RESUMEN
Glioma is the most common primary malignant tumor of the central nervous system and has a poor prognosis. Therefore, exploring the key molecular targets is a new opportunity for basic research and clinical treatment of glioma. Previous studies found that circRNA-hsa_circ_0073237 was upregulated in gliomas. Our further analyses of the biological function and molecular mechanism of hsa_circ_0073237 showed that hsa_circ_0073237 was also upregulated in glioma cell lines and could combine with miR-345 to inhibit its expression. miR-345 was downregulated in glioma tissues and cells, and targeted to regulate the expression of hepatoma-derived growth factor (HDGF), while HDGF expression was enhanced in glioma. Hsa_circ_0073237 promoted the expression of HDGF in glioma cells by adsorbing miR-345. Hsa_circ_0073237 siRNA, miR-345, and HDGF siRNA effectively inhibited cell viability and invasion and promoted cell apoptosis. When expression of hsa_circ_0073237 and miR-345 was inhibited simultaneously, cell viability, apoptosis, and invasion did not change significantly; however, after transfection with HDGF overexpression vector, the effects of hsa_circ_0073237 siRNA and miR-345 on glioma cell viability, apoptosis, and invasion were obviously reversed. Further construction of glioma xenograft models in nude mice confirmed that the introduction of miR-345 in vivo effectively inhibited tumor growth, significantly reduced tumor diameter and weight, and obviously decreased the expression of HDGF. Therefore, hsa_circ_0073237 can regulate the biological functions of glioma cells through miR-345/HDGF, thereby affecting the progression of tumors, indicating that the hsa_circ_0073237/miR-345/HDGF pathway may be a key target for the treatment of glioma.
Asunto(s)
Glioma , MicroARNs , Animales , Movimiento Celular , Proliferación Celular/genética , Regulación Neoplásica de la Expresión Génica , Glioma/genética , Péptidos y Proteínas de Señalización Intercelular , Ratones , Ratones Desnudos , MicroARNs/genética , ARN CircularRESUMEN
Glioblastoma multiforme (GBM) is the most frequent, lethal and aggressive tumour of the central nervous system (CNS) in adults. Multidrug resistance (MDR) results in undesirable prognosis during GBM chemotherapy. In this study, we determined that Radicol (RAD), a novel trinorguaiane-type sesquiterpene originally isolated from the root of Dictamnus radicis Cortex, exhibited potently cytotoxic effect on temozolomide (TMZ)-resistant GBM cell lines in a dose-dependent manner. Radicol-induced apoptosis was confirmed with Hoechst 33342/propidium iodide and terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end-labelling (TUNEL) staining. Studies investigating the mechanism revealed that RAD triggered an attenuation of protein disulphide isomerase (PDI) and induced the unmitigated unfolded protein response (UPR) and lethal endoplasmic reticulum (ER) stress. Simultaneously, we further demonstrated that RAD suppressed the activation of Akt/mTOR/p70S6K phosphorylation by up-regulating the induction of glycogen synthase kinase-3ß (GSK-3ß). These results established a link between RAD-induced ER stress and inhibition of the Akt/mTOR/p70S6K pathway, and the attenuation of PDI and activation of GSK-3ß might be the synergistic target of antineoplastic effects during RAD-induced apoptosis. These findings suggested that RAD, possessing multiple cytotoxicity targets, low molecular weight and high lipid solubility, could be a promising agent for the treatment of malignant gliomas. Copyright © 2017 John Wiley & Sons, Ltd.
Asunto(s)
Antineoplásicos/farmacología , Dacarbazina/análogos & derivados , Resistencia a Antineoplásicos , Glioma/patología , Sesquiterpenos/farmacología , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Dacarbazina/farmacología , Dictamnus/química , Estrés del Retículo Endoplásmico/efectos de los fármacos , Glioblastoma/tratamiento farmacológico , Glucógeno Sintasa Quinasa 3/metabolismo , Humanos , Etiquetado Corte-Fin in Situ , Fosforilación/efectos de los fármacos , Fitoterapia , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Sesquiterpenos/química , Sesquiterpenos/clasificación , Serina-Treonina Quinasas TOR , TemozolomidaRESUMEN
Plant-derived natural products have been the highly significant sources of novel antitumor agents. The cassane-type diterpenes of genus Caesalpinia have been reported to bear antiproliferative activities toward different types of cancer cells. In this study, we evaluated the antineoplasmic activities of 16 natural origin cassane-type diterpenes isolated from the CHCl3 extract of the seeds of C. minax in pituitary adenomas cells and identified caesalpin G (CAG) showed the strongest cytotoxicity. Moreover, we further investigated the structure-activity relationship and molecular mechanism of these derivatives systematically. The results confirmed the unsaturated lactone-type ring, hydroxyl at C-7, and alkenyl at C-11 or C-14 functionality as critical for anticancer activity in this family of natural products. In addition, the mechanism experiments also demonstrated unfolded protein response and ER stress and Wnt/ß-catenin pathway were involved in the CAG-induced apoptosis.
Asunto(s)
Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Caesalpinia/química , Diterpenos/aislamiento & purificación , Diterpenos/farmacología , Medicamentos Herbarios Chinos/farmacología , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Diterpenos/química , Ensayos de Selección de Medicamentos Antitumorales , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/aislamiento & purificación , Humanos , Estructura Molecular , Neoplasias Hipofisarias/tratamiento farmacológico , Semillas/química , Relación Estructura-Actividad , Proteínas Wnt/efectos de los fármacos , beta Catenina/efectos de los fármacosRESUMEN
Glioblastoma (GBM) is the most frequent and aggressive tumour in the central nervous system. Many studies have demonstrated that upregulation of the NF-κB onco-pathway is accompanied by the acquisition of Temozolomide (TMZ) resistance in GBM cells. Here, we show that RGFP109, a selective histone deacetylase (HDAC1 and HDAC3) inhibitor, overcomes TMZ resistance and downregulates the expression of NF-κB-regulated pro-survival genes in a TMZ-resistant (TR) GBM cell line. RGFP109 did not alter the phosphorylation levels of NF-κB/p65 or inhibitory κBα (IκBα). Immunofluorescence microscopy showed that RGFP109 does not block the nuclear translocation of NF-κB/p65. However, co-immunoprecipitation assays revealed that RGFP109 induces the hyperacetylation of NF-κB/p65 and histones, and blocks interactions between NF-κB/p65 and its coactivators, p300 and p300/CBP-associated factor (PCAF). These results indicate that RGFP109-mediated post-translational nuclear acetylation may be involved in the regulation of NF-κB. Electrophoretic mobility shift assays revealed that RGFP109 reduces NF-κB/p65 binding to κB-DNA and decreased the transcriptional level of κB-mediated genes, suggesting that RGFP109-induced hyperacetylation leads to attenuated transcription of the κB gene. In addition, RGFP109 elevates the expression of inhibitor of growth 4 (ING4), which is typically downregulated in GBM cells. Importantly, we found that RGFP109 enhances ING4 recognition and binding to NF-κB/p65, which may be positively correlated with reduced interactions between NF-κB/p65 and p300/PCAF, thereby effecting transcription of the κB gene. Finally, we show that knockdown of ING4 with plasmids containing pcDNA3.1-ING4 shRNA abolished the effect of RGFP109. Therefore, ING4 may act as a corepressor and facilitate RGFP109-triggered suppression of the NF-κB pathway. Taken together, our data show that RGFP109, an HDAC inhibitor, in combination with TMZ may be a therapeutic candidate for patients with temozolomide-resistant GBM.
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Antineoplásicos Alquilantes/farmacología , Benzamidas/farmacología , Dacarbazina/análogos & derivados , Resistencia a Antineoplásicos/efectos de los fármacos , Inhibidores de Histona Desacetilasas/farmacología , FN-kappa B/metabolismo , Acetilación , Transporte Activo de Núcleo Celular , Apoptosis , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Núcleo Celular/metabolismo , Neoplasias del Sistema Nervioso Central , Dacarbazina/farmacología , Sinergismo Farmacológico , Glioblastoma , Proteínas de Homeodominio/metabolismo , Humanos , FN-kappa B/genética , Transducción de Señal , Temozolomida , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Transcripción Genética , Activación Transcripcional , Proteínas Supresoras de Tumor/metabolismoRESUMEN
Arbuscular mycorrhizal (AM) symbiosis is known to stimulate plant drought tolerance. However, the molecular basis for the direct involvement of AM fungi (AMF) in plant water relations has not been established. Two full-length aquaporin genes, namely GintAQPF1 and GintAQPF2, were cloned by rapid amplification of cDNA 5'- and 3'-ends from an AMF, Glomus intraradices. Aquaporin localization, activities and water permeability were examined by heterologous expression in yeast. Gene expression during symbiosis was also analyzed by quantitative real-time polymerase chain reaction. GintAQPF1 was localized to the plasma membrane of yeast, whereas GintAQPF2 was localized to both plasma and intracellular membranes. Transformed yeast cells exhibited a significant decrease in cell volume on hyperosmotic shock and faster protoplast bursting on hypo-osmotic shock. Polyethylene glycol (PEG) stimulated, but glycerol inhibited, the aquaporin activities. Furthermore, the expression of the two genes in arbuscule-enriched cortical cells and extraradical mycelia of maize roots was also enhanced significantly under drought stress. GintAQPF1 and GintAQPF2 are the first two functional aquaporin genes from AMF reported to date. Our data strongly support potential water transport via AMF to host plants, which leads to a better understanding of the important role of AMF in plant drought tolerance.
Asunto(s)
Acuaporinas/genética , Genes Fúngicos/genética , Glomeromycota/genética , Micorrizas/genética , Calcio/metabolismo , Clonación Molecular , Recuento de Colonia Microbiana , Biología Computacional , Sequías , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación de la Expresión Génica de las Plantas , Glomeromycota/crecimiento & desarrollo , Datos de Secuencia Molecular , Micelio/genética , Micorrizas/crecimiento & desarrollo , Presión Osmótica , Filogenia , Pichia/crecimiento & desarrollo , Pichia/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/microbiología , Transporte de Proteínas/genética , Protones , Protoplastos/metabolismo , Estrés Fisiológico/genética , Transformación Genética , Agua/metabolismo , Zea mays/genética , Zea mays/microbiologíaRESUMEN
Medicinal plants have been used world-wide for thousands of years and are widely recognized as having high healing but minor toxic side effects. The scarcity and increasing demand for medicinal plants and their products have promoted the development of artificial cultivation of medicinal plants. Currently, one of the prominent issues in medicinal cultivation systems is the unstable quality of the products. Arbuscular mycorrhiza (AM) affects secondary metabolism and the production of active ingredients of medicinal plants and thus influence the quality of herbal medicines. In this review, we have assembled, analyzed, and summarized the effects of AM symbioses on secondary metabolites of medicinal plants. We conclude that symbiosis of AM is conducive to favorable characteristics of medicinal plants, by improving the production and accumulation of important active ingredients of medicinal plants such as terpenes, phenols, and alkaloids, optimizing the composition of different active ingredients in medicinal plants and ultimately improving the quality of herbal materials. We are convinced that the AM symbiosis will benefit the cultivation of medicinal plants and improve the total yield and quality of herbal materials. Through this review, we hope to draw attention to the status and prospects of, and arouse more interest in, the research field of medicinal plants and mycorrhiza.
Asunto(s)
Hongos/fisiología , Micorrizas/fisiología , Extractos Vegetales/metabolismo , Plantas Medicinales/metabolismo , Plantas Medicinales/microbiología , Simbiosis , Extractos Vegetales/análisis , Plantas Medicinales/químicaRESUMEN
Soil respiration and extracellular enzyme activity are important components of the material cycle of mountain ecosystems and play key roles in maintaining ecosystem functions. To explore the coupling relationship between soil functions and environmental factors, the soil functional indicators, environmental factors, and effects of altitude on the soil function of 36 soil samples from 12 altitudes of the Meili Mountain were analyzed. The results showed that there were significant differences in soil respirations and enzyme activities among altitudes of Meili Mountain, and high-altitude areas had higher soil functions. Soil functions increased with altitudinal difference. PCA analysis showed that the first three axes explained 56.7%, 17.4%, and 8.7% of the variance in soil functional elevation change, respectively, indicating that the functional changes related to carbon and phosphorus were higher than those related to nitrogen. There were significant correlations between environmental factors and soil functional indicators; soil function indicators had stronger correlations with soil physicochemical properties than with climatic factors. Altitude mainly affected soil function indirectly by affecting soil physicochemical properties and climatic factors. These results have great scientific significance for improving the understanding of the material cycle and ecological function of the Meili Mountain ecosystem and provide an important reference for in-depth study of the altitude distribution pattern and evolution characteristics of the soil function of the mountain ecosystem.
RESUMEN
Arbuscular mycorrhizal fungi (AMF) play key roles in enhancing plant tolerance to heavy metals, and iron (Fe) compounds can reduce the bioavailability of arsenic (As) in soil, thereby alleviating As toxicity. However, there have been limited studies of the synergistic antioxidant mechanisms of AMF (Funneliformis mosseae) and Fe compounds in the alleviation of As toxicity on leaves of maize (Zea mays L.) with low and moderate As contamination. In this study, a pot experiment was conducted with different concentrations of As (0, 25, 50 mgêkg-1) and Fe (0, 50 mgêkg-1) and AMF treatments. Results showed that under low and moderate As concentrations (As25 and As50), the co-inoculation of AMF and Fe compound significantly increased the biomass of maize stems and roots, phosphorus (P) concentration, and P-to-As uptake ratio. Moreover, the co-inoculation of AMF and Fe compound addition significantly reduced the As concentration in stem and root, malondialdehyde (MDA) content in leaf, and soluble protein and non-protein thiol (NPT) contents in leaf of maize under As25 and As50 treatments. In addition, co-inoculation with AMF and Fe compound addition significantly increased the activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in the leaves of maize under As25 treatment. Correlation analysis showed that stem biomass and leaf MDA content were very significantly negatively correlated with stem As content, respectively. In conclusion, the results indicated that the co-inoculation of AMF and Fe compound addition can inhibit As uptake and promote P uptake by maize under low and moderate As contamination, thereby mitigating the lipid peroxidation on maize leaves and reducing As toxicity by enhancing the activities of antioxidant enzymes under low As contamination. These findings provide a theoretical basis for the application of AMF and Fe compounds in the restoration of cropland soil contaminated with low and moderate As.
RESUMEN
The crystallization of alkane melts on carbon nanotubes (CNT) and the surface of graphene nanosheets (GNS) is investigated using molecular dynamics (MD) simulations. The crystallization process of the alkane melts is analyzed in terms of the bond-orientational order parameter, atomic radial distribution for the CNT/alkane, atomic longitudinal distribution for the GNS/alkane, and diffusion properties. The dimensional effects of the different carbon-based nanostructures on the crystallization of alkane melts are shown. It is found that one-dimensional CNT has a stronger ability to induce the crystallization of the polymer than that of two-dimensional GNS, which provides a support at molecular level for the experimental observation [Li et al., J. Am. Chem. Soc., 2006, 128, 1692 and Xu et al., Macromolecules, 2010, 43, 5000]. From the MD simulations, we also find that the crystallization of alkane molecules has been completed with the highly cooperative processes of adsorption and orientation.
RESUMEN
Arbuscular mycorrhizal (AM) symbiosis facilitates plant mineral nutrient acquisition and plays key roles in plant adaptation to environmental stresses. The application of AM fungi is a component of sustainable agriculture and ecological restoration. We introduced the current status of AM fungi collections, production of commercial inocula and AM fungi related patents, summarized the research advances in inoculum production, inoculation techniques, and factors influencing the success of inoculation practice in the field, based on case studies of mycorrhizal technology in agriculture, horticulture, and ecological restoration. Finally, we proposed some basic scientific questions and technical bottleneck that deserve futher studies, to promote the development and application of mycorrhizal technologies.
Asunto(s)
Micorrizas , Agricultura , Raíces de Plantas , Plantas , SimbiosisRESUMEN
Effects of an arbuscular mycorrhizal (AM) fungus (Glomus caledonium) on accumulation and metabolism of atrazine in maize grown in soil contaminated with different concentrations of atrazine were investigated in a series of pot experiments. Roots of mycorrhizal plants accumulated more atrazine than non-mycorrhizal roots. In contrast, atrazine accumulation in shoot decreased in mycorrhizal compared with non-mycorrhizal plants. No atrazine derivatives were detected in the soil, either with or without mycorrhizal colonization. However, atrazine metabolites, deethylatrazine (DEA) and deisopropylatrazine (DIA), were detected in plant roots and the AM colonization enhanced the metabolism. After plant harvest atrazine concentrations decreased markedly in the soils compared to the initial concentrations. The decreases were the most in rhizosphere soil and then near-rhizosphere soil and the least in bulk soil. Mycorrhizal treatment enhanced atrazine dissipation in the near-rhizosphere and bulk soils irrespective of atrazine application rates.
Asunto(s)
Atrazina/farmacocinética , Herbicidas/análisis , Micorrizas/metabolismo , Suelo/análisis , Zea mays/metabolismo , Atrazina/análogos & derivados , Atrazina/análisis , Atrazina/metabolismo , Biomasa , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Contaminantes del Suelo/análisis , Triazinas/análisisRESUMEN
The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment. Non-mycorrhizal and zero-P addition controls were included. Plant biomass and concentrations and uptake of As, P, and other nutrients, AM colonization, root lengths, and hyphal length densities were determined. The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium. Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments, but shoot and root biomass of AM plants was depressed by P application. AM fungal inoculation decreased shoot As concentrations when no P was added, and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition, respectively. Shoot and root uptake of P, Mn, Cu, and Zn increased, but shoot Fe uptake decreased by 44.6%, with inoculation, when P was added. P addition reduced shoot P, Fe, Mn, Cu, and Zn uptake of AM plants, but increased root Fe and Mn uptake of the nonmycorrhizal ones. AM colonization therefore appeared to enhance plant tolerance to As in low P soil, and have some potential for the phytostabilization of As-contaminated soil, however, P application may introduce additional environmental risk by increasing soil As mobility.
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Arsénico/metabolismo , Micorrizas/metabolismo , Fósforo/farmacología , Zea mays/microbiología , Biodegradación Ambiental/efectos de los fármacos , Concentración de Iones de Hidrógeno , Micorrizas/efectos de los fármacos , Micorrizas/crecimiento & desarrollo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Microbiología del Suelo , Contaminantes del Suelo/metabolismo , Zea mays/efectos de los fármacos , Zea mays/metabolismoRESUMEN
A strategy to suppress the expression of the DNA repair enzyme O6methylguanineDNA methyltransferase (MGMT) by inhibition of Wnt/ßcatenin signaling may be useful as a novel treatment for pituitary adenoma. Previous studies have reported that Tanshinone IIA (TSA), a major quinone compound isolated from Salvia miltiorrhiza, had antitumor effects. However, whether TSA has antitumor effects against pituitary adenoma and whether the mechanisms are associated with the Wnt/ßcatenin/MGMT pathway remains to be clarified. In the present study, TSA treatment caused apoptosis in AtT20 cells in a concentrationdependent manner, as demonstrated by cell viability reduction, phophatidylserine externalization detected by Annexin V staining and mitochondrial membrane potential disruption detected by JC1 staining, which were associated with activation of caspase3 and DNA fragmentation detected by TUNEL in AtT20 cells. Tcell factor (TCF)lymphoidenhancing factor (LEF) reporter activity was determined by dual luciferase reporter assay and the interaction between ßcatenin and TCF4 were detected using a coimmunoprecipitation kit. The results indicated TSA treatment increased ßcatenin phosphorylation, inhibited ßcatenin nuclear translocation, reduced ßcatenin/TCF4 complex formation and TCFLEF luciferase reporter activity, and subsequently reduced the expression of cyclin D1 and MGMT. Notably, overexpression of MGMT in ßcatenin knock down AtT20 cells abrogated the TSAmediated effects in AtT20 cells. In conclusion, TSA induced apoptosis via inhibition of Wnt/ßcatenindependent MGMT expression, which may provide novel insights into the understanding of the mechanism of the antitumor effects of Salvia miltiorrhiza.
Asunto(s)
Abietanos/farmacología , Antineoplásicos Fitogénicos/farmacología , Corticotrofos/efectos de los fármacos , Metilasas de Modificación del ADN/genética , Enzimas Reparadoras del ADN/genética , Regulación Neoplásica de la Expresión Génica , Salvia miltiorrhiza/química , Proteínas Supresoras de Tumor/genética , beta Catenina/genética , Abietanos/aislamiento & purificación , Animales , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Corticotrofos/metabolismo , Corticotrofos/patología , Fragmentación del ADN , Metilasas de Modificación del ADN/antagonistas & inhibidores , Metilasas de Modificación del ADN/metabolismo , Enzimas Reparadoras del ADN/antagonistas & inhibidores , Enzimas Reparadoras del ADN/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Factor de Unión 1 al Potenciador Linfoide/genética , Factor de Unión 1 al Potenciador Linfoide/metabolismo , Ratones , Extractos Vegetales/química , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Factor de Transcripción 4/genética , Factor de Transcripción 4/metabolismo , Proteínas Supresoras de Tumor/antagonistas & inhibidores , Proteínas Supresoras de Tumor/metabolismo , Vía de Señalización Wnt/efectos de los fármacos , beta Catenina/antagonistas & inhibidores , beta Catenina/metabolismoRESUMEN
Triboelectric nanogenerators (TENGs) have been invented recently for meeting the power requirements of small electronics and potentially solving the worldwide energy crisis. Here, we developed a vertical contact-separation mode TENG based on a novel multi-dielectric-layered (MDL) structure, which was comprised of parylene C, polyimide and SiO2 films. By using the corona discharge approach, the surface charge density was enhanced to as high as 283 µC m-2, and especially the open-circuit voltage could be increased by a factor of 55 compared with the original value. Furthermore, the theoretical models were built to reveal the output characteristics and store the electrostatic energy of the TENG. The influences of the structural parameters and operation conditions including the effective dielectric thickness, dielectric constant, gap distance and air breakdown voltage were investigated systematically. It was found that the output performances such as the peak voltage and power density are approximately proportional to the thickness of the MDL film, but they would be restricted by the air breakdown voltage. These unique structures and models could be used to deepen the understanding of the fundamental mechanism of TENGs, and serve as an important guide for designing high performance TENGs.
RESUMEN
Carbon transport in arbuscular mycorrhizal (AM) symbiosis is of fundamental importance. However, the role of glycerol transport in AM symbiosis has not yet been resolved. Glycerol transport across the cell membrane is mediated by aquaglyceroporins (AQGPs), whereas our previous study revealed that it was disfavoured by GintAQPF2, an AQGP from AM fungi (AMF). Here, we analysed the function of two amino acid residues in the aromatic/arginine (ar/R) constriction known as the major selectivity filter in AQGPs. Replacement of phenylalanine-94 (Phe-94) by alanine (Ala) enlarged the diameter of the ar/R constriction and resulted in an increased intracellular glycerol accumulation and thus survival rate of yeast cells at high glycerol levels, while individual or joint replacement of Phe-94 and Ala-234 by tryptophan and glycine induced a closed state of GintAQPF2, suggesting that the potential double gates (Phe94-Phe243 and arginine-249) of the ar/R constriction also likely determined solute permeability. To figure out whether GintAQPF2 functions were relevant to the establishment of AM symbiosis, genomic analyses of four representative fungi with different lifestyles were performed. We found that glycerol facilitators existed in the facultative fungi (the ectomycorrhizal fungus Laccaria bicolor and hemibiotrophic pathogen Magnaporthe oryzae), but not in the obligatory fungi (the AMF Rhizophagus irregularis and necrotrophic pathogen Fusarium verticillioides), revealing a conserved pattern of glycerol transport in symbionts and pathogens. Our results suggested that glycerol blocks due to the special structural features of the ar/R constriction in the only AMF AQGP could potentially play a role in the establishment of AM symbiosis.
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Acuagliceroporinas/genética , Acuagliceroporinas/metabolismo , Glicerol/metabolismo , Micorrizas/enzimología , Micorrizas/metabolismo , Mutación Puntual , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Acuagliceroporinas/química , Secuencia Conservada , Análisis Mutacional de ADN , Modelos Moleculares , Micorrizas/genética , Micorrizas/fisiología , Conformación Proteica , Alineación de SecuenciaRESUMEN
Temozolomide (TMZ) is the cornerstone of therapy for glioblastoma multiforme (GBM). However, its efficacy is limited due to the development of multidrug resistance (MDR). In this study, we first identified the occurrence of ER stress-tolerance (ERST) in glioma cells and confirmed that ERST was positively correlated with TMZ resistance. We further showed that the seesaw-effect of HDAC6-p97/VCP (increased HDAC6 and decreased p97/VCP) in glioma cells was crucial to ERST-associated TMZ resistance. Moreover, the combination treatment of Tubastatin A (TUB, a selective inhibitor of HDAC6) and TMZ synergistically overcame ERST, reduced cell viability and induced apoptosis in TMZ-resistant glioma cells. TUB and TMZ triggered pro-apoptotic signals of the unfolded protein response (UPR) and ER stress and reversed the ratio between HDAC6 and p97/VCP, which potentially attenuated the activation of heat shock proteins and mediated the reversal of ERST. The combination treatment also triggered the dissociation of Dynein-HDAC6 and attenuation of the Dynein-Dynactin motor complex. In addition, this treatment induced HDAC6-p97/VCP-mediated ubiquitination-autophagy turnover, which was involved in the degradation and clearance of ubiquitinated misfolded proteins. This effect could be partially reversed by HDAC6 KO and/or p97/VCP overexpression. Therefore, we proposed that glioma cells optimized the clearance of ubiquitinated misfolded proteins via the reinforcement of HDAC6-facilitated autophagy and attenuation of the p97/VCP-mediated ubiquitin-proteasome system (UPS). In conclusion, our findings showed that the balance of HDAC6-p97/VCP was crucial to ERST-associated TMZ resistance and that HDAC6 inhibition might be a synergistic target and strategy along with TMZ for the improvement of clinical glioma treatment.
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Dacarbazina/análogos & derivados , Inhibidores de Histona Desacetilasas/uso terapéutico , Histona Desacetilasas/uso terapéutico , Ácidos Hidroxámicos/uso terapéutico , Indoles/uso terapéutico , Autofagia , Dacarbazina/administración & dosificación , Dacarbazina/uso terapéutico , Histona Desacetilasa 6 , Inhibidores de Histona Desacetilasas/farmacología , Histona Desacetilasas/farmacología , Humanos , Ácidos Hidroxámicos/farmacología , Indoles/farmacología , Temozolomida , Transfección , UbiquitinaciónRESUMEN
A greenhouse pot experiment was carried out to investigate the effect of the arbuscular mycorrhizal fungus Glomus etunicatum on the uptake of atrazine (ATR) and cadmium (Cd) from soil by maize (Zea mays L.). Mycorrhizal colonization led to an increase in the accumulation of Cd and ATR in maize roots but a decrease in the shoots. Atrazine alleviated the adverse effects of Cd on maize growth, and this was more pronounced in the inoculated plants. An increase in Cd accumulation by maize roots was observed when ATR was also present. After harvest, the residual ATR concentration in the soil decreased markedly. With mycorrhizal inoculation the amount of residual ATR decreased more in the bulk soil but less in the rhizosphere soil compared to the noninoculated controls. Cadmium application significantly decreased the ATR residual concentrations in both the rhizosphere and bulk soils irrespective of inoculation treatment.
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Atrazina/metabolismo , Cadmio/metabolismo , Hongos/metabolismo , Herbicidas/metabolismo , Suelo/análisis , Zea mays/microbiología , Atrazina/análisis , Cadmio/análisis , Hongos/crecimiento & desarrollo , Herbicidas/análisis , Raíces de Plantas/química , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Brotes de la Planta/química , Brotes de la Planta/metabolismo , Brotes de la Planta/microbiología , Zea mays/metabolismoRESUMEN
A glasshouse pot experiment was conducted to investigate the effect of the fungicide chlorothalonil on the growth of upland rice, in the absence or presence of the arbuscular mycorrhizal fungus (AMF) Glomus mosseae (NM and GM treatments). The plants were grown with three concentrations of chlorothalonil (0, 50 and 100 mg kg(-1) soil). Mycorrhizal colonization decreased significantly with increasing chlorothalonil concentrations. Plant biomass decreases were smaller in GM plants than in non-mycorrhizal (NM) plants. Mycorrhizal dependency was the highest with 50 mg kg(-1) chlorothalonil. Chlorothalonil affected physiological processes in upland rice irrespective of inoculation. Chlorothalonil at 50 and 100 mg kg(-1) increased ascorbate peroxidase (APX) activity and soluble protein concentrations in shoots and roots of NM upland rice. However, values of APX, catalase (CAT) and peroxidase (POD) were reduced more in GM plants than in NM plants. These results showed that chlorothalonil induced oxidative stress in upland rice and it is needed to evaluate the side effects of chlorothalonil on rice and AMF.
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Fungicidas Industriales/toxicidad , Micorrizas/fisiología , Nitrilos/toxicidad , Oryza/efectos de los fármacos , Oryza/crecimiento & desarrollo , Oryza/metabolismo , Oryza/microbiología , Proteínas de Plantas/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/microbiologíaRESUMEN
A glasshouse pot experiment was conducted to investigate effects of the arbuscular mycorrhizal fungus Glomus mosseae on the growth of Vicia faba and toxicity induced by heavy metals (HMs) (Cu, Zn, Pb and Cd) in a field soil contaminated by a mixture of these metals. There was also uninoculation treatment (NM) simultaneously. Mycorrhizal (GM) plants hav e significantlyincreased growth and tolerance to toxicity induced by heavy metals compared with NM plants. P uptake was significantly increased in GM plants. Mycorrhizal symbiosis reduced the transportation of HMs from root to shoot by immobilizing HMs in the mycorrhizal, shown by increasing the ratios of HMs from root to shoot. Oxidative stress, which can induce DNA damage, is an important mechanism of heavy metal toxicity. GM treatment decreased oxidative stress by intricating antioxidative systems such as peroxidases and non-enzymic systems including soluble protein. The DNA damage induced by heavy metals was detected using comet assay, which showed DNA damage in the plants was decreased by the GM treatment.
Asunto(s)
Metales Pesados/toxicidad , Micorrizas/fisiología , Microbiología del Suelo , Vicia faba , Ensayo Cometa , Daño del ADN , Estrés Oxidativo , Peroxidasas/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Contaminantes del Suelo/metabolismo , Contaminantes del Suelo/toxicidad , Simbiosis , Vicia faba/efectos de los fármacos , Vicia faba/crecimiento & desarrollo , Vicia faba/metabolismo , Vicia faba/microbiologíaRESUMEN
OBJECTIVE: To study the effects of Arbuscular Mycorrhizae on cultivated Atractylodes lancea. METHOD: Pot experiment of A. lancea, with (code as AM) or without (code as CK) Glomus mosseae (GM) was conducted 5 times respectively, then the biomass, essential oil, and soil nutrition, soil organism, soil microbial were detected after A. lancea were harvested. RESULT: (1) Mycorrhizal dependency of A. lancea was 245%, and height of individuals, numbers of leaves, leaf area, biomass of A. lancea were all higher in AM than in CK (P < 0.05). (2) GC-MS analysis with cluster analysis and principal components analysis showed that there were no differences in essential oil of A. lancea between AM and CK. (T3) Total N, available N, available P and available K in AM soil were all lower than in CK soil. (4) GC-MS analysis showed organic matters changed differenly in AM soil and CK soil, components 5,6 in AM soil were higher than that in CK soil, but component 9, 10, 11 were lower in AM soil than that in CK soil. (5) Biolog detect showed AWCD of AM soil microbe were higher than that of CK soil throughout the incubation, and AWCD of the former was 0.66, and the later was 0.46 after 192 h incubation. and t-test showed, Shannon seven indices and McIntosh'seven indices were same both at 72 h and 192 h, and diversity indices of Shannon and McIntosh were also same at 72 h, but AM soil microbe were higher than CK soil microbe at 168 h (P < 0.05). CONCLUSION: AM could promote nutrition uptake, improve the function diversity and activity of microbe in rhizosphere of A. lancea, influence the composition of the organic matter, that lead the growth of A. lancea, but not to the quality.