48h). However, a significant reduction in viability can be observed in 72h and 96h for fibroblasts and 96h for pre-osteoblasts exposed to the highest extract concentration (10 µg/mL). The immunofluorescence assay indicates that the extract, at concentrations of 0.1; 1.0 and 10 µg/mL was able to increase the expression of MMP-14 and HIF-1alpha in both cell types. In conclusion, our results indicate that the QG extract exerts an effect capable of increasing the expression of the two molecules under study (MMP-14 and HIF-1alpha) both for the NIH/3T3 fibroblasts as well as for the MC3T3-E1 pre-osteoblasts cells. Thus, the QG compounds could have potential to be used as angiogenesis modulating therapeutic agents, by increasing the expression of MMP-14 and HIF-1alpha.(AU)