关键词： 脑膜瘤   微小RNA-494   激活转录因子3   预后  

摘要： 目的分析微小RNA-494(miR-494)、激活转录因子3信使RNA(ATF3 mRNA)在脑膜瘤组织中的表达及其意义。方法选取2013年10月—2018年1月于首都医科大学附属北京友谊医院神经外科脑膜瘤切除患者79例为脑膜瘤组,另取因脑外伤切除脑组织患者79例为对照组。采用实时荧光定量PCR(qRT-PCR)法检测2组miR-494、ATF3 mRNA表达水平;分析脑膜瘤组织miR-494、ATF3 mRNA表达水平与脑膜瘤患者临床病理特征及预后的关系;Pearson法分析脑膜瘤组织miR-494表达水平与ATF3 mRNA的相关性;COX回归分析脑膜瘤患者预后的影响因素。结果脑膜瘤组miR-494、ATF3 mRNA表达水平均显著高于对照组(t/P=7.038/0.000、8.271/0.000)。脑膜瘤组织miR-494、ATF3 mRNA低表达亚组有淋巴结转移、WHO分级Ⅲ级、TNM分期Ⅲ~Ⅳ期患者比例低于高表达亚组(miR-494:χ^(2)/P=5.346/0.021、6.627/0.010、12.381/0.000;ATF3 mRNA:χ^(2)/P=12.868/0.000、14.799/0.000、12.166/0.000);脑膜瘤组织miR-494表达水平与ATF3 mRNA水平呈正相关(r/P=0.578/0.000);脑膜瘤患者miR-494、ATF3 mRNA低表达亚组术后36个月累积生存率分别明显高于miR-494、ATF3 mRNA高表达亚组(χ^(2)/P=21.798/0.019、13.015/0.000)。多因素分析结果显示,发生淋巴结转移、TNM分期增加及miR-494、ATF3 mRNA表达水平升高是影响脑膜瘤患者预后的独立危险因素[OR(95%CI)=2.572(1.582~4.182)、2.463(1.554~3.904)、2.257(1.478~3.447)、2.385(1.547~3.678)]。结论脑膜瘤组织中miR-494、ATF3 mRNA表达水平显著升高,两者可能相互作用共同影响脑膜瘤发生发展,检测miR-494、ATF3 mRNA水平有利于判定患者预后情况。

关键词： Citrus alkaline extracts   Pulmonary fibrosis   Endoplasmic reticulum stress   ATF3   PINK1   DEGRADATION   PINK1   LUNG  

摘要： Background: Idiopathic pulmonary fibrosis is a chronic, progressive, fibrotic disease. Although the pathogenesis remains unclear, the effect of endoplasmic reticulum (ER) stress in type II alveolar epithelial cells (AEC IIs) is increasingly thought to be a critical mechanism. Purpose: We investigated the effects of citrus alkaline extracts (CAE) on AEC IIs and elucidated the underlying mechanism for their possible use in ameliorating pulmonary fibrosis (PF). Methods: A bleomycin-induced mouse model of PF, and an in vitro tunicamycin (TM)-induced ER stress model in A549 cells were successfully established. Accumulation of collagen in lung tissues in vivo was assessed using histological analysis and western blotting. The expression levels of the ER-stress marker BiP and other related proteins were assessed by western blotting and immunofluorescence staining. Mitochondrial membrane potential was assessed to evaluate mitochondrial homeostasis. Results: CAE mitigated collagen deposition to ameliorate PF in vivo. CAE suppressed the bleomycin or TM-induced increases in ER-stress biomarker, BiP, and PERK pathway proteins, resulting in a decrease in ER stress in mouse lung tissues and A549 cells, respectively. Additionally, CAE treatment suppressed the bleomycin or TM-induced increase in the ER-stress downstream proteins, activating ATF3 and increased the levels of PINK1 in AEC IIs, both in vivo and in vitro. The reduced mitochondrial homeostasis induced by TM was restored by CAE-treatment in A549 cells. Furthermore, conditioned media from TM-treated A549 cells increased collagen deposition in MRC5 cells mainly via TGF-beta 1. The increased collagen deposition was not seen using conditioned media from CAE-treated A549 cells. Conclusion: These results provide novel insights into the potential mechanism of CAE in inhibiting ER stress in AEC IIs, and suggests that it has great potential to ameliorate PF via the ATF3/PINK1 pathway.

关键词： FOXI1   High-throughput data   Chromatin immunoprecipitation   ATF3   miR-590   TRANSCRIPTION FACTOR-3   METASTASIS   PROMOTES   P53   LINC00941   GENE  

摘要： FOXI1 plays a key role in the development of gastric cancer. However, the whole genome FOXI1 binding sites and its target genes are unclear. In the present study, we used ChIP-seq and RNA-seq technologies to identify the target gene of FOXI1. Firstly, ChIP-seq data showed that, 4476 unique peaks in the genome region were captured. Most of these binding peaks are located in introns or intergenic regions. We annotated all the peaks to the nearest gene and identified 404 genes as FOXI1 binding genes. KEGG and GO analysis showed that FOXI1 binding gene to be correlated with the cellular process, cell part, cell, binding, single-organism process. Further, we performed FOXI1-overexpressed RNA-seq experiment. We comprehensively analyzed the ChIP-seq and RNA-seq data and take the intersection of two databases, several genes were identified. ATF3 was selected from the intersection since ATF3 was the most enriched mRNA after FOXI1 overexpressed. ChIP-qPCR and luciferase report gene were used to validate that ATF3 was target gene of FOXI1. Intriguely, ATF3 protein was significantly downregulated after FOXI1 overex -pressed. We found FOXI1 can also bind to the promoter of miR-590 and active it which directly target ATF3. The binding site between FOXI1 and miR-590 was verified by ChIP-qPCR and luciferase report gene, and the target relationship between miR-590 and ATF3 was confirmed by dual-luciferase reporter gene. In conclusion, our data identified the genome binding sites of FOXI1, and provide evidence that FOXI1 inhibits gastric cancer cell proliferation by activating miR-590/ATF3 axis. (c) 2021 Published by Elsevier Ltd.

关键词： Arsenic   ATF3   Transformation   Inflammation   Lung cancer   ACTIVATING TRANSCRIPTION FACTOR-3   LUNG-CANCER   DOWN-REGULATION   AKT ACTIVATION   CONTRIBUTES   EXPRESSION   AUTOPHAGY   DISEASE   STRESS   IL6  

摘要： Arsenic is a naturally occurring metalloid strongly associated with the incidence of lung cancer. Understanding the mechanisms of arsenic-induced carcinogenesis favors the development of effective interventions to reduce the incidence and mortality of lung cancer. In this study, we investigated the role of activating transcription factor 3 (ATF3) in arsenic-induced transformation of human bronchial epithelial cells. ATF3 was upregulated during chronic exposure to 0.25 mu M arsenic, and loss of ATF3 promoted arsenic-induced transformation. Moreover, arsenic-transformed ATF3 knockout (ATF3 KO-AsT) cells exhibited more aggressive characteristics, including acceleration in proliferation, resistance to chemotherapy and increase in migratory capacity. RNA-seq revealed that pathways involved in inflammation, cell cycle, EMT and oncogenesis were affected due to ATF3 deficiency during chronic arsenic exposure. Further experiments confirmed the overproduction of IL-6, IL-8 and TNF alpha as well as enhanced phosphorylation of AKT and STAT3 in ATF3 KO-AsT cells. Our results demonstrate that ATF3 upregulated by chronic low-dose arsenic exposure represses cell transformation and acquisition of malignant characteristics through inhibiting the production of proinflammatory cytokines and activation of downstream proteins AKT and STAT3, providing a new strategy for the prevention of carcinogen-induced lung cancer.

关键词： Ovarian   ATF3   Immune cell infiltration   Bioinformatical analysis   CANCER METASTASIS   TRANSCRIPTION   CYTOSKELETON   INFLAMMATION  

摘要： ATF3 is an essential transcription activator in regulating cancer-related genetic expression. To identify the role of ATF3 in ovarian tumor, we investigated the correlation between ATF3 expression and the clinicopathological properties using multiple database. The cBioPortal and GEPIA database displayed the clinical information of ovarian patients harboring or without harboring ATF3 mutation. Furthermore, we assessed the relationship between survival and ATF3 expression level using Kaplan-Meier plotter, which reveals that the ovarian patients with higher expression of ATF3 suffered the worse overall survival and progression-free survival. The differentially expressed genes were analyzed using gene ontology, protein-protein interaction network, and gene set enrichment analysis to identify the hub gene and critical pathways, significantly affecting the tumorigenesis of ovarian tumor. Finally, we assessed the correlation between ATF3 and immune cell infiltration using Tumor Immunoassay Resource (TIMER) database. The results demonstrated that higher expression has a positive correlation with macrophage infiltration, expression for M1- and M2-type macrophages. Our study suggests that ATF3 can regulate the cell cycle and heme-related oxidative phosphorylation process, and it may be a critical factor to regulate the macrophage cell to be infiltrated into ovarian cancer. ATF3 can be used as a biomarker for diagnosis and therapy of ovarian tumor.

关键词： Atherosclerosis   ATF3   Plaque stability   Inflammatory reaction   Coronary heart disease   GENE-EXPRESSION   INFLAMMATION   ATF3   BINDING   INTERLEUKIN-1-BETA   PATHOPHYSIOLOGY   MECHANISMS   RESPONSES   VCAM-1  

摘要： BackgroundActivating transcription factor 3 (ATF3) is an early response gene that is activated in response to atherosclerotic stimulation and may be an important factor in inhibiting the progression of atherosclerosis. In this study, we directly measured the expression of ATF3 and inflammatory factors in human coronary atherosclerotic plaques to examine the relationship between ATF3 expression, inflammation and structural stability in human coronary atherosclerotic *** total of 68 coronary artery specimens were collected from the autopsy group, including 36 cases of sudden death from coronary heart disease (SCD group) and 32 cases of acute death caused by mechanical injury with coronary atherosclerosis (CHD group). Twenty-two patients who had no coronary heart disease were collected as the control group (Con group). The histological structure of the coronary artery was observed under a light microscope after routine HE staining, and the intimal and lesion thicknesses, thickness of the fibrous cap, thickness of necrosis core, degree of lumen stenosis were assessed by image analysis software. Western blotting and immunohistochemistry were used to measure the expression and distribution of ATF3, inflammatory factors (CD45, IL-1 beta, TNF-alpha) and matrix metalloproteinase-9 (MMP-9) and vascular cell adhesion molecule 1 (VCAM1) in the coronary artery. The Pearson correlation coefficient was used to analyse the correlation between ATF3 protein expression and inflammatory factors and between ATF3 protein expression and structure-related indexes in the lesion *** with those in the control group, the intima and necrotic core in the coronary artery were thickened, the fibrous cap became thin and the degree of vascular stenosis was increased in the lesion group, while the intima and necrotic core became thicker and the fibrous cap became thinner in the SCD group than in the CHD group (P<0.05). There was no or low expression of ATF3, inflammator

关键词： DRUG-RESISTANCE   MOLECULAR-MECHANISMS   NUCLEAR-RNA   MET-DB   INFLAMMATION   METHYLATION   EXPRESSION   GENE  

摘要： Tamoxifen has been used for years for treating estrogen receptor-positive breast cancer; drug resistance, however, constitutes one of the main challenges for this therapy. We found that the protein expression level of ATF3 is significantly higher in tamoxifen-resistant (TamR) MCF-7 cells than the corresponding parental cancer cells. In addition, ATF3 protein expression is positively correlated with the resistance of TamR MCF-7 cells to 4-hydroxytamoxifen (4-OHT). Mechanistically, elevated ATF3 protein expression in TamR MCF-7 cells results from a lower level of expression of YTHDF2, an m(6)A reader protein, and the ensuing stabilization and increased translational efficiency of ATF3 mRNA. Additionally, TamR MCF-7 cells exhibited decreased methylation at A131, a consensus motif site for m(6)A, in the 5'-untranslated region (5'-UTR) of ATF3 mRNA. Moreover, augmented ATF3 stimulates the expression of ABCB1, an efflux pump that confers drug resistance in breast cancer cells, and ATF3 itself is also positively regulated by adenylate kinase 4. Together, our results uncovered a novel molecular target for m6A modification (i.e., ATF3 mRNA) and the epitranscriptomic regulator for this target (i.e., YTHDF2). We also illustrated the role of ATF3 in drug resistance, revealed its downstream target (i.e., ABCB1), and suggested ATF3 as a candidate therapeutic target for overcoming drug resistance in cancer cells.

关键词： siRNA-ATF3   转录激活因子3   乳腺丝氨酸蛋白酶抑制因子   基质金属蛋白酶2   U373MG   人胶质母细胞  

摘要： 目的构建、筛选并鉴定siRNA-ATF3表达载体,探讨转录激活因子3(ATF3)、乳腺丝氨酸蛋白酶抑制因子(Maspin)和基质金属蛋白酶2(MMP2)在转染了siRNA-ATF3表达载体的人胶质母细胞U373MG细胞株中的表达和意义。方法构建siRNA-ATF3表达载体,并转染U373MG细胞株,以荧光定量PCR和Western blot检测筛选出有效的siRNA-ATF3表达载体,以免疫细胞化学法检测转染siRNA-ATF3表达载体的U373MG细胞株中ATF3、Maspin和MMP2的蛋白表达情况。结果U373MG细胞中的ATF3蛋白表达量可被pSilencer siR1-ATF3和pSilencer siR2-ATF3表达载体有效降低。转染siRNA-ATF3表达载体后U373MG细胞生长状态差,增殖速度减慢。ATF3和MMP2在空白组和Control-siRNA组中的阳性表达量高于3个干扰组;与空白组和Control-siRNA组相比,Maspin在ATF3-siRNA组表达最高,在顺铂组和ATF3-siRNA+顺铂组中的表达降低。结论siRNA-ATF3表达载体有效抑制U373MG细胞的生长和增殖,顺铂可协同增强抑制效果,ATF3、Maspin和MMP2可能协同参与了U373MG细胞的生长和增殖,siRNA-ATF3表达载体可作为治疗抑制胶质母细胞瘤生长增殖的有效手段。

关键词： ACTIVATING TRANSCRIPTION FACTOR-3   ONE-CARBON METABOLISM   REDOX HOMEOSTASIS   GENE   STRESS   REGULATOR   SUPPORTS   DATABASE   IMPACT   FLUX  

摘要： Metabolic reprogramming is a common feature of many human cancers, including acute myeloid leukemia (AML). However, the upstream regulators that promote AML metabolic reprogramming and the benefits conferred to leukemia cells by these metabolic changes remain largely unknown. We report that the transcription factor ATF3 coordinates serine and nucleotide metabolism to maintain cell cycling, survival, and the differentiation blockade in AML. Analysis of mouse and human AML models demonstrate that ATF3 directly activates the transcription of genes encoding key enzymatic regulators of serine synthesis, one-carbon metabolism, and de novo purine and pyrimidine synthesis. Total steady-state polar metabolite and heavy isotope tracing analyses show that ATF3 inhibition reduces de novo serine synthesis, impedes the incorporation of serine-derived carbons into newly synthesized purines, and disrupts pyrimidine metabolism. Importantly, exogenous nucleotide supplementation mitigates the anti-leukemia effects of ATF3 inhibition. Together, these findings reveal the dependence of AML on ATF3-regulated serine and nucleotide metabolism.

关键词： Ischemic stroke   Middle cerebral artery occlusion   Bone marrow mesenchymal stem cells-derived   extracellular vesicles   MicroRNA-221-3p   Activating   Transcription factor 3   ACTIVATING TRANSCRIPTION FACTOR-3   INJURY   BRAIN   EXOSOMES   APOPTOSIS   RECOVERY   GROWTH  

摘要： Objective: We aim to explore the protective effect of bone marrow mesenchymal stem cells (BMSCs)-derived exosomal microRNA-221-3p (miR-221-3p) on ischemic stroke (IS) by targeting activating transcription factor 3 (ATF3). Methods: The middle cerebral artery occlusion (MCAO) mice model and oxygen-glucose deprivation (OGD) neuron model were established. Extracellular vesicles were isolated from BMSCs (BMSC-EVs) and transfected with altered miR-221-3p or ATF3 to treat the MCAO mice and OGD-treated neurons. MiR-221-3p and ATF3 expression were determined, and the contents of inflammatory factors were detected. The pathological changes and apoptosis in mice brain tissues were observed. In cellular experiments, the viability and apoptosis of OGDtreated neurons were evaluated. Binding relationship between miR-221-3p and ATF3 was determined. Results: MiR-221-3p was down-regulated and ATF3 was up-regulated in MCAO mice and OGD-treated neurons. BMSC-EVs and BMSC-EVs carrying up-regulated miR-221-3p attenuated inflammation, pathological changes and apoptosis in MCAO mice brain tissues, and also promoted viability and repressed apoptosis of OGD-treated neurons. ATF3 was verified as a target of miR-221-3p. Conclusion: BMSC-EVs carrying miR-221-3p protect against IS by inhibiting ATF3. This study may be helpful for exploring therapeutic strategies of IS.