5 research outputs found
The REST Gene Signature Predicts Drug Sensitivity in Neuroblastoma Cell Lines and Is Significantly Associated with Neuroblastoma Tumor Stage
Neuroblastoma is the most common and deadly solid tumor in children, and there is currently no effective treatment available for neuroblastoma patients. The repressor element-1 silencing transcription (REST) factor has been found to play important roles in the regulation of neural differentiation and tumorigenesis. Recently, a REST signature consisting of downstream targets of REST has been reported to have clinical relevance in both breast cancer and glioblastoma. However it remains unclear how the REST signature works in neuroblastoma. Publicly available datasets were mined and bioinformatic approaches were used to investigate the utility of the REST signature in neuroblastoma with both preclinical and real patient data. The REST signature was found to be associated with drug sensitivity in neuroblastoma cell lines. Further, neuroblastoma patients with enhanced REST activity are significantly associated with higher clinical stages. Loss of heterozygosity on chromosome 11q23, which occurs in a large subset of high-risk neuroblastomas, tends to be correlated with high REST activity, with marginal significance. In conclusion, the REST signature has important implications for targeted therapy, and it is a prognostic factor in neuroblastoma patients
Identification of Ubr1 as an amino acid sensor of steatosis in liver and muscle
Abstract Background Malnutrition is implicated in human metabolic disorders, including hepatic steatosis and myosteatosis. The corresponding nutrient signals and sensors as well as signalling pathways have not yet been well studied. This study aimed to unravel the nutrientāsensing mechanisms in the pathogenesis of steatosis. Methods Plin2, a lipid droplet (LD) proteināinhibiting lipolysis, is associated with steatosis in liver and muscle. Taking advantage of the Gal4āUAS system, we used the Drosophila melanogaster wing imaginal disc as an in vivo model to study the regulation of Plin2 proteostasis and LD homeostasis. Drosophila Schneider 2 (S2) cells were used for western blotting, immunoprecipitation assays, amino acidābinding assays and ubiquitination assays to further investigate the regulatory mechanisms of Plin2 in response to nutrient signals. Mouse AML12 hepatocytes, human JHHā7 and SNUā475 hepatoma cells were used for immunofluorescence, western blotting and immunoprecipitation to demonstrate that the mode of Plin2 regulation is evolutionarily conserved. In addition, we purified proteins from HEK293 cells and reconstituted in vitro cellāfree systems in amino acidābinding assays, pulldown assays and ubiquitination assays to directly demonstrate the molecular mechanism by which Ubr1 senses amino acids to regulate Plin2 proteostasis. Results As a lipolysis inhibitor, Plin2 was significantly elevated in liver (PĀ <Ā 0.05) and muscle (PĀ <Ā 0.05) in patients with steatosis. Consistently, we found that the ubiquitin moiety can be conjugated to any Lys residue in Plin2, ensuring robust clearance of Plin2 by protein degradation. We further demonstrated that the E3 ubiquitin ligase Ubr1 targets Plin2 for degradation in an amino acidādependent manner. Ubr1 uses two canonical substrateābinding pockets, independent of each other, to bind basic and bulky hydrophobic amino acids, respectively. Mechanistically, amino acid binding allosterically activates Ubr1 by alleviating Ubr1's autoāinhibition. In the absence of amino acids, or when the amino acidābinding capacity of Ubr1 is diminished, Ubr1āmediated Plin2 degradation is inactivated, leading to steatosis. Conclusions We identified Ubr1 as an amino acid sensor regulating Plin2 proteostasis, bridging the knowledge gap between steatosis and nutrient sensing. Our work may provide new strategies for the prevention and treatment of steatosis
Protein kinaseĀ C and protein kinaseĀ A are involved in the protection of recombinant human glucagonālike peptideā1 on glomeruli and tubules in diabetic rats
Abstract Aims/Introduction Blockade or reversal the progression of diabetic nephropathy is a clinical challenge. The aim of the present study was to examine whether recombinant human glucagonālike peptideā1 (rhGLPā1) has an effect on alleviating urinary protein and urinary albumin levels in diabetic rats. Materials and Methods Streptozotocināinduced diabetes rats were treated with rhGLPā1 insulin and saline. Using immunostaining, hematoxylināeosin, electron microscopy and periodic acidāSchiff staining to study the pathology of diabetic nephropathy, and we carried out quantitative reverse transcription polymerase chain reaction, western blot and immunohistochemistry to identify the differentially expressed proteins. The mechanism was studied through advanced glycation endāproductsāinduced tubular epithelial cells. Results rhGLPā1 inhibits protein kinaseĀ C (PKC)āĪ², but increases protein kinaseĀ A (PKA), which reduces oxidative stress in glomeruli and in cultured glomerular microvascular endothelial cells. In tubules, rhGLPā1 increased the expression of two key proteins related to reāabsorption ā megalin and cubilin ā which was accompanied by downregulation of PKCāĪ² and upregulation of PKA. On human proximal tubular epithelial cells, rhGLPā1 enhanced the absorption of albumin, and this was blocked by a PKC activator or PKA inhibitor. Conclusions These findings suggest that rhGLPā1 can reverse diabetic nephropathy by protecting both glomeruli and tubules by inhibiting PKC and activating PKA