关键词： Infantile hemangioma   Photodynamic therapy   Human hemangioma xenograft model   Photofrin   Ischemic endothelial cell damage   CELL-DEATH   CANCER   TISSUE   EXPRESSION   PHOTOFRIN   XENOGRAFT   PATHWAYS   HIF-1   ATF3   PDT  

摘要： The authors investigated the efficacy of photodynamic therapy against infantile hemangioma using a hemangioma animal model. Eighty-three hemangioma specimens from five children were implanted into nude mice. The gross and volume changes of the implants were evaluated for up to 13 weeks. The histological change of the implant was evaluated at 5 weeks after transplantation. Photodynamic therapy was performed between 6 and 10 weeks after transplantation. The photosensitizer uptake of the implant was evaluated at 24 h after photosensitizer administration. The implant response was evaluated at 0, 12, and 24 h after light delivery. The change in ATF3 levels, a transcription factor induced under severe hypoxic conditions, was investigated immediately after treatment. The implant volume increased slowly during the first 4 weeks and then involuted. At 5 weeks after transplantation, plump endothelial cells formed tightly packed sinusoidal channels, and the endothelial cells were positive for CD31 and GLUT1 expression. At 24 h after photosensitizer administration, confocal analysis showed that the photosensitizer was present within CD31-positive cells. The implant volume was significantly decreased in the treated implants compared with the untreated implants (p < 0.0001). At 24 h after light delivery, most cells had collapsed. ATF3 expression increased gradually and then reached a maximum level at 4 h after treatment. Photodynamic therapy was effective in the treatment of infantile hemangioma. Apoptosis, a major mechanism of hemangioma destruction in the early phase, might be caused by ischemic injury as well as direct effects of photodynamic therapy.

关键词： BCL2   MCL1   BCLXL   NOXA   Reactive oxygen species   ATF3   CANCER-CELLS   ER STRESS   APOPTOSIS   MCL-1   RESISTANCE   BCL-2   MECHANISMS   INHIBITORS   DISCOVERY   BALANCE  

摘要： S1 is a putative BH3 mimetic proposed to inhibit BCL2 and MCL1 based on cell-free assays. However, we previously demonstrated that it failed to inhibit BCL2 or induce apoptosis in chronic lymphocytic leukemia (CLL) cells, which are dependent on BCL2 for survival. In contrast, we show here that S1 rapidly increases reactive oxygen species, initiates endoplasmic reticulum stress, and upregulates the BH3-only protein NOXA. The BCL2 inhibitors, ABT-737, ABT-263, and ABT-199, have demonstrated pro-apoptotic efficacy in cell lines, while ABT-263 and ABT-199 have demonstrated efficacy in early clinical trials. Resistance to these inhibitors arises from the upregulation of anti-apoptotic factors, such as MCL1, BFL1, and BCLXL. This resistance can be induced by co-culturing CLL cells on a stromal cell line that mimics the microenvironment found in patients. Since NOXA can inhibit MCL1, BFL1, and BCLXL, we hypothesized that S1 may overcome resistance to ABT-737. Here we demonstrate that S1 induces NOXA-dependent sensitization to ABT-737 in a human promyelocytic leukemia cell line (NB4). Furthermore, S1 sensitized CLL cells to ABT-737 ex vivo, and overcame resistance to ABT-737 induced by co-culturing CLL cells with stroma.

关键词： ADRBK1   AKT3   ATF3   DIO2   rSNPs   TBXA2R and VEGFA genes   transcriptional factor binding site   transcriptional factors  

摘要： Regulatory single nucleotide polymorphisms (rSNPs) which change the transcriptional factor binding sites (TFBS) for transcriptional factors (TFs) to bind DNA were reviewed for the ADRBK1 (GRK2), AKT3, ATF3, DIO2, TBXA2R and VEGFA genes. Changes in the TFBS where TFs attach to regulate these genes may result in human sickness and disease. The highlights of this previous work were reviewed for these genes. [ABSTRACT FROM PUBLISHER]

关键词： TRANSCRIPTION FACTOR   AXONAL OUTGROWTH   SIGNAL-TRANSDUCTION   ATF3   PHOSPHORYLATION   EXPRESSION   INDUCTION   TRANSPORT   FACTOR-3   GANGLIA  

摘要： We investigated (a) if activation of the mitogen activated protein kinase (MAPK) pathway was linked to the stress activated protein kinase (SAPK) pathway and (b) if JNK was required for activation of c-Jun in Schwann cells of rat sciatic nerve following injury. To this aim, ERK1/2 and the transcription factors c-Jun and ATF-3 were studied by immunohistochemistry in segments of transected nerves. We utilized pharmacological inhibitors of both signal transduction pathways in vitro to determine the effects on downstream signalling events, such as c-Jun activation, and on Schwann cell survival and proliferation. A transection induces c-Jun and ATF-3 transcription in Schwann cells. These events are followed by Schwann cell activation of c-Jun in the injured nerve. The MAPK inhibitor U0126 blocked ERK1/2 activation and reduced Schwann cell proliferation as well as induction of c-Jun transcription. The JNK inhibitor SP600125 reduced Schwann cell proliferation, but did not affect the expression of ERK1/2 or injury-induced increases in c-Jun or ATF-3 levels. Importantly, nerve injury induces Schwann cell activation of c-Jun by phosphorylation, which, in contrast to in sensory neurons, is JNK independent. MAP kinases, other than JNK, can potentially activate c-Jun in Schwann cells following injury; information that is crucial to create new nerve reconstruction strategies.

关键词： HCT116   ATF3   慢病毒过表达   增殖   肿瘤起始能力   迁移侵袭   EMT   Carfilzomib  

摘要： 【目的】全球范围内，结直肠癌致死位居肿瘤死因第四位，尽管如今结直肠癌的诊断以及治疗水平有所提高，其5年生存率也只有53.8-65.2%。ATF3蛋白属于ATF/CREB转录因子家族成员具有碱性亮氨酸拉链结构（bZip）,作为一种转录活化因子在肿瘤中起着致癌基因还是抑癌基因作用存在争论。本研究通过慢病毒介导ATF3在结直肠癌细胞HCT116中过表达，探讨ATF3在HCT116细胞中对其增殖、肿瘤起始能力、转移侵袭以及上皮间质转化（EMT）生物学作用，并初步探讨可能的分子机制,探讨蛋白酶体抑制剂Carfilzomib对HCT116的细胞毒作用以及对ATF3的调节。 【方法】构建慢病毒介导的ATF3表达载体并感染HCT116，puro筛选出稳定ATF3过表达的HCT116细胞株HCT116/ATF3和阴性对照HCT116/Control。应用MTS细胞活性分析、FACS细胞周期检测、FACS AnexinⅤ-7-AAD、克隆形成实验来检测细胞增殖能力;应用细胞球囊形成实验以及FACS检测细胞肿瘤起始能力;应用划痕实验、Transwell转移分析和Transwell侵袭分析来检测细胞转移侵袭能力;应用光学显微镜观察细胞形态变化鉴定细胞EMT;用Western Bloting和real-time qPCR来检测相关基因的mRNA及蛋白表达情况。 【结果】MTS细胞活性检测增殖能力以及克隆形成实验克隆数在HCT116/ATF3细胞明显低于对照细胞HCT116/Contro（lp<0.05）;real-time qPCR和Western Blot检测到HCT116/ATF3细胞survivin、cyclinD1、PCNA表达下调以及PARP表达上调。HCT116/ATF3细胞形成的细胞球囊平均直径和数量以及CD133+CD24+细胞群的百分比明显低于对照细胞（p<0.05）;qRT-PCR和Western Bloting检测到HCT116/ATF3细胞CD133、 CD24、Ep-CAM、 CD166、 ID1、beta-catenin和EZH2表达下调。HCT116/ATF3细胞创伤愈合划痕实验创伤愈合程度、Transwell迁移分析与Transwell侵袭实验的细胞穿过数明显低于对照细胞（p<0.05）;real-time qPCR和Western Blot检测到HCT116/ATF3细胞MMP2和MMP9表达下调以及TIMP2和TIMP1表达上调。光学显微镜观察细胞形态HCT116/ATF3细胞形态由间质细胞向上皮细胞转化（MET）;Western Blot检测到HCT116/ATF3细胞E-cadherin表达上调和snail、slug和twist表达下调。通过MTS细胞活性检测HCT116生长随Carfilzomib剂量增加而受到抑制，细胞周期检测Carfilzomib作用后HCT116主要停滞在G0/G1期;FACS AnexinⅤ-7-AAD检测HCT116凋亡随Carfilzomib剂量增加而增加。通过real-time qPCR和Western Blot检测到Carfilzomib诱导ATF3表达具有时间依赖性。 【结论】应用慢病毒介导基因转染成功构建ATF3稳定表达HCT116细胞模型HCT116/ATF3和对照细胞HCT116/Control;ATF3通过下调survivin、cyclinD1、PCNA以及上调PARP来抑制HCT116细胞增殖;ATF3通过下调CD133、 CD24、Ep-CAM、 CD166、 ID1、beta-catenin和EZH2来抑制HCT116肿瘤起始能力;ATF3通过下调MMP2和MMP9以及上调TIMP1和TIMP2来抑制HCT116细胞迁移侵袭能力;ATF3通过上调E-cadherin和下调snail、slug和twist逆转HCT116EMT;蛋白酶体抑制剂Carfilzomib可上调HCT116中ATF3的表达并抑制HCT116细胞生长。

关键词： p53   ATF3   骨肉瘤   基因治疗  

摘要： 目的：骨肉瘤(osteosarcoma)是较常见的发生在20岁以下的青少年或儿童的一种恶性骨肿瘤,在小儿骨恶性肿瘤中最多见,约为小儿肿瘤的5%。骨肉瘤起源于间叶组织,恶性程度高,侵袭性强,较早出现肺部转移,因而骨肉瘤患者的预后非常差,患者生存率很低,截肢后5年存活率仅为5-15%。现如今分子靶向治疗的出现,给骨肉瘤患者带来了新的福音。应激反应基因ATF3是P53的一个重要靶基因,在多种癌症中具有抑癌基因的作用,ATF3基因能调控细胞死亡,细胞周期。RITA (reactivation of p53and induction of tumor cell apoptosis)是一种P53靶向小分子抗癌化合物,诱导P53及其靶基因表达,在许多细胞系中导致大量细胞凋亡。本课题研究了ATF3基因在抗癌化合物RITA诱导的骨肉瘤细胞(U20S)凋亡中起到的调节作用。 方法：不同浓度及时间长度RITA刺激骨肉瘤细胞系(U20S),通过Westerblot及荧光定量PCR测定ATF3蛋白及mRNA表达水平。将ATF3沉默载体转入骨肉瘤细胞系(U20S),建立稳定细胞系,RITA处理细胞,用MTT、流氏细胞术测定细胞凋亡水平,通过Westerblot及荧光定量PCR测定ATF3下游凋亡调控基因DR5、PUMA的蛋白及mRNA水平。 结果： 1、用0.2uM RITA处理U20S细胞后,取0,2,4,8,12,24小时收取蛋白及mRNA后,用westernblot及q-RTPCR检测ATF3蛋白及mRNA水平。结果显示：第0小时(即空白对照组)ATF3蛋白及mRNA表达水平极低,从第2小时开始ATF3蛋白及mRNA水平显著升高,而且随着时间延长ATF3蛋白及mRNA水平也随之增高,说明RITA对ATF3蛋白及mRNA水平的诱导具有时间依赖性。选取OuM,0.05uM,0.1uM,0.2uM,0.5uM,1uM RITA处理U20S细胞24小时后收取蛋白及mRNA,用westernblot及q-RTPCR检测ATF3蛋白及mRNA水平。结果显示：未用RITA处理的细胞(即空白对照组)ATF3蛋白及mRNA表达水平极低,而随着RITA浓度增加,ATF3蛋白及mRNA水平显著升高,说明RITA对ATF3蛋白及mRNA水平的诱导具有浓度依赖性。 2、 ATF3siRNA转染U20S细胞系,分为：空白对照组,空白对照组+RITA, siATF3组,siATF3+RITA组,0.2uM RITA诱导时间为24小时,westernblot结果显示ATF3siRNA转染U20S细胞后,对比空白对照组及空白对照组+RITA组,siATF3组及siATF3+RITA组均很难检测到ATF3蛋白表达。 3、用不同浓度RITA处理空白对照组及两个ATF3沉默组,MTT结果显示,RITA的不同浓度在固定的时间(72h)对骨肉瘤细胞的抑制作用存在差异,抑制作用随浓度的增大而增加;在同一浓度中,ATF3沉默组细胞较空白对照组对RITA的抵抗力显著增强。 4、流式细胞术测定细胞凋亡siATF3组对比空白对照组,细胞凋亡率差别无统计学意义,siATF3+RITA组对比空白对照组+RITA组,细胞凋亡率显著降低。 5、westernblot结果显示：siATF3组对比空白对照组,DR5、PUMA蛋白水平均显著降低,siATF3+RITA组对比空白对照组+RITA组,DR5、PUMA蛋白水平也显著降低。q-RTPCR结果显示：siATF3组对比空白对照组,DR5、PUMA mRNA水平无明显差异;siATF3+RITA组对比空白对照组·+RITA组,DR5、PUMA mRNA水平显著降低。 结论： 1、RITA诱导ATF3基因在骨肉瘤细胞U2OS中高表达。 2、骨肉瘤细胞系U2OS中ATF3沉默后RITA无法诱导ATF3蛋白表达。 3、ATF3基因沉默后骨肉瘤细胞U2OS对RITA抵抗力增强。 4、ATF3基因沉默后骨肉瘤细胞U2OS对RITA抵抗力增强是通过调控下游凋亡基因实现的。

关键词： 转录激活因子3   苔藓纤维出芽   癫痫  

摘要： 目的通过观察戊四氮(pentylenetetrazole;PTZ)大鼠点燃模型海马CA3区及齿状回区转录激活因子3(activating transcription factor3;ATF3)的动态表达变化,结合海马苔藓纤维出芽(mossy fiber sprouting;MFS)的程度,探讨其与MFS的关系及在癫痫过程中的作用。 方法健康SD雄性成年大鼠150只,随机分为生理盐水对照组(60只)和PTZ实验组(90只)。实验组每日腹腔注射PTZ (30mg/kg);同时生理盐水组注射等量生理盐水。按照首次腹腔注射后第3天,1周,2周,4周和6周的时间点分配,将PTZ组及生理盐水组随机分为5个亚组。根据实验方法的不同再将每个亚组分为三个小组：第一组用于免疫组化及免疫荧光,检测各时间点ATF3蛋白在海马各区的动态表达及在细胞内表达定位;第二组用于Western Blot,将海马各区ATF3表达变化进行半定量分析;第三组用于Timm染色,检测苔藓纤维出芽情况,其中实验组每小组各6只,对照组每小组各4只。 结果 1.动物行为学表现 大鼠多在连续注射PTZ21-28天(平均23.7±1.9天)后被点燃。模型点燃率为94.4%,死亡率为5.33%。除了4周和6周组各有一只大鼠未出现痫性发作外,点燃成功后大鼠出现诱发发作及自发发作,对照组未出现痫性发作。 2.苔藓纤维出芽评分 实验组CA3区与对照组相对应时间点相比MFS评分均有显著性差异(P<0.05)。从3天起评分逐渐增加,4周时达高峰,至6周仍维持在较高水平。齿状回区MFS评分在实验组与对照组之间以及各组内不同时间点之间比较均无显著性差异(P>0.05)。 ***3蛋白的动态表达 免疫组化示在癫痫点燃过程中,实验组：海马CA3区ATF3蛋白表达逐渐上升,在4周组达最高峰,6周组稍有下降(P<0.05);齿状回区ATF3蛋白在点燃过程中无明显变化(P>0.05)。对照组：海马CA3区及齿状回区在3天到6周组均无明显变化(P>0.05)。Western Blot结果示海马ATF3的表达自3天组逐渐上升,至4周组达高峰,6周组较前明显下降;除了3天组之外,其他各时间点ATF3表达在实验组均较对照组高(P<0.05)。免疫荧光结果示ATF3蛋白定位于大鼠海马神经元细胞核及细胞浆中,且主要集中在细胞浆。 结论 ATF3蛋白在海马CA3区的表达上调可能促进苔藓纤维出芽,进而导致颞叶癫痫的发生及发展。

关键词： Carcinoma   Hepatocellular   Activating transcription factor 3 (ATF3)   Gene expression   RESPONSE GENE   CYCLIN D1   ATF3  

摘要： Background and Aim: To explore the expression and role of activating transcription factor 3 in human hepatocellular carcinoma. Methods: Immunohistochemistry, Western blot assay and Real-time PCR were used to evaluate activating transcription factor 3 protein and gene level in HCC clinical samples. Results: Activating transcription factor 3 expression is lowest in HCC, and the protein level is lower in patients with capsule invasion, while there is no association with other main clinical pathological features. Conclusions: Low expression of ATF3 may function as a tumor suppressor during human hepatocellular oncogenesis and targeting ATF3 pathway might be beneficial for anti-HCC therapy. (C) 2014 Published by Elsevier GmbH.

关键词： 兔椎间盘退变模型   等离子汽化消融术   TNF-α   ATF3   GAP43  

摘要： 目的：研究等离子汽化消融术兔退变椎间盘内TNF-α和背根神经节内ATF3、GAP43神经元表达的影响，探索其治疗盘源性腰痛的机制;在不同椎间盘模型内的差异表达，探索盘源性腰痛发病机制。 方法：健康新西兰大白兔，不计雌雄72只，体质量2.5-3.5kg，排除受孕及患病兔，采用针刺法制造新西兰大白兔L5-6椎间盘退变模型，所有实验动物学均在不同阶段进行椎间盘内注入神经示踪剂-荧光金（Fluoro-Gold），根据椎间盘退变和等离子汽化消融术两因素进行析因分析设计分组，依据椎间盘退变因素随机分为两组A组（椎间盘退变组）和B组（正常椎间盘组），然后以等离子汽化消融术因素随机分为治疗组（A1组或B1组）对照组（A2组或B2组，假手术组），收集不同时间段的椎间盘标本和背根神经节标本，免疫组化检查椎间盘髓核内TNF-α和背根神经节内ATF3、GAP43阳性神经元数目。统计分析采用SPSS17.0软件进行,检验水准α＝0.05，P＜O.05认为差异有统计学意义。 结果：正常组治疗组术后1天椎间盘组织中TNF-α的活性为90.23±5.56，术后1周为88.45±4.37，术后2周87.33±5.84;正常组治疗组术后1天椎间盘组织中ATF3、GAP43阳性细胞计数分别为2.73±1.74、2.62±1.63，术后1周为2.54±1.51、2.51±1.49，术后2周2.28±1.27、2.25±1.20;正常组对照组术后1天TNF-α的活性为91.18±4.36，术后1周为91.78±5.25，术后2周93.33±5.45;正常组对照组术后1天ATF3、GAP43阳性细胞计数为3.05±1.44、3.02±1.35，术后1周为3.21±1.49、3.16±1.35，术后2周3.35±1.78、3.29±1.62;模型治疗组消融术后1天椎间盘组织内TNF-α活性为184.78±5.43，术后1周为154.32±4.66，术后2周为141.46±8.52;模型治疗组消融术后1天椎间盘组织内ATF3、GAP43阳性细胞计数为5.63±1.46、5.55±1.37，术后1周为5.12±1.42、5.04±1.34，术后2周为4.65±1.34、4.56±1.26;模型对照组相应时间点椎间盘组织内TNF-α活性分别为195.78土10.82，196.43±12.80，225.76±11.38;模型对照组相应时间点椎间盘组织内ATF3、GAP43阳性细胞计数分别为6.23土1.32，6.19土1.26;6.45土1.36，6.39土1.28;7.32±1.68，7.28±1.61。TNF-α活性、ATF3、GAP43阳性细胞计数:模型组各时间点均较正常组明显升高(P

关键词： ACTIVATING TRANSCRIPTION FACTOR-3   CALCIUM-CHANNELS   REDOX MODULATION   IMMUNE CELLS   NEURONS   ATF3   CONTRIBUTES   MECHANISMS   GROWTH   MODEL  

摘要： Aims: Neuropathic pain is a chronic debilitating disease that is often unresponsive to currently available treatments. Emerging lines of evidence indicate that reactive oxygen species (ROS) are required for the development and maintenance of neuropathic pain. However, little is known about endogenous mechanisms that neutralize the pain-relevant effects of ROS. In the present study, we tested whether the stress-responsive antioxidant protein Sestrin 2 (Sesn2) blocks the ROS-induced neuropathic pain processing in ***: We observed that Sesn2 mRNA and protein expression was up-regulated in peripheral nerves after spared nerve injury, a well-characterized model of neuropathic pain. Sesn2 knockout (Sesn2(-/-)) mice exhibited considerably increased late-phase neuropathic pain behavior, while their behavior in acute nociceptive and in inflammatory pain models remained unaffected. The exacerbated neuropathic pain behavior of Sesn2(-/-) mice was associated with elevated ROS levels and an enhanced activating transcription factor 3 up-regulation in sensory neurons, and it was reversed by the ROS scavenger N-tert-Butyl--phenylnitrone. In contrast, administration of the ROS donor tert-butyl hydroperoxide induced a prolonged pain behavior in naive Sesn2(-/-) mice. Innovation: We show that the antioxidant function of Sesn2 limits neuropathic pain processing in vivo. Conclusion: Sesn2 controls ROS-dependent neuropathic pain signaling after peripheral nerve injury and may, thus, provide a potential new target for the clinical management of chronic neuropathic pain conditions. Antioxid. Redox Signal. 19, 2013-2023.