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In this study, we assessed the impact of varied water deficit irrigation frequencies (T1: 2.5 L/4 days; T2: 5 L/8 days; CK: 5 L/4 days) on Zitian Seedless grapes from veraison to post-ripening. Notably, total soluble solids increased during on-tree storage compared to at maturity, while total anthocyanin content decreased, particularly in CK (60.16%), T1 (62.35%), and less in T2 (50.54%). Glucose and fructose levels increased significantly in T1 and T2, more so in T2, but slightly declined in CK. Tartaric acid content increased by 41.42% in T2. Moreover, compared to regular irrigation, water deficit treatments enhanced phenolic metabolites and volatile compounds, including chlorogenic acid, various flavonoids, viniferin, hexanal, 2-nonenal, 2-hexen-1-ol, (E)-, 3-hydroxy-dodecanoic acid, and 1-hexanol, etc. Overall, the T2 treatment outperformed T1 and CK in maintaining grape quality. This study reveals that combining on-tree storage with water deficit irrigation not only improves grape quality but also water efficiency.
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Riego Agrícola , Frutas , Vitis , Agua , Vitis/química , Vitis/crecimiento & desarrollo , Vitis/metabolismo , Frutas/química , Frutas/metabolismo , Frutas/crecimiento & desarrollo , Agua/metabolismo , Agua/análisis , Almacenamiento de Alimentos , Antocianinas/análisis , Antocianinas/metabolismo , Fenoles/metabolismo , Fenoles/análisis , Árboles/crecimiento & desarrollo , Árboles/metabolismo , Árboles/química , Flavonoides/análisis , Flavonoides/metabolismoRESUMEN
Herein, the in situ N-alkylation between tripyridine molecules and C2H5OH was employed to obtain four iodometallates (Mn+ = Pb2+, Cu+ and Ag+). Among them, the alkylation degree of the tripyridine molecules is different, and the in situ-generated tripyridine derivatives showed different configurations. This is not only related to the structure of the inorganic anions but also related to the tripyridine molecule itself. The two moieties were actually templates for each other. The Pb2+ compound 1 was found to effectively catalyze the degradation of Rhodamine B (RhB), with its degradation efficiency reaching 94% in 70 min. In contrast, the degradation efficiency for the Cu+ and Ag+ compounds 2-4 did not exceed 50%. The free radical trapping test indicated that with the Pb2+ compound as the photocatalyst, both superoxide ion radicals (ËO2-) and holes (h+) played a key role. Alternatively, with compound 3 or 4 as the photocatalyst, only ËO2- played a role. The differences should be related to the band gap of the materials, where the Pb2+ compound has a slightly wider band gap, leading to a higher exciton separation efficiency. Due to the involvement of more free radicals, the usual intermediates did not appear in the catalytic process of 1, while the colored species appeared in the catalytic process of 3 and 4. The cyclic test verified that compound 1 still had high catalytic activity after recycling five times. Also, we investigated the catalytic capacity of 1 for the degradation of 5-times the quantity of RhB. The degradation rate reached 99% in 5.5 hours, indicating that 1 has a good photocatalytic performance for degrading a high concentration of RhB. However, given that the amount of catalyst and dye used in each study was different, it was difficult to compare the results. Thus, herein, we introduced for the first time the method of calculating the TOF value to better evaluate the photodegradation performance of each hybrid material.
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Anthocyanins have indispensable functions in plant resistance, human health, and fruit coloring, which arouse people's favorite. It has been reported that anthocyanins are widely found in fruits, and can be affected by numerous factors. In this review, we systematically summarize anthocyanin functions, classifications, distributions, biosynthesis, decoration, transportation, transcriptional regulation, DNA methylation, and post-translational regulation in fruits.
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Antocianinas , Frutas , Humanos , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Procesamiento Proteico-Postraduccional , Epigénesis Genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
BES1, as the most important transcription factor responsible for brassinolide (BR) signaling, has been confirmed to play a significant role in regulating plant growth and the improvement of stress resistance. The transcriptional regulatory mechanism of BES1 has been well elucidated in several plants, such as Arabidopsis thaliana (A. thaliana), Triticum aestivum L. (T. aestivum), and Oryza sativa L. (O. sativa). Nevertheless, the genome-wide analysis of the BES1 family in Vitis vinifera L. (V. vinifera). has not been comprehensively carried out. Thus, we have conducted a detailed analysis and identification of the BES1 transcription factors family in V. vinifera; a total of eight VvBES1 genes was predicted, and the phylogenetic relationships, gene structures, and Cis-acting element in their promoters were also analyzed. BES1 genes have been divided into three groups (I, II and III) based on phylogenetic relationship analysis, and most of VvBES1 genes were in group III. Also, we found that VvBES1 genes was located at seven of the total nineteen chromosomes, whereas VvBES1-2 (Vitvi04g01234) and VvBES1-5 (Vitvi18g00924) had a collinearity relationship, and their three copies are well preserved. In addition, the intron-exon model of VvBES1 genes were mostly conserved, and there existed several Cis-acting elements related to stress resistance responsive and phytohormones responsive in BES1s genes promoter. Moreover, the BES1 expressions were different in different V. vinifera organs, and BES1 expressions were different in different V. vinifera varieties under saline-alkali stress and heat stress, the expression of VvBES1 also changed with the prolongation of saline-alkali stress treatment time. The above findings could not only lay a primary foundation for the further validation of VvBES1 function, but could also provide a reference for molecular breeding in V. vinifera.
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Proteínas de Arabidopsis , Arabidopsis , Vitis , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vitis/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Expresión Génica , Regulación de la Expresión Génica de las Plantas , Familia de Multigenes , Proteínas de Unión al ADN/genética , Proteínas de Arabidopsis/genéticaRESUMEN
Atherosclerosis (AS), the formation of plaque lesions in the walls of arteries, causes many mortalities and morbidities worldwide. Currently, achieving site-specific delivery and controlled release at plaques is difficult. Herein, we implemented the strategy of constructing a bionic multifunctional nanoplatform (BM-NP) for targeting and improving plaques. BM-NPs were prepared based on probucol-loaded mesoporous polydopamine (MPDA) carriers and were coated with platelet membranes to impart bionic properties. In vitro experiments confirmed that BM-NPs, which respond to near-infrared (NIR) for drug release, remove reactive oxygen species (ROS), thereby reducing the level of oxidized low-density lipoprotein (ox-LDL) and ultimately helping to inhibit macrophage foaming. In vivo experiments proved that BM-NPs actively accumulated in plaques in the mouse right carotid artery (RCA) ligation model. During treatment, BM-NPs with NIR laser irradiation more effectively reduced the area of plaque deposition and slowed the thickening of the arterial wall intima. More importantly, BM-NPs showed the advantage of inhibiting the increase in triglyceride (TG) content in the body, and good biocompatibility. Hence, our research results indicate that intelligent BM-NPs can be used as a potential nanotherapy to precisely and synergistically improve AS.
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Aterosclerosis , Nanopartículas , Placa Aterosclerótica , Animales , Aterosclerosis/tratamiento farmacológico , Dopamina/uso terapéutico , Liberación de Fármacos , Ratones , Nanopartículas/uso terapéutico , Placa Aterosclerótica/tratamiento farmacológico , Especies Reactivas de Oxígeno/uso terapéuticoRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is a chronic metabolic liver disease closely related to obesity, which has become a global health problem. However, current pharmacological therapies for NAFLD are limited by potential side effects, low effectiveness and poor aqueous solubility. Herein, we designed functionalized drug-albumin nanocomposites (BAM15@BSA NPs), which were prepared by self-assembly of the anti-obesity small-molecule drug (BAM15) and bovine serum albumin (BSA), for treatment of NAFLD. The proposed BAM15@BSA NPs not only improve aqueous solubility and half-life of BAM15 but also exhibit hepatic-targeted capacity and an increased therapeutic efficacy. In vitro experiments revealed that BAM15@BSA NPs possessed excellent biocompatibility, and improved resistance to adipogenesis and reduced lipid accumulation in human hepatocellular carcinoma cells. In vivo, BAM15@BSA NPs showed liver targeting ability and powerful anti-obesity effects without altering body temperature or affecting food intake, and could effectively alleviate hepatic steatosis and improve therapeutic efficacy for NAFLD treatment. The above findings demonstrated that BAM15@BSA NPs potentially served as a safe and effective drug for NAFLD treatment.
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Fármacos Antiobesidad , Nanocompuestos , Enfermedad del Hígado Graso no Alcohólico , Fármacos Antiobesidad/farmacología , Humanos , Hígado , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Obesidad/tratamiento farmacológico , Albúmina Sérica Bovina/metabolismoRESUMEN
Developing intelligent responsive platforms to carry out high-performance therapy is of great interest for the treatment of tumors and their metastases. However, effective drug loading, activity maintenance, off-target leakage, and response to collaborative therapy remain great challenges. Herein, a targeted intelligent responsive mesoporous polydopamine (MPDA) nanosystem was reported for use in gene-mediated photochemotherapy for synergistic tumor treatment. First, the MPDA was surface modified to maintain a positive charge near the surface and to impart active targeting. Then, gambogic acid (GA) was encapsulated in the MPDA, solidified by phase change materials (PCMs), and finally loaded with siRNA by electrostatic interactions to obtain the smart nanodelivery system (PPMD@GA/si). In vitro and in vivo experiments showed that it not only effectively avoids siRNA inactivation and accidental release of GA, but also possesses potential for targeted accumulation to tumor tissue and mild-temperature photothermal therapy and chemotherapy via near infrared (NIR) radiation. Additionally, the release of siRNA could also effectively inhibit tumor invasion and metastasis to realize multimodal synergistic therapy. Overall, our studies provide a promising idea for synergistic tumor and metastasis treatment based on vector construction.
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Nanopartículas , Neoplasias , Humanos , Indoles , Nanopartículas/uso terapéutico , Neoplasias/tratamiento farmacológico , Polímeros/farmacología , ARN Interferente Pequeño/farmacologíaRESUMEN
Diabetic wounds have an extremely complex microenvironment of hyperglycemia, hypoxia and high reactive oxygen species (ROS). Therefore, the regulation and management of this microenvironment may provide a new and improved treatment method for chronic diabetic wound healing. Herein, a glucose/ROS cascade-responsive nanozyme (CHA@GOx) was developed for diabetic wound treatment based on Ce-driven coassembly by a special dual ligand (alendronic acid and 2-methylimidazole) and glucose oxidase (GOx). It possesses superoxide dismutase and catalase mimic activities, which effectively remove excess ROS. In particular, it can catalyze excessive hydrogen peroxide generated by the glucose oxidation reaction to produce oxygen, regulate the oxygen balance of the wound, and reduce the toxic side effects of GOx, thus achieving the purpose of synergistically repairing diabetic wounds. In vitro experiments show that CHA@GOx assists mouse fibroblast migration and promotes human umbilical vein endothelial cell tube formation. In vivo, it can induce angiogenesis, collagen deposition, and re-epithelialization during wound healing in diabetic mice. Taken together, this study indicates that the coassembly of multifunctional nanozymes has implications in diabetic wound healing.
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MOFs with adequate free nitrogen sites have potential applications in dye adsorption and formic acid dehydrogenation. Here, we successfully synthesized a novel 3-D MOF 1 ([(CH3)2NH2][Cd(L)DMA]·0.5DMA·1.5H2O) with a special two-fold interpenetrating framework through a simple solvothermal reaction between CdCl2·1.5H2O and a nitrogen-rich triangular tricarboxylate-based linker (H3L, 4,4',4''-s-triazine-2,4,6-tribenzoic acid). After removing the guest molecules of dimethylacetamide (DMA) and H2O, including the coordinated DMA from 1 by vacuum activation at 423 K, a compound named 1' with a formula of [(CH3)2NH2][Cd(L)] and a similar interpenetrating framework structure was obtained. In comparison with compound 1, the total void volume of 1' is nearly doubled, and thus may provide higher potential for the adsorption of other guest molecules. Notably, the pyridine N atoms located in the middle of the triangular tricarboxylate-based linker are not involved in the coordination with Cd2+, and are all uniformly dispersed throughout the whole framework of the 3-D MOFs. Due to its unique structural features, the 3-D MOF 1' could effectively adsorb the cationic dye MB+ for recycling purposes. The rapid adsorption rate (0.7 × 10-2 g mg-1 min-1) and the relatively high capacity (900 mg g-1) for MB+ demonstrate the potential of 1' in dye adsorption. In addition, 1' may also be used as an effective support to immobilize PdAu NPs via the double-solvent method. The resultant catalyst Pd0.8Au0.2/1' exhibits decent catalytic activity for the dehydrogenation of formic acid with a TOF value of 1854 h-1 at 333 K. The existence of a large void volume and accessible pyridine N atoms provide a suitable environment for achieving a high dispersion of PdAu NPs, thereby leading to the formation of a catalytically active and stable supported noble-metal NP catalyst for H2 generation from formic acid decomposition.
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In this study, two water deficit treatments in the same amount of water but with different frequencies (T1: 2.5 L per 4 d and T2: 5 L per 8 d) were performed on Reliance grapevines from veraison until harvest to explore their effects on grape berries quality under root restriction. Results showed that glucose, fructose and sucrose contents were increased, while malic acid, tartaric acid and citric acid contents were decreased under two treatments. Meanwhile, water deficits also promoted the accumulation of phenylalanine and proline. For phenols, anthocyanins, resveratrol and flavonols contents in the water deficit groups were significantly higher than those in the control group. In addition, two water deficit treatments increased the characteristic aromas contents, especially the esters contents. Overall, T2 treatment had a better effect than T1 treatment. This study provided an idea for improving water use efficiency and grape quality.
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Vitis , Antocianinas , Flavonoles , Frutas , AguaRESUMEN
Root restriction (RR) has been reported to enhance grape berry quality in diverse aspects of grape life. In this study, RR-induced increases in the main primary metabolites in the grape berry and the expression of their related genes were studied at different developmental stages. Mainly the transcriptomic and metabolomic level were analyzed using 'Summer Black' grape berry as a material. The main results were as follows: A total of 11 transcripts involved in the primary metabolic pathways were significantly changed by the RR treatment. Metabolites such as sugars, organic acids, amino acids, starch, pectin, and cellulose were qualitatively and quantitatively analyzed along with their metabolic pathways. Sucrose synthase (VIT_07s0005g00750, VIT_11s0016g00470) and sucrose phosphate synthase (VIT_18s0089g00410) were inferred to play critical roles in the accumulation of starch, sucrose, glucose, and fructose, which was induced by the RR treatment. RR treatment also promoted the malic acid and tartaric acid accumulation in the young berry. In addition, the grape berries after the RR treatment tended to have lower pectin and cellulose content.
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Vitis , Celulosa/metabolismo , Frutas , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Pectinas/metabolismo , Almidón/análisis , Transcriptoma/genética , Vitis/metabolismoRESUMEN
As the incidence of diabetes increases, its complication, diabetic foot ulcers, has become the main type of clinically chronic refractory wounds. Due to the hyperglycemic microenvironment of the diabetic wound, which leads to vascular defects and bacterial growth, the therapeutic effect of wound dressings lacking strategic design is relatively limited. In this study, we designed an injectable, "self-healing", and glucose-responsive multifunctional metal-organic drug-loaded hydrogel (DG@Gel) for diabetic wound healing. The functionalized hydrogel was prepared by phase-transfer-mediated metallo-nanodrugs, which were made by co-assembling zinc ions, organic ligands, and a small-molecule drug, deferoxamine mesylate (DFO), and the programmed loading of glucose oxidase (GOX). When injected into a diabetic wound, the GOX in DG@Gel changed the hyperglycemic wound microenvironment by decomposing excess glucose into hydrogen peroxide and glucuronic acid, which decreased the pH of the wound site. The low pH promoted the release of zinc ions and DFO, which exhibited synergistic antibacterial and angiogenesis activity for diabetic wound repair. In vitro experiments revealed the antibacterial activity and the cell proliferation, migration, and tube formation ability of DG@Gel. Moreover, in vivo experiments showed that DG@Gel can induce re-epithelialization, collagen deposition, and angiogenesis during wound healing in diabetic mice with good biocompatibility and biodegradability. The results suggest that this hydrogel is a promising innovative dressing for the treatment of diabetic wounds. STATEMENT OF SIGNIFICANCE: Diabetic ulcers, as one of the main types of chronic refractory wounds, are not treated effectively in the clinic due to a lack of strategic approach. In this study, we designed a glucose-responsive multifunctional metal-organic drug-loaded hydrogel (DG@Gel), which can change the hyperglycemic wound microenvironment by decomposing excess glucose into hydrogen peroxide and glucuronic acid. This in turn promoted the release of zinc ions and deferoxamine mesylate (DFO) in the hydrogel, which exhibited synergistic antibacterial and angiogenic activity for diabetic wound repair. Furthermore, the DG@Gel exhibited good biocompatibility and biodegradability in vivo. In general, this innovative strategy design may have great application potential in the treatment of various chronic wounds.
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Diabetes Mellitus Experimental , Pie Diabético , Animales , Diabetes Mellitus Experimental/tratamiento farmacológico , Glucosa/farmacología , Hidrogeles/química , Ratones , Cicatrización de HeridasRESUMEN
Ginkgo seeds are a traditional food in China valued for their nutritional and health benefits. However, little is known about the anti-aging and health-promoting effects of ginkgo seed products. Here, we showed that ginkgo seed powder extract (GSP-E) is abundant in alkaloids and flavonoids, and can extend the lifespan of Caenorhabditis elegans. GSP-E improved most physiological indicators related to aging of C. elegans, including locomotor activity, reproductive capacity, and resistance to oxidation and heat. Moreover, GSP-E reduced the accumulation of lipofuscin and reactive oxygen species (ROS) in C. elegans. Further studies demonstrated that GSP-E improved longevity and stress resistance by mediating lipid metabolism and autophagy, as well as by regulating gene expression (e.g., FASN-1, POD-2, GPX-7, FAT-5). GSP-E has an anti-amyloid effect and delayed amyloid-induced paralysis of C. elegans. These findings could support the utilization of ginkgo seed as a potential dietary supplement for the health food industry, and provide a novel health-promoting resource against aging and aging-related diseases.
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Envejecimiento/efectos de los fármacos , Antioxidantes/farmacología , Suplementos Dietéticos , Ginkgo biloba , Extractos Vegetales/farmacología , Animales , Antioxidantes/administración & dosificación , Caenorhabditis elegans/efectos de los fármacos , Alimentos Funcionales , Fitoterapia , Extractos Vegetales/administración & dosificación , SemillasRESUMEN
Upregulation of transmembrane protein 97 (TMEM97) has been associated with progression and poor outcome in multiple human cancers, including breast cancer. Recent studies suggest that TMEM97 may be involved in the activation of the Wnt/ß-catenin pathway. However, the molecular mechanism of TMEM97 action on Wnt/ß-catenin signaling is completely unclear. In the current study, TMEM97 was identified as an LRP6-interacting protein. TMEM97 could interact with LRP6 intracellular domain and enhance LRP6-mediated Wnt signaling in a CK1δ/ε-dependent manner. The binding of TMEM97 to LRP6 facilitated the recruitment of CK1δ/ε to LRP6 complex, resulting in LRP6 phosphorylation at Ser 1490 and the stabilization of ß-catenin. In breast cancer cells, knockout of TMEM97 attenuated the Wnt/ß-catenin signaling cascade via regulating LRP6 phosphorylation, leading to a decrease in the expression of Wnt target genes AXIN2, LEF1, and survivin. TMEM97 deficiency also suppressed cell viability, proliferation, colony formation, migration, invasion, and stemness properties in breast cancer cells. Importantly, TMEM97 knockout suppressed tumor growth through downregulating the Wnt/ß-catenin signaling pathway in a breast cancer xenograft model. Taken together, our results revealed that TMEM97 is a positive modulator of canonical Wnt signaling. TMEM97-mediated Wnt signaling is implicated in the tumorigenesis of breast cancer, and its targeted inhibition may be a promising therapeutic strategy for breast cancer.
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Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Proteína-6 Relacionada a Receptor de Lipoproteína de Baja Densidad/metabolismo , Proteínas de la Membrana/metabolismo , Oncogenes , Vía de Señalización Wnt , Animales , Quinasa de la Caseína I/metabolismo , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Supervivencia Celular , Regulación hacia Abajo , Femenino , Genes Reporteros , Células HEK293 , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Fosforilación , Fosfoserina/metabolismo , Unión Proteica , Ensayo de Tumor de Célula Madre , Ensayos Antitumor por Modelo de Xenoinjerto , beta Catenina/metabolismoRESUMEN
Resveratrol is a natural polyphenol compound produced in response to biotic and abiotic stresses in grape berries. However, changes in resveratrol caused by bud sport in grapes are scarcely reported. In this study, trans-resveratrol and cis-resveratrol were identified and quantified in the grape berries of 'Summer Black' and its bud sport 'Nantaihutezao' from the veraison to ripening stages using ultra performance liquid chromatography-high resolution tandem mass spectrometry (UPLC-HRMS). We found that bud sport accumulates the trans-resveratrol earlier and increases the contents of cis-resveratrol in the earlier stages but decreases its contents in the later stages. Simultaneously, we used RNA-Seq to identify 51 transcripts involved in the stilbene pathways. In particular, we further identified 124 and 19 transcripts that negatively correlated with the contents of trans-resveratrol and cis-resveratrol, respectively, and four transcripts encoding F3'5'H that positively correlated with the contents of trans-resveratrol by weighted gene co-expression network analysis (WGCNA). These transcripts may play important roles in relation to the synergistic regulation of metabolisms of resveratrol. The results of this study can provide a theoretical basis for the genetic improvement of grapes.
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The compositions and contents of metabolites in the pulp tissue play critical roles in the fruit quality for table grape. In this study, the effects of root restriction (RR) on the primary and secondary metabolites of pulp tissue at five developmental stages were studied at the metabolomics level, using "Red Alexandria" grape berry (Vitis vinifera L.) as materials. The main results were as follows: 283 metabolites were annotated by using ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS); 28 and 16 primary metabolites contents were increased and decreased, and 11 and 19 secondary metabolites contents were increased and decreased, respectively, along the berry development; RR significantly decreased 12 metabolites (four amino acids and derivatives, three organic acids, four flavonoids and one other compound) contents, and improved 40 metabolites (22 amino acids and derivatives, six nucleotides, four carbohydrates, four cofactors, three cinnamic acids and one other compound) accumulation at the different developmental stages. Altogether, our study would be helpful to increase our understanding of grape berry's responses to RR stress.
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The precise operation of the hypoxic tumor microenvironment presents a promising way to improve treatment efficacy, in particular in tumor synergistic phototherapy. This work reports an innovative approach to build adenosine triphosphate-modified hollow ceria nanozymes (ATP-HCNPs@Ce6) that manipulate tumor hypoxia to effectively achieve drug delivery. Hollow ceria nanoparticles (HCNPs) exhibit a controllable hollow structure through varying nitric acid concentrations in the nanocomposites. Specifically, ATP modification makes HCNPs exceptionally biocompatible and stable and acts as a regulator of HCNP enzymatic activity. In the stage of drug loading, newly prepared ATP-HCNPs@Ce6 serves as an in situ oxygen-generating agent because of its ability to simulate catalase. Therefore, ATP-HCNPs@Ce6 has adjustable enzymatic properties that act like a "switch" to selectively supply oxygen in response to high levels of hydrogen peroxide expression and the slightly acidic lysosomal environment of the tumor to enhance lysosome-targeted photodynamic therapy. Moreover, the obvious anticancer effects of ATP-HCNPs@Ce6 are demonstrated in vitro and in vivo. Overall, a simple and rapid self-assembly strategy to form and modify multifunctional HCNPs is reported, which may further propel their application in the field of precision tumor treatment.
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Materiales Biomiméticos/química , Catalasa/metabolismo , Cerio/química , Lisosomas/metabolismo , Terapia Molecular Dirigida/métodos , Nanopartículas/química , Fototerapia/métodos , Adenosina Trifosfato/química , Línea Celular Tumoral , Humanos , Peróxido de Hidrógeno/metabolismoRESUMEN
BACKGROUND: Targeting ubiquitin-dependent proteolysis is one of the strategies in cancer therapy. CRLCDT2 and CRLDDB2 are two key E3 ubiquitin ligases involved in DNA replication and DNA damage repair. But CDT2 and DDB2 are opposite prognostic factors in kinds of cancers, and the underlining mechanism needs to be elucidated. METHODS: Small interfering RNAs were used to determine the function of target genes. Co-immunoprecipitation (Co-IP) was performed to detect the interaction between DDB2 and CDT2. Immunofluorescence assays and fluorescence activating cell sorting (FACS) were used to measure the change of DNA content. In vivo ubiquitination assay was carried out to clarify the ubiquitination of CDT2 mediated by DDB2. Cell synchronization was performed to arrest cells at G1/S and S phase. The mechanism involved in DDB2-mediated CDT2 degradation was investigated by constructing plasmids with mutant variants and measured by Western blot. Immunohistochemistry was performed to determine the relationship between DDB2 and CDT2. Paired two-side Student's t-test was used to measure the significance of the difference between control group and experimental group. RESULTS: Knockdown of DDB2 stabilized CDT2, while over-expression of DDB2 enhanced ubiquitination of CDT2, and subsequentially degradation of CDT2. Although both DDB2 and CDT2 contain PIP (PCNA-interacting protein) box, PIP box is dispensable for DDB2-mediated CDT2 degradation. Knockdown of PCNA had negligible effects on the stability of CDT2, but promoted accumulation of CDT1, p21 and SET8. Silencing of DDB2 arrested cell cycle in G1 phase, destabilized CDT1 and reduced the chromatin loading of MCMs, thereby blocked the formation of polyploidy induced by ablation of CDT2. In breast cancer and ovarian teratoma tissues, high level of DDB2 was along with lower level of CDT2. CONCLUSIONS: We found that CRL4DDB2 is the novel E3 ubiquitin ligases of CDT2, and DDB2 regulates DNA replication through indirectly regulates CDT1 protein stability by degrading CDT2 and promotes the assembly of pre-replication complex. Our results broaden the horizon for understanding the opposite function of CDT2 and DDB2 in tumorigenesis, and may provide clues for drug discovery in cancer therapy.
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MAIN CONCLUSION: 4-coumarate-CoA ligase (VIT_02s0109g00250) and copper amine oxidase (VIT_17s0000g09100) played essential roles in contributing to the total soluble solid and total anthocyanin variations induced by bud sport in grape berries. Taste and color, which are important organoleptic qualities of grape berry, undergo rapid and substantial changes during development and ripening. In this study, we used two cultivars 'Summer Black' and its bud sport 'Nantaihutezao' to explore and identify differentially expressed genes associated with total soluble solid and anthocyanin during developmental stages using RNA-Seq. Overall, substantial differences in expression were observed across berry development between the two cultivars. 5388 genes were detected by weighted gene co-expression network analysis (WGCNA) associated with the total soluble solid (TSS) and anthocyanin contents variations. Several of these genes were significantly enriched in the phenylalanine metabolism pathway; two hub genes 4-coumarate-CoA ligase (VIT_02s0109g00250) and copper amine oxidase (VIT_17s0000g09100) played the most essential roles in relating to the total soluble solid and total anthocyanin variations induced by bud sport through Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment and co-expression network analysis. These findings provide insights into the molecular mechanism responsible for the bud sport phenotype.
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Frutas , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Transcriptoma , Vitis , Frutas/enzimología , Frutas/genética , Frutas/crecimiento & desarrollo , Perfilación de la Expresión Génica , Genes de Plantas/genética , Transcriptoma/genética , Vitis/enzimología , Vitis/genética , Vitis/crecimiento & desarrolloRESUMEN
In the present study, the effects of root restriction (RR) on the main phenolic metabolites and the related gene expression at different developmental stages were studied at the transcriptomic and metabolomic levels in "Summer Black" grape berries (Vitis vinifera × Vitis labrusca). The results were as follows: seven phenolic acid compounds, three stilbene compounds, nine flavonol compounds, 10 anthocyanin compounds, and 24 proanthocyanidin compounds were identified by ultra-performance liquid chromatography-high-resolution mass spectrometry. RR treatment significantly promoted the biosynthesis of phenolic acid, trans-resveratrol, flavonol, and anthocyanin and also affected the proanthocyanidin content, which was elevated in the early developmental stages and then reduced in the late developmental stages. The functional genes for phenylalanine ammonia-lyase, trans-cinnamate 4-monooxygenase, 4-coumarate-CoA ligase, shikimate O-hydroxycinnamoyl transferase, chalcone synthase, chalcone isomerase, stilbene synthase, flavonoid 3',5'-hydroxylase, anthocyanidin 3-O-glucosyltransferase, and the transcription factors MYBA1, MYBA2, MYBA3, and MYBA22 were inferred to play critical roles in the changes regulated by RR treatment.