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Impeded autophagy can impair pancreatic ß cell function by causing apoptosis, of which DAP-related apoptosis-inducing kinase-2 (DRAK2) is a critical regulator. Here, we identified a marked up-regulation of DRAK2 in pancreatic tissue across humans, macaques, and mice with type 2 diabetes (T2D). Further studies in mice showed that conditional knockout (cKO) of DRAK2 in pancreatic ß cells protected ß cell function against high-fat diet feeding along with sustained autophagy and mitochondrial function. Phosphoproteome analysis in isolated mouse primary islets revealed that DRAK2 directly phosphorylated unc-51-like autophagy activating kinase 1 (ULK1) at Ser56, which was subsequently found to induce ULK1 ubiquitylation and suppress autophagy. ULK1-S56A mutation or pharmacological inhibition of DRAK2 preserved mitochondrial function and insulin secretion against lipotoxicity in mouse primary islets, Min6 cells, or INS-1E cells. In conclusion, these findings together indicate an indispensable role of the DRAK2-ULK1 axis in pancreatic ß cells upon metabolic challenge, which offers a potential target to protect ß cell function in T2D.
Assuntos
Proteínas Reguladoras de Apoptose , Proteína Homóloga à Proteína-1 Relacionada à Autofagia , Diabetes Mellitus Tipo 2 , Células Secretoras de Insulina , Hipernutrição , Proteínas Serina-Treonina Quinases , Animais , Humanos , Camundongos , Apoptose , Autofagia , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Células Secretoras de Insulina/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Reguladoras de Apoptose/metabolismoRESUMO
BACKGROUND: WD40 transcription factors are crucial in plant growth and developmental, significantly impacting plant growth regulation. This study investigates the WD40 transcription factor HmWDR68's role in developing the distinctive blue infertile flower colors in Hydrangea macrophylla 'Forever Summer'. METHODS AND RESULTS: The HmWDR68 gene was isolated by PCR, revealing an open reading frame of 1026 base pairs, which encodes 341 amino acids. Characterized by four WD40 motifs, HmWDR68 is a member of the WD40 family. Phylogenetic analysis indicates that HmWDR68 shares high homology with PsWD40 in Camellia sinensis and CsWD40 in Paeonia suffruticosa, both of which are integral in anthocyanin synthesis regulation. Quantitative real-time PCR (qRT-PCR) analysis demonstrated that HmWDR68 expression in the blue infertile flowers of 'Forever Summer' hydrangea was significantly higher compared to other tissues and organs. Additionally, in various hydrangea varieties with differently colored infertile flowers, HmWDR68 expression was markedly elevated in comparison to other hydrangea varieties, correlating with the development of blue infertile flowers. Pearson correlation analysis revealed a significant association between HmWDR68 expression and the concentration of delphinidin 3-O-glucoside, as well as key genes involved in anthocyanin biosynthesis (HmF3H, HmC3'5'H, HmDFR, and HmANS) in the blue infertile flowers of 'Forever Summer' hydrangea (P < 0.01). CONCLUSION: These findings suggest HmWDR68 may specifically regulate blue infertile flower formation in hydrangea by enhancing delphinidin-3-O-glucoside synthesis, modulating expression of HmF3H, HmC3'5'H, HmDFR and HmANS. This study provides insights into HmWDR68's role in hydrangea's blue flowers development, offering a foundation for further research in this field.
Assuntos
Antocianinas , Hydrangea , Antocianinas/genética , Hydrangea/química , Hydrangea/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Filogenia , Pigmentação/genética , Flores/metabolismo , Glucosídeos/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Oxygen and nutrient deprivation are common features of solid tumors. Although abnormal alternative splicing (AS) has been found to be an important driving force in tumor pathogenesis and progression, the regulatory mechanisms of AS that underly the adaptation of cancer cells to harsh microenvironments remain unclear. Here, we found that hypoxia- and nutrient deprivation-induced asparagine endopeptidase (AEP) specifically cleaved DDX3X in a HIF1A-dependent manner. This cleavage yields truncated carboxyl-terminal DDX3X (tDDX3X-C), which translocates and aggregates in the nucleus. Unlike intact DDX3X, nuclear tDDX3X-C complexes with an array of splicing factors and induces AS events of many pre-mRNAs; for example, enhanced exon skipping (ES) in exon 2 of the classic tumor suppressor PRDM2 leads to a frameshift mutation of PRDM2. Intriguingly, the isoform ARRB1-Δexon 13 binds to glycolytic enzymes and regulates glycolysis. By utilizing in vitro assays, glioblastoma organoids, and animal models, we revealed that AEP/tDDX3X-C promoted tumor malignancy via these isoforms. More importantly, high AEP/tDDX3X-C/ARRB1-Δexon 13 in cancerous tissues was tightly associated with poor patient prognosis. Overall, our discovery of the effect of AEP-cleaved DDX3X switching on alternative RNA splicing events identifies a mechanism in which cancer cells adapt to oxygen and nutrient shortages and provides potential diagnostic and/or therapeutic targets.
Assuntos
Processamento Alternativo , Glioblastoma , Animais , Humanos , RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/metabolismo , Glioblastoma/patologia , Oxigênio/metabolismo , Isoformas de Proteínas/metabolismo , Splicing de RNA , Fatores de Processamento de RNA/metabolismo , Microambiente TumoralRESUMO
Rhododendron mole (Blume) G. Don is an attractive ornamental and valuable medicinal plant which widely distributed in the southern regions of China. In order to promote the studies on the genetic diversity of this species, we assembled the complete chloroplast (cp) genome of R. molle by using the genome skimming approach. The results showed that the cp genome of R. molle exhibited a quadripartite cycle with 197,877 bp, comprising of two inverted repeats (IRs) of 43,831 bp separated by a large single copy (LSC) region of 110,189 bp and a quite small single copy (SSC) region of 26 bp. It encodes 146 genes, including 92 protein-coding, 46 tRNA, and eight rRNA genes. The overall GC content of the cp genome was 36.0%. The phylogenetic analysis indicated that R. molle is closely related to R. delavayi. Thus, the cp genome sequence of R. molle provides a rich source of genetic information for studies on Rhododendron taxonomy, phylogeny, and evolution, as well as lays the foundation for further development and utilization of R. molle.
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The current study aimed to investigate the relation of circulating tumor cell (CTC) with clinicopathological features. In addition, its longitudinal change during chemotherapy and its correlation with prognosis in advanced gallbladder carcinoma (GBC) patients were explored. Totally 45 unresectable, locally advanced or metastatic GBC patients who underwent chemotherapy were enrolled in this prospective study. The CTC in 7.5 ml blood was detected at pre-treatment and 3 months post-treatment. CTC was almost detectable in all advanced GBC patients before treatment, whose count was positively correlated with metastatic disease (vs. local advanced disease) (P=0.002), number of organs with metastases (P=0.006), and CA199 level (P=0.002). After treatment, CTC count declined from 4.0 (range: 0.0-83.0) at pre-treatment to 2.0 (range: 0.0-36.0) at post-treatment (P=0.003). Interestingly, pre-treatment CTC count (P=0.270) was of no difference, while post-treatment CTC count was lower (P=0.038) in objective-response patients compared to that in non-objective-response patients; meanwhile, both pre-treatment CTC count (P=0.017) and post-treatment CTC count (P<0.001) were lower in disease-control patients compared with those in non-disease-control patients. Importantly, pre-treatment CTC count ≥2 (versus <2) was only correlated with worse progression-free survival (PFS) (P=0.014) but not overall survival (OS) (P=0.057); while pre-treatment CTC count ≥5 (versus <5), post-treatment CTC count ≥2 (versus <2), post-treatment CTC count ≥5 (versus <5), CTC count up (versus equal/down) were all correlated with poor PFS and OS (all P<0.050). In conclusion, higher CTC count during chemotherapy correlates with worse treatment response, PFS and OS in advanced GBC patients, which implies that CTC measurement may optimize the prognostication and individualized treatment in these patients.
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BACKGROUND: The development of gallbladder disease (GBD) is related to bile acid (BA) metabolism, and the rate of BA circulation increases the risk of biliary cancer. However, it is unclear whether patterns of circulating bile acids (BAs) change in patients with benign GBDs such as gallbladder stones and polyps. Herein, we compared and characterised plasma BA profiles in patients with cholecystolithiasis and non-neoplastic polyps with healthy controls, and explored relationships between plasma BA profiles, demographics, and laboratory test indices. METHODS: A total of 330 subjects (13 healthy controls, 292 cholecystolithiasis and 25 non-neoplastic polyps) were recruited and plasma BA profiles including 14 metabolites from patients with pathologically confirmed cholecystolithiasis and non-neoplastic polyps were compared with controls. BAs were quantitated by liquid chromatography and mass spectrometry, and statistical and regression analyses of demographics and laboratory test indices were performed. RESULTS: Females displayed a higher burden of GBD than males (63.36% cholecystolithiasis, 60% non-neoplastic polyps). Cholecystolithiasis and non-neoplastic polyps were associated with increased plasma total secondary BAs, while levels of primary BAs were lower than in healthy controls. Plasma ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (TUDCA), glycyurdeoxycholic acid (GUDCA), taurochenodeoxycholic acid (TCDCA) and glycochenodeoxycholic acid (GCDCA) were decreased significantly in GBDs, and ursodeoxycholic acid (UDCA) was negatively correlated with white blood cell count and neutrophil percentage. CONCLUSIONS: Secondary BA levels were higher in patients with cholecystolithiasis and non-neoplastic polyps. White blood cell count and percentage of neutrophil in peripheral blood were negatively correlated with UDCA, indicating an anti-inflammation effect of UDCA.
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Cálculos Biliares , Pólipos , Ácidos e Sais Biliares , Feminino , Humanos , MasculinoRESUMO
Breast cancer is one of the most common cancers in women worldwide. In the past decades, many advances have been made in understanding and treating breast cancer. However, due to the highly heterogeneous nature of this disease, a precise characterization of breast cancer on the molecular level is of great importance but not yet readily available. In the present study, we systematically profiled proteomes and N-glycoproteomes of cancerous, paracancerous, and distal noncancerous tissues from patients with breast cancer. The data revealed distinct proteomic and N-glycoproteomic landscapes between different tissues, showing biological insights obtained from the two data sets were complementary. Specifically, the complement and angiogenesis pathways in the paracancerous tissues were activated. Taken together, the changes that occurred in paracancer tissue and N-glycoproteomics are important complements to the conventional proteomic analysis of cancer tissue. Their combination provides more precise and sensitive molecular correlates of breast cancer. Our data and strategy shed light on precisely defining breast cancer, providing valuable information for individual patient diagnosis and treatment. The MS data of this study have been deposited under the accession number IPX0001924000 at iProX.
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Neoplasias da Mama , Proteômica , Neoplasias da Mama/diagnóstico , Feminino , Humanos , ProteomaRESUMO
To investigate whether SENP1 could play a regulating role in the liver fibrosis process, the Sprague-Dawley (SD) rats were used to establish the liver fibrosis rat models by intraperitoneally injecting with 1 ml/kg of 10% CCl4, while the control normal rats were injected with olive oil. Then confirmation experiments to verify the successful establishment of these models were conducted by detecting the cellular and lobular architecture, and liver function indexes using hematoxylin-eosin staining, Masson's trichrome staining and microplate method, respectively. In addition, the expression levels of fibrosis markers including collagen I, collagen III, α-SMA and TGF-ß1 were inspected using quantitative real-time PCR (qRT-PCR), as well as SMAD2. Subsequently, the relative mRNA and protein level of SENP1 was also determined via qRT-PCR and western blot analysis. Next, the HSC-T6 cells of SENP1 knock-down were constructed and used to test the relative protein expression levels of α-SMA and SMAD2 in these cells. The results of hematoxylin-eosin staining, Masson's trichrome staining and microplate method turned out that the rat liver fibrosis models were constructed successfully, which was further confirmed by the increased expression of collagen I, collagen III, α-SMA and TGF-ß1 in mRNA and protein level, as well as SMAD2. Then the expression of SENP1 was overexpressed in the rat liver fibrosis models induced by CCl4 and the TGF-ß1 treatment could increase the protein expression level of collagen I, collagen III and α-SMA. Lastly, the SENP1 knockdown HSC-T6 cells were successfully constructed, while the silence of SENP1 down-regulated the protein expression of α-SMA and SMAD2. In conclusion, this study provided a new regulation mechanism about the liver fibrosis process.
Assuntos
Endopeptidases/metabolismo , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Fígado/metabolismo , Fígado/patologia , Proteína Smad2/metabolismo , Animais , Linhagem Celular , Cisteína Endopeptidases , Regulação para Baixo , Ratos , Ratos Sprague-DawleyRESUMO
Pancreatic cancer (PC) is an aggressive malignancy with one of the worst mortality rates in the world. Multiple factors, including a complex and poorly understood pathophysiology and difficulty in early detection and diagnosis make successful treatment of pancreatic cancer extremely challenging. In this study, we first detected the expressions of eight selected miRNAs which are predicted repress GLUT1 expression by targeting 3'UTR. We found miR-148a had a significantly down regulated expression and miR-148a level has an inverse correlation with the expression of GLUT1 in PC tissues. Subsequently, examined by dual-luciferase assay and western blot, we confirmed that miR-148a suppressed GLUT1 expression by directly targeting 3'UTR of GLUT1 mRNA. Finally, biological study in two pancreatic cancer cell lines indicates that miR-148a function as a tumor suppressor gene through repressing pancreatic cancer cell proliferation, migration and invasion. We first identified miR-148a, which has down regulated expression in pancreatic cancer tissues, plays as a cancer inhibitor through targeting GLUT1. This study sheds light on the roles of miRNAs in the pathogenesis of pancreatic cancer and may be helpful for clinical diagnosis and treatment.