关键词： Dorsal root ganglion   Peripheral nerve injury   Nerve regeneration   Regeneration-associated gene   Fluorescent tracer  

摘要： Peripheral nerve injury leads to deficient recovery of sensation and a causative factor may be that only 50-60% of primary sensory neurons succeed in regenerating axons after primary nerve repair. In this study, an in vivo rat sciatic nerve injury and regeneration model was combined with laser microdissection and quantitative real-time polymerase chain reaction with the aim of examining the gene expression of regenerative molecules in cutaneous and muscular sensory neurons. Recent studies have identified peripherin and ATF-3 molecules as crucial for neurite outgrowth propagation;our novel findings demonstrate a subpopulation of non-regenerating sensory neurons characterized by a failure to upregulate transcription of these molecules and that a greater peripherin mRNA expression in injured cutaneous neurons may potentiate this subpopulation to regenerate more axons than muscle afferent neurons following injury. The gene expression of the structural neurofilament NF-H is found to be significantly downregulated following injury in both sensory subpopulations. (C) 2009 Elsevier B.V. All rights reserved.

关键词： Epithelial cell replacement   Cell adhesion   Epidermis   Metamorphosis   Abdomen   bZIP  

摘要： Epithelial sheet spreading and fusion underlie important developmental processes. Well-characterized examples of such epithelial morphogenetic events have been provided by studies in Drosophila, and include embryonic dorsal closure, formation of the adult thorax and wound healing. All of these processes require the basic region-leucine zipper (bZIP) transcription factors Jun and Fos. Much less is known about morphogenesis of the fly abdomen, which involves replacement of larval epidermal cells (LECs) with adult histoblasts that divide, migrate and finally fuse to form the adult epidermis during metamorphosis. Here, we implicate Drosophila Activating transcription factor 3 (Atf3), the single ortholog of human ATF3 and JDP2 bZIP proteins, in abdominal morphogenesis. During the process of the epithelial cell replacement, transcription of the atf3 gene declines. When this downregulation is experimentally prevented, the affected LECs accumulate cell-adhesion proteins and their extrusion and replacement with histoblasts are blocked. The abnormally adhering LECs consequently obstruct the closure of the adult abdominal epithelium. This closure defect can be either mimicked and further enhanced by knockdown of the small GTPase Rho1 or, conversely, alleviated by stimulating ecdysone steroid hormone signaling. Both Rho and ecdysone pathways have been previously identified as effectors of the LEC replacement. To elicit the gain-of-function effect, Atf3 specifically requires its binding partner Jun. Our data thus identify Atf3 as a new functional partner of Drosophila Jun during development.

关键词： 肿瘤细胞转化   ATF3   P53基因   正常细胞   遗传毒性   细胞凋亡   E2F1   癌基因  

摘要： 正常细胞向肿瘤细胞转化的微环境中,细胞会碰到很多的应激信号,包括遗传毒性的损害,癌基因的激活,端粒的侵蚀,缺氧,营养剥夺等.此时细胞中一些内在的机制就会被激活从而去抑制或减少它们的损害.明显的例子就是P53基因在应答中抑制或杀死这些细胞.还有的例子就是通过癌基因的应激引起c-myc和E2F1来触发细胞凋亡.在研究肿瘤的发生发展的机制中,细胞在自我防卫和消除应激反应基因的研究扮演着重要的角色.下面就ATF3在肿瘤中的研究进展做一综述.

摘要： Activating Transcription Factor 3 (ATF3), a member of the ATF/CREB family, is induced rapidly by various stresses. Its induction mechanism and role in response to changes in cellular redox status, however, have not been elucidated. Here, we found that NF-E2-related factor 2 (Nrf2), a transcription factor known to bind to antioxidant response element (ARE) in promoters, transcriptionally upregulated ATF3 expression in astrocytes. Treatment with Nrf2 activators and oxidants provoked ATF3 induction in astrocytes, whereas its expression was reduced in Nrf2-depleted cells. We further demonstrated that the consensus ARE in the ATF3 promoter is critical for Nrf2-mediation by promoter analyses using an ATF3 promoter-driven luciferase construct and a chromatin immunoprecipitation assay. In addition, we found that Nrf2-dependent ATF3 induction contributed to the antioxidative and cytoprotective functions of Nrf2 in astrocytes. Taken together, our findings suggest that ATF3 is a new target for Nrf2 and has a cytoprotective function in astrocytes.

关键词： Biology   Biomedical Research   Oncology   ATF3   cancer   activating transcription factor   stroma   metastasis  

摘要： Breast cancer represents a very heterogeneous class of diseases originating in the mammary epithelium which are estimated to account for 1/3 of all newly diagnosed cancers in women in the U.S. this year, equating to roughly 200,000 diagnoses. In addition to its high frequency of diagnosis among all cancers in women, breast cancer is also the second leading cause of cancer-related mortality among women. Consequently, a great deal research has been devoted to understanding the various aspects of breast cancer development and progression with the hope of identifying novel therapeutic targets in order to design treatments that possess improved clinical efficacy and patient outcomes. To this end, recent advances have shed new light on the complexity of this disease. As a result, it is now accepted that in addition to the tumorigenic changes which take place in the transformed mammary epithelial cells, the microenvironment of the newly forming tumor plays a prominent role in the progression of the disease toward invasive and metastatic phenotypes. Therefore a more complete understanding necessitates investigating how breast cancer cells and components of their microenvironment reciprocally communicate and influence each other.@pqdt@break@The work presented in this dissertation is focused on the evaluation of the transcription factor ATF3, a molecule which functions as an integration point for cellular communication during changes in homeostasis and subsequent adaptation in response to those changes, in breast cancer development and metastasis. While the current literature supports the involvement of ATF3 in cancer biology, many of the reports are based on in vitro observations and are conflicting. To clarify some of these issues, we tested whether ATF3 plays a role in breast cancer development in vivo using a loss-of-function approach (chapter 2). The data presented demonstrate that ATF3 regulates both primary tumor growth and metastasis, and suggest that ATF3 expression

关键词： SOCIAL perception   ABSTRACTS  

摘要： An abstract of the article 'Variation in NMB and ATF3 protein expression in Von Economo neurons of hominoids: Implications for evolution of social cognition,' by Cheryl D. Stimpson is presented.

关键词： 肾小球系膜细胞(GMCs)   亚溶解型C5b-9(sC5b-9)   Thy-1肾炎(Thy-1N)   早期生长应答基因-1(Egr-1)   激活转录因子3(ATF3)   野生型Egr-1基因   短发夹状RNA(shRNA)   GMCs   重组质粒   Egr-1   ATF3   GMCs凋亡   基因启动子活性  

摘要： 第一部分SublyticC5b-9刺激大鼠GMCs和Thy-1肾炎大鼠肾组织Egr-1及ATF3基因表达的检查 目的:检查亚溶解型C5b-9 (sublytic C5b-9, sC5b-9)刺激大鼠肾小球系膜细胞(glomerularmesangialcells,GMCs)和Thy-1肾炎(Thy-1nephritis,Thy-1N)模型大鼠肾组织中早期生长应答基因-1(earlygrowthresponsegene-1,Egr-1)与激活转录因子3(activating transcription factor 3, ATF3)基因的表达及两者的时相关系。方法:体外培养大鼠的GMCs并给予一定剂量的sC5b-9刺激(实验设不同处理的对照组),同时应用抗大鼠胸腺细胞血清(anti-thymocyte serum, ATS)复制大鼠Thy-1肾炎模型,通过RT-PCR和Western blot检查不同分组及时间点体内外Egr-1及ATF3基因的表达情况。结果:sC5b-9刺激GMCs及大鼠Thy-1N的凋亡时相Egr-1与ATF3的表达均明显增加,且Egr-1表达于3h时到达高峰,之后逐渐下降。而ATF3的变化趋势与Egr-1相同,亦于3h达到峰值,随后开始回落。结论:体外用sC5b-9大鼠刺激GMCs和Thy-1肾炎大鼠的肾组织均可呈现Egr-1及ATF3基因的表达上调,且两个基因的表达时相基本一致。 第二部分大鼠野生型Egr-1基因及Egr-1shRNA真核表达质粒的构建与鉴定 目的:构建大鼠野生型早期生长应答基因-1(earlygrowthresponsegene-1,Egr-1)及其特异性短发夹状小干涉RNA (short hairpin RNA, shRNA)真核表达质粒,并观察两者在大鼠肾小球系膜细胞(glomerularmesangialcells,GMCs)中过表达或沉默Egr-1基因的情况。方法:用DNA重组技术将针对大鼠Egr-1基因的CDS区序列和针对其不同位点所设计的3对shRNA序列分别克隆到真核表达质粒pcDNA3.1-myc-his-A和***中。经酶切分析及序列测定正确后,用GenEscortTMⅢ转染试剂将上述两种质粒分别转染至大鼠GMCs中。随后,用Westernblot检测Egr-1蛋白的表达及筛选最佳沉默效率的shRNA。结果:限制性内切酶酶切及核酸序列分析证明,两种重组质粒均构建正确。Western blot检查表明,构建的pcDNA3.1/Egr-1质粒在大鼠GMCs中能够表达,且Egr-1 shRNA-2具有最佳的沉默效率。结论:成功构建了大鼠野生型Egr-1基因和其特异性的shRNA真核表达质粒,为进一步研究Egr-1基因的生物学功能提供了必要的实验工具。 第三部分sC5b-9刺激GMCs诱导Egr-1与ATF3基因表达关系的研究 目的:探讨过表达或沉默Egr-1对sC5b-9刺激大鼠GMCs诱导ATF3表达和GMCs凋亡的影响,并进一步确证Egr-1启动ATF3基因转录及与ATF3基因启动子的结合情况。方法:首先通过过表达及沉默Egr-1基因,用流式细胞术分析实验各组GMCs的凋亡情况;之后利用RT-PCR和Westernblot检测过表达或沉默Egr-1基因对sC5b-9刺激GMCs诱导ATF3表达的影响;另又采用荧光素酶报告基因检查Egr-1对ATF3基因启动子活性的调节;最后用ChIP实验验证了Egr-1与ATF3基因启动子区调节元件的特异性结合情况。结果:(1)在sC5b-9诱导GMCs凋亡的过程中,过表达Egr-1可引起ATF3表达上调,进而促进GMCs凋亡;而沉默Egr-1基因则能下调ATF3的表达并减轻GMCs的凋亡病变。(2)荧光素酶报告基因实验表明,Egr-1可直接上调ATF3基因启动子的活性;ChIP实验证实,Egr-1可与ATF3基因启动子结合。结论:在sC5b-9作为刺激原诱发的GMCs凋亡中,核转录因子Egr-1能够与ATF3基因启动子区相应元件结合,通过诱导ATF3基因的转录与表达调控GMCs的凋亡病变。

关键词： 非小细胞肺癌   免疫组化   转录因子   临床特征   淋巴结转移  

摘要： 目的:　　 1、研究激活转录因子3(ATF3)的在非小细胞肺癌组织、癌旁正常肺组织组织、正常肺组织及炎性肺组织表达水平;2、初步探讨非小细胞肺癌中ATF3表达与临床特征的关系。　　 方法:1、免疫组化:　　 选取98例术后病理论断为非小细胞肺癌(Non-small cell lung cancer,NSCLC)的患者,于2006年01月～2009年5月在赣南医学院第一附属医院胸外科行肺癌根治术,术后病理分期在Ⅰ～Ⅲ期,其中男性52例,女性46例,年龄28～78岁,中位年龄为55.2岁,选取同一患者距离肺癌组织≥5cm切缘的正常肺组织作为对照组,术前均未行放化疗。同时分别选取正常肺组织30例及炎性肺组织30例分别进行检测。采用免疫组化法(即二步法)检测肺组织中ATF3表达水平。　　 2、RT-PCR:　　 选取20例术后病理论断为NSCLC的患者,于2009年06月～2010年3月在赣南医学院第一附属医院胸外科行肺癌根治术切除的原发性肺癌冰冻标本,术后病理分期在Ⅰ～Ⅲ期,选择癌旁肺组织作为对照,术前均未行放化疗。采用RT-PCR检测肺组织中的ATF3的表达水平。　　 结果:1、免疫组化　　 (1)ATF3在非小细胞肺癌中阳性表达率为62.24％,在癌旁组织中阳性表达率为5.10％,在正常肺组织中无表达,在炎性肺组织中表达率为10.00％。经统计学分析,ATF3在非小细胞肺癌组织表达水平高于癌旁组织、正常肺组织及炎性肺组织中表达水平,并且具有显著差异性(P<0.01)。　　 (2)非小细胞肺癌组织中ATF3表达阳性率为62.24％。其中男性为63.46％,女性为60.87％;≤60岁为68.97％,>60岁为52.50％;鳞癌组织中ATF3阳性表达率为56.52％,腺癌为75.86％;高分化肿瘤表达率为58.06％,中分化肿瘤表达率为60.00％,低分化肿瘤表达率为70.37％。经统计学分析,ATF3表达水平与非小细胞肺癌患者性别、年龄、肿瘤组织学类型和肿瘤分化程度无明显相关性(P>0.05):　　 (3)淋巴结转移情况为:N0表达率为42.86％,N1表达率为67.50％,N2表达率为82.61％;在TNM分期中:Ⅰ期表达率为45.45％,Ⅱ期为60.38％,Ⅲ期为82.61％。经统计学分析,ATF3表达与淋巴结转移(P<0.01)及TNM分期(P<0.05)有显著差异性。　　 2、RT-PCR　　 ATF3在NSCLC组中均表达,而在癌旁组表达率为25.00％。经统计学分析,ATF3mRNA在NSCLC和癌旁正常肺组织的配对值(d±s)为0.4724±0.2183,t值为-9.650,P<0.01,差异有统计学意义,结果与免疫组化一致。在癌旁组织中ATF3在N1 N2患者阳性表达率为66.67％(4/6),N0患者的阳性表达率为7.14％(1/14),采用Fisher的概率检验法得出,P=0.014,P<0.05,总体比较有显著性差异。　　 结论:　　 1、ATF3蛋白在非小细胞肺癌表达水平与癌旁组、正常肺组织组及炎性肺组织的表达水平有显著差异性;2、ATF3蛋白在非小细胞肺癌组织中表达水平与性别、年龄、肿瘤大小、肿瘤分化程度无明显相关性;3、ATF3蛋白表达在淋巴结转移情况和TNM分期中有相关性。