关键词： activating transcription factor-3 (ATF3)   inflammation   Mycoplasma pneumonia (MP)  

摘要： ObjectivesActivating transcription factor-3 (ATF3) is a key regulator of inflammatory responses. We aimed to investigate the effects and mechanisms of ATF3 on the inflammatory cytokines are induced by Mycoplasma pneumonia (MP). Study DesignRAW264.7 and mouse peritoneal macrophages were exposed to various time with or without MP infection (3, 6, 12, 24, and 48h), and detect the expression of ATF3. Adenovirus-expression of ATF3 (Ad/ATF3) or Ad/gal was transfected into cells which were exposed to MP for 48h, RT-PCR and ELISA was used to evaluate the expression and secretion of TNF-, IL-1, IL-6, and IL-18. In addition, intravenous administration Ad/ATF3 or Ad/gal into the mice, the secretion of inflammatory cytokines were detected using ELISA. ChIP assay was used to determine whether ATF3 can bind to the promoter of Early growth response protein 1 (Egr-1). Western blot was used to detect the expression of Egr-1 and Fyn. ResultsATF3 was increased at 3, 6, 12, and 24h and the highest expression levels occurs in 6h, there is no significant differences at 24 and 48h compared with 0h or CON group in RAW 264.7. Similar results were seen in mouse peritoneal macrophages. Overexpression of ATF3 resulted in the reduction of inflammatory cytokines. ChIP assay revealed that ATF3 can bind to the promoter of Egr-1. Overexpression of ATF3 inhibited the protein expression of Egr-1 and Fyn;conversely, ATF3-deficiency promoted the expression of Egr-1 and Fyn. Overexpression of Egr-1 reduced the anti-inflammatory action of ATF3. ConclusionsATF3 inhibit the expression and release of TNF-, IL-1, IL-6, and IL-18 induced by MP in vitro and in vivo, which is associated with its negative regulation of Egr-1/Fyn signaling pathway.

关键词： Ebselen   IL-6   IL-1   Cancer induced inflammation  

摘要： Background: Chronic inflammatory skin diseases are characterized by controlled proliferation of keratinocytes. Here, activating transcription factor 3 (ATF3) might play a fundamental role. In these inflammatory diseases, proliferation is controlled and only rarely leads to cancer development which can be supported by an inflammatory microenvironment. ATF3 is a dual function protein as it suppresses pro inflammatory IL-6 and IL-8, but also acts as a pro-oncogenic factor by the suppression of p53. We therefore analyzed ATF3 expression comparing myeloid cells with keratinocytes. Objective: To dissect the bi-modal role of ATF3 we pharmacologically induced ATF3 and analyzed its influence on cytokine expression and secretion in a cell type specific manner. Methods: Since inflammatory skin diseases can be treated systemically with Cyclosporin A or Dimethylfumarate we stimulated myeloid cells and primary human keratinocytes with these drugs and analyzed gene expression by quantitative real-time PCR. Cytokine secretion was measured by ELISA. Results: In the present study, we could show that ATF3 is induced in PBMCs by DMF and weakly by Ebselen, while CsA is the most prominent inducer of ATF3 in keratinocytes without enhancing HO-1 transcription. Further we could show that induction of stress by LPS treatment elevates IL-1 beta and IL-6 and weakly ATF3 transcription in PBMCs. While transcription of both cytokines is elevated, LPS treatment mediates IL-6 secretion with only little IL-1 beta secretion. Treatment with DMF dampens LPS-induced transcription. Conclusions: Taken together, our results shed light into the different carcinogenic potential of CsA and DMF, which both target ATF3. Collectively our data demonstrate that CsA strongly induces pro carcinogenic ATF3 in keratinocytes, whereas ATF3 induction by DMF in myeloid cells acts antiinflammatory. (C) 2017 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

关键词： PNS   Axon regeneration   Facial nerve   ATF3   Muscle  

摘要： Traumatic injuries to human peripheral nerves are frequently associated with damage to nerve surrounding tissues including muscles and blood vessels. Currently, most rodent models of peripheral nerve injuries (e.g., facial or sciatic nerve) employ surgical nerve transection with scissors or scalpels. However, such an isolated surgical nerve injury only mildly damages neighboring tissues and weakly activates an immune response. In order to provide a rodent nerve injury model accounting for such nerve-associated tissue damage and immune cell activation, we developed a drop tower-based facial nerve trauma model in mice. We compare nerve regeneration in this novel peripheral nerve trauma model with the established surgical nerve injury along several parameters. These include gene expression, histological and functional facial motoneuron (FMN) regeneration, facial nerve degeneration, immune cell activation and muscle damage. Regeneration-associated genes (RAGs;e.g., Atf3) were strongly induced in FMNs subjected to traumatic and surgical injury. Regeneration of FMNs and functional recovery of whisker movement were faster in traumatic versus complete surgical injury, thus cutting down experimentation time. Wallerian degeneration of distal nerve stumps was readily observed in this novel trauma injury model. Importantly, drop tower-inflicted facial nerve injury resulted in muscle damage, activation of muscle satellite cell markers (PAX7) and pronounced infiltration of immune cells to the injury site only in this model but not upon surgical nerve transection. Thus, we provide a novel rodent PNS trauma model that can be easily adopted to other PNS nerves such as the sciatic nerve. Since this nerve trauma model replicates multiple tissue damage frequently encountered in clinical routine, it will be well suited to identify molecular and cellular mechanisms of PNS nerve repair in wild-type and genetically modified rodents.

关键词： chemotherapy   metastasis   stress response   immune modulation   ATF3  

摘要： Chemotherapy is a double-edged sword. It is anticancer because of its cytotoxicity. Paradoxically, by increasing chemoresistance and cancer metastasis, it is also procancer. However, the underlying mechanisms for chemotherapy-induced procancer activities are not well understood. Here we describe the ability of paclitaxel (PTX), a frontline chemotherapeutic agent, to exacerbate metastasis in mouse models of breast cancer. We demonstrate that, despite the apparent benefit of reducing tumor size, PTX increased the circulating tumor cells in the blood and enhanced the metastatic burden at the lung. At the primary tumor, PTX increased the abundance of the tumor microenvironment of metastasis, a landmark microanatomical structure at the microvasculature where cancer cells enter the blood stream. At the metastatic lung, PTX improved the tissue microenvironment (the "soil") for cancer cells (the "seeds") to thrive;these changes include increased inflammatory monocytes and reduced cytotoxicity. Importantly, these changes in the primary tumor and the metastatic lung were all dependent on Atf3, a stress-inducible gene, in the noncancer host cells. Together, our data provide mechanistic insights into the procancer effect of chemotherapy, explaining its paradox in the context of the seed-and-soil theory. Analyses of public datasets suggest that our data may have relevance to human cancers. Thus, ATF3 in the host cells links a chemotherapeutic agent-a stressor-to immune modulation and cancer metastasis. Dampening the effect of ATF3 may improve the efficacy of chemotherapy.

关键词： ATF3   Zebrafish   Spinal cord injury   Regeneration   Inflammation  

摘要： Zebrafish is an excellent model to study the mechanisms underlying successful central nervous system (CNS) regeneration. Previous study shows that activating transcription factor 3 (ATF3) promotes neurite outgrowth and is involved in optic nerve regeneration in zebrafish. Here, we used zebrafish model to investigate the role of ATF3 in regeneration following spinal cord injury (SCI). Quantitative polymerase chain reaction (qPCR) and in situ hybridization revealed that ATF3 mRNA levels increased at 12 h and 6 d following SCI. Double labeled immunofluorescence showed that ATF3 expressed in motoneurons. Treatment of anti-sense ATF3 morpholino (MO) inhibited locomotor recovery and decreased axon regeneration of spinal cord injured zebrafish. Further, inhibition of ATF3 up-regulated the expression of inflammatory factors tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta). These data suggest that ATF3 could promote locomotor recovery and axon regrowth in zebrafish SCI model possibly by regulating inflammatory response. (C) 2017 Elsevier Inc. All rights reserved.

关键词： activating transcription factor (ATF3)   triglyceride-rich lipoproteins (TGRL)   vascular dementia (VaD)   vascular inflammation  

摘要： Elevation of blood triglycerides, primarily triglyceride-rich lipoproteins (TGRL), is an independent risk factor for atherosclerotic cardiovascular disease and Vascular Dementia (VaD). The accumulating evidence indicates that the development of atherosclerosis and VaD are linked to vascular inflammation. Previous studies suggest a relationship of blood lipids and/or lipoproteins to vascular inflammation leading to dementia. The objective of this study is to characterize human brain microvascular endothelial cells (HBMVEC) responses to postprandial TGRL and determine the pathways that have potential to increase risk for vascular dementia. To address this question, we treated HBMVEC with TGRL lipolysis products and then we used NGS-based RNA-Seq methods for transcriptional profiling experiments. Our data demonstrated up-regulation of activating transcription factor 3 (ATF3)-dependent inflammatory pathways in HBMVEC. Additionally, system biology analysis exhibited putative lipid-related targets and corresponding isoforms of ATF3 that may be implicated in hypertriglyceridemia-induced vascular inflammation in brain. Furthermore, RNA-Seq analysis determined specific isoforms of key transcriptional regulator ATF3 and stress response signaling pathway in lipolysis products-mediated brain vascular inflammation which may lead to vascular dementia. [ABSTRACT FROM AUTHOR]

关键词： triglyceride-rich lipoproteins (TGRL)   activating transcription factor (ATF3)   vascular inflammation   vascular dementia (VaD)  

摘要： Elevation of blood triglycerides , primarily triglyceride-rich lipoproteins (TGRL), is an independent risk factor for atherosclerotic cardiovascular disease and Vascular Dementia (VaD). The accumulating evidence indicates that the development of atherosclerosis and VaD are linked to vascular inflammation. Previous studies suggest a relationship of blood lipids and/or lipoproteins to vascular inflammation leading to dementia. The objective of this study is to characterize human brain microvascular endothelial cells (HBMVEC) responses to postprandial TGRL and determine the pathways that have potential to increase risk for vascular dementia. To address this question, we treated HBMVEC with TGRL lipolysis products and then we used NGS-based RNA-Seq methods for transcriptional profiling experiments. Our data demonstrated up-regulation of activating transcription factor 3 (ATF3)-dependent inflammatory pathways in HBMVEC. Additionally, system biology analysis exhibited putative lipid-related targets and corresponding isoforms of ATF3 that may be implicated in hypertriglyceridemia-induced vascular inflammation in brain. Furthermore, RNA-Seq analysis determined specific isoforms of key transcriptional regulator ATF3 and stress response signaling pathway in lipolysis products-mediated brain vascular inflammation which may lead to vascular dementia.

关键词： ATF3   LIF   Embryo adhesion   Recurrent implantation failure  

摘要： Background: A receptive endometrium is essential for maternal-embryonic molecular communication during implantation. However, the specific molecular regulatory mechanisms of the endometrial capacity remain poorly understood. Here, we examined activating transcription factor 3 (ATF3) expression in human endometria and the functional effect of ATF3 on embryo attachment in vitro. Methods: Immunohistochemistry (IHC) was used to assess the ATF3 expression patterns in human endometria. Quantitative real-time PCR (qRT-PCR), western blotting and immunofluorescence (IF) studies were applied to explore ATF3 expression in Ishikawa cells upon estrogen (E-2) and medroxyprogesterone acetate (MPA) treatment. qRT-PCR and western blotting were performed to inspect LIF (leukemia inhibitory factor) expression after enhancement or inhibition of ATF3, and a luciferase reporter assay and ChIP-PCR were used to confirm the regulatory mechanism of ATF3 to LIF. Endometrial epithelial capacity was assessed by an in vitro model of attachment of BeWo spheroids to Ishikawa cells. Western blotting was performed to compare the expression of ATF3 in endometrial samples of recurrent implantation failure (RIF) patients with that in samples from fertile women (FER) who had undergone no less than one successful embryo transplantation. Results: ATF3 was located in human endometrial epithelial cells and stromal cells and was significantly induced by E-2 and MPA in Ishikawa cells. Adenovirus-mediated overexpression of ATF3 in Ishikawa cells activated LIF promoter activity and enhanced its expression. Accordingly, the stimulation of BeWo spheroid adhesion promoted by ATF3 was inhibited by pretreatment with a specific antibody against LIF via the antibody-blocking assay. Moreover, ATF3 was aberrantly decreased in the endometria of RIF patients. Conclusions: Our findings suggest that ATF3 plays a significant role in regulating human endometrial receptivity and embryo attachment in vitro via up-regulation of

关键词： 乳腺肿瘤   免疫组织化学   激活转录因子3   成骨细胞特异性转录因子2  

摘要： 目的探讨激活转录因子3（ATF3）和成骨细胞特异性转录因子2（Runx2）在乳腺癌组织中的表达及临床意义。方法采用免疫组织化学法检测ATF3和Runx2在105例乳腺浸润性导管癌和相应癌旁乳腺组织中的表达情况，分析其与乳腺癌患者临床病理特征的关系。结果乳腺浸润性导管癌组ATF3和Runx2的阳性表达率分别为80.95%（85/105）和69.52%（73/105），显著高于癌旁乳腺组织的11.43%（12/105）和8.57%（9/105），差异均具有统计学意义（χ2=102.097，P=0.000；χ2=11.595，P=0.001）。ATF3和Runx2的表达与乳腺癌的组织学分级（χ2=14.623，P=0.001；χ2=24.891，P=0.000）、淋巴结转移（χ2=7.059，P=0.008；χ2=6.358，P=0.012）及pTNM分期相关（χ2=5.807，P=0.016；χ2=4.902，P=0.027），而与患者年龄（χ2=0.274，P=0.601；χ2=1.554，P=0.213）及肿瘤大小（χ2=2.476，P=0.290；χ2=5.261，P=0.072）无关。ATF3和Runx2在乳腺癌组织中的表达呈正相关（C=0.498，P=0.000）。结论ATF3和Runx2可能协同参与了乳腺癌的发生，而且ATF3和Runx2与乳腺癌的浸润、转移及临床分期有关，提示二者可能成为预测乳腺癌恶性程度和评估患者预后的重要指标。

关键词： Berberine   Hepatocellular carcinoma   KLF6   ATF3   p21   E2F1   PTTG1   ERK1/2  

摘要： Background: Hepatocellular carcinoma (HCC) is the seventh most common malignancy and the third leading cause of cancer-related death worldwide with an extremely grim prognosis. Berberine (BBR) has been found to inhibit proliferation of human HCC cells, although the underlying mechanism(s) are unclear. Methods: Protein expression was detected by Western blots. Cell viability was determined by using the CellTiter Assay kit. Results: We confirm that BBR treatment inhibits HepG2, Hep3B, and SNU-182 cell viability, and suggest that it regulates this proliferation via the modulation of multiple tumorigenesis-related genes protein expression. BBR treatment up-regulated protein expression of tumor suppressor genes, including Kruppel-like factor 6 (KLF6), activating transcription factor 3 (ATF3) and p21, while down-regulating the expression of selected oncogenes, including E2F transcription factor 1 (E2F1) and pituitary tumor transforming gene 1 (PTTG1). The specific extracellular signal-regulated kinases 1/2 (ERK1/2) inhibitor, PD98059, partially inhibited BBR effects including reduction of cell viability, and up-regulation of KLF6 and ATF3 expressions;although, PD98059 did not alter the down-regulation of E2F1 and PTTG1 expression by BBR. Conclusions: Our results suggest that BBR inhibits HCC cell viability by modulating multiple tumorigenesis-related genes, and that up-regulation of tumor suppressor genes by BBR is in part the result of ERK1/2 action. The results of this study augment our understanding of the mechanisms underlying the effect of BBR on hepatocellular cancers and provide further evidence as to the biological plausibility of this agent's role in the treatment of these malignancies.