关键词： ASC-J9 (R)   ATF3   Prostate cancer   ATF3 response element   PTK2   ACTIVATING TRANSCRIPTION FACTOR-3   DEGRADATION ENHANCER ASC-J9(R)   TARGETING ANDROGEN RECEPTOR   UP-REGULATION   CASTRATION   ATF3   METASTASIS   APOPTOSIS   ENZALUTAMIDE   INFLAMMATION  

摘要： BackgroundEarly studies indicated that ASC-J9 (R), an androgen receptor (AR) degradation enhancer, could suppress the prostate cancer (PCa) progression. Here we found ASC-J9 (R) could also suppress the PCa progression via an AR-independent mechanism, which might involve modulating the tumor suppressor ATF3 *** lentiviral system was used to modify gene expression in C4-2, CWR22Rv1 and PC-3 cells. Western blot and Immunohistochemistry were used to detect protein expression. MTT and Transwell assays were used to test the proliferation and invasion ***-J9 (R) can suppress PCa cell proliferation and invasion in both PCa C4-2 and CWR22Rv1 cells via altering the ATF3 expression. Further mechanistic studies reveal that ASC-J9 (R) can increase the ATF3 expression via decreasing Glutamate-cysteine ligase catalytic (GCLC) subunit expression, which can then lead to decrease the PTK2 expression. Human clinical studies further linked the ATF3 expression to the PCa progression. Preclinical studies using in vivo mouse model also proved ASC-J9 (R) could suppress AR-independent PCa cell invasion, which could be reversed after suppressing ***-J9 (R) can function via altering ATF3/PTK2 signaling to suppress the PCa progression in an AR-independent manner.

关键词： glioma   brucine   ferroptosis   ATF3   hydrogen peroxide   ER stress   NOX4  

摘要： Ferroptotic cell death is characterized by iron-dependent lipid peroxidation that is initiated by ferrous iron and H2O2 via Fenton reaction,in which the role of activating transcription factor 3 (ATF3) remains *** is a weak alkaline indole alkaloid extracted from the seeds of Strychnos nux-vomica,which has shown potent antitumor activity against various tumors,including *** this study,we showed that brucine inhibited glioma cell growth in vitro and in vivo,which was paralleled by nuclear translocation of ATF3,lipid peroxidation,and increases of iron and ***,brucine-induced lipid peroxidation was inhibited or exacerbated when intracellular iron was chelated by deferoxamine (500 μM) or improved by ferric ammonium citrate(500 μM).Suppression of lipid peroxidation with lipophilic antioxidants ferrostatin-1 (50 μM) or liproxstatin-1 (30 μM) rescued brucine-induced glioma cell ***,knockdown of ATF3 prevented brucine-induced accumulation of iron and H2O2 and glioma cell *** revealed that brucine induced ATF3 upregulation and translocation into nuclei via activation of ER ***3 promoted brucine-induced H2O2 accumulation via upregulating NOX4 and SOD1 to generate H2O2 on one hand,and downregulating catalase and xCT to prevent H2O2 degradation on the other hand.H2O2 then contributed to brucine-triggered iron increase and transferrin receptor upregulation,as well as lipid *** was further verified by treating glioma cells with exogenous H2O2 ***,H2O2 reversely exacerbated brucine-induced ER *** together,ATF3 contributes to brucine-induced glioma cell ferroptosis via increasing H2O2 and iron.

关键词： Atherosclerosis   ATF3   Macrophage   Bioinformatics analysis   PI3K-AKT pathway   DISEASE  

摘要： ATF3 (activating transcription factor 3) is a member of the mammalian activation transcription factor/cAMP-responsive element-binding (CREB) family. It plays a role in inflammation and innate immunity, and suggests that ATF3 is associated with atherosclerosis. In our study, we analyzed datasets of atherosclerosis from the NCBI-GEO (Gene Expression Omnibus) database and found that expression levels of ATF3 were lower in macrophages from ruptured atherosclerotic plaques than from stable atherosclerotic plaques. Expression levels of ATF3 correlated with the stability of atherosclerotic plaques. KEGG analysis of different expression genes (DEGs) between ruptured and stable atherosclerotic plaques was performed by Metascape database. The PI3K-AKT pathway may be a potential pathway of the formation of ruptured atherosclerotic plaques. High-fat diet-induced atherosclerosis apoE(-/-)mice were divided into two groups: a model group and an ATF3 overexpression (OE)-group. Tests on atherosclerotic plaques in the aortic root suggested that absence of ATF3 and increase of macrophages may be risk factors for the formation of ruptured atherosclerotic plaques. We found decreased areas of lesions in aortic roots and branches of aortic arch, as well as increased lesional content of macrophages as well as TUNEL-positive areas. Consistent with these results, we found reduced degradation and incidence of elastic plate cracks accompanied by suppressed MMPs expression and transduction pathway protein PI3K/AKT activation. These data suggest that ATF3 is a signaling molecule that mediates the progression and stability of atherosclerotic plaques. ATF3 could be a potential new biomarker for the prognosis of atherosclerosis and may be a therapeutic target to reduce atherosclerosis.

关键词： REVERSE CHOLESTEROL TRANSPORT   HIGH-DENSITY-LIPOPROTEIN   ACTIVATING TRANSCRIPTION FACTOR-3   DUAL-RELEASE HYDROCORTISONE   CORONARY-ARTERY-DISEASE   RECEPTOR-DEFICIENT MICE   SR-BI   ADRENAL INSUFFICIENCY   PREVENTS ATHEROSCLEROSIS   URSODEOXYCHOLIC ACID  

摘要： Activating transcription factor (ATF)3 is known to have an anti-inflammatory function, yet the role of hepatic ATF3 in lipoprotein metabolism or atherosclerosis remains unknown. Here we show that overexpression of human ATF3 in hepatocytes reduces the development of atherosclerosis in Western-diet-fed Ldlr(-/-) or Apoe(-/-) mice, whereas hepatocyte-specific ablation of Atf3 has the opposite effect. We further show that hepatic ATF3 expression is inhibited by hydrocortisone. Mechanistically, hepatocyte ATF3 enhances high-density lipoprotein (HDL) uptake, inhibits intestinal fat and cholesterol absorption and promotes macrophage reverse cholesterol transport by inducing scavenger receptor group B type 1 (SR-BI) and repressing cholesterol 12 alpha-hydroxylase (CYP8B1) in the liver through its interaction with p53 and hepatocyte nuclear factor 4 alpha, respectively. Our data demonstrate that hepatocyte ATF3 is a key regulator of HDL and bile acid metabolism and atherosclerosis. Xu et al. show that liver ATF3 expression is inhibited by hydrocortisone, thus promoting the development of atherosclerosis through effects on HDL cholesterol and bile acid metabolism.

关键词： ATF3   MIRI   凋亡   糖尿病   MAPK  

摘要： 研究背景:冠心病合并糖尿病(DM)的患者再灌注治疗后,发生心肌缺血再灌注损伤(MIRI)的程度更重,其具体机制尚不明确。本研究旨在探讨转录激活因子3(activating transcription factor-3,ATF3)对糖尿病大鼠MIRI的影响,为提高冠心病合并DM患者再灌注治疗的临床获益提供新的参考。方法:(1)细胞实验:重组腺病毒载体转染H9C2细胞72 h后,给予高糖(HG)及缺氧/复氧(HR)处理,共分为6组:HG+HR组、NG+HR组、HG+sham组、NG+MN+HR组、HG+Ad-ATF3-shRNA+HR组和HG+Ad-GFP-shRNA+HR组;荧光显微镜及细胞流式技术测定腺病毒的转染效率;CCK-8实验检测细胞活性;流式细胞仪检测H92C细胞凋亡水平;qRT-PCR检测Bcl-2、Bax水平;Westernblot检测ATF3及信号通路相关蛋白表达;(2)动物实验:雄性SD大鼠以高脂饲料喂养联合40 mg/kg链脲佐菌素(streptozotocin,STZ)腹腔注射,构建糖尿病大鼠模型;重组腺病毒载体转染3 d后,结扎冠状动脉左前降支30 min缺血,再灌注3 h,构建缺血再灌注损伤模型;动物实验共分为6组:NM+IR组、DM+IR组、DM+NS+sham组、DM+NS+IR组、DM+Ad-ATF3-shRNA+IR组和DM+Ad-GFP-shRNA+IR组;荧光显微镜观察腺病毒转染效率;TTC染色测定心肌梗死面积;HE、免疫组化染色评估心肌凋亡情况;全自动生化分析仪检测血清心肌酶CK、CK-MB、LDH;qRT-PCR检测Bcl-2、Bax的表达水平;Westernblot检测ATF3及信号通路相关蛋白表达。结果:细胞实验:(1)腺病毒成功转染H9C2细胞:荧光显微镜下,随腺病毒MOI值增加,绿色荧光蛋白表达增强。(2)HR及HG均可诱导H9C2细胞ATF3表达上调:与HG+Sham组相比,HR后ATF3表达量显著升高,与NG+HR组细胞相比,HG+HR组ATF3表达量升高更为明显,转染Ad-ATF3-shRNA后其表达显著下降(P<0.05)(3)ATF3下调可增加心肌细胞活性:与HG+Sham组相比,HG+HR组细胞活性显著降低;与NG+HR组相比,HG+HR组活力降低更为明显;转染Ad-ATF3-RNAshRNA后,心肌细胞细胞活性较Ad-GFP-shRNA组升高(P<0.05)。(4)ATF3下调可减轻HR诱导的凋亡反应:结果显示,与NG+HR相比,HG+HR组心肌细胞凋亡率增加,Bax的基因表达升高更为明显,Bcl-2的表达降低;与HG+Sham组相比,HG+HR组心肌细胞凋亡率增加,Bax的表达升高,而Bcl-2的表达降低;与HG+Ad-GFP-shRNA组相比,转染Ad-ATF3-shRNA后,心肌细胞凋亡水平下降,Bax的表达降低,而Bcl-2的表达升高(P<0.05)。(5)ATF3下调可激活MAPK信号通路:Western blot实验结果显示,HR诱导心肌细胞中MAPK信号通路相关蛋白ERK1/2的磷酸化水平降低;ATF3下调促进ERK1/2的磷酸化(P<0.05),而对JNK和p38的磷酸化水平无明显影响。动物实验:(1)成功建立糖尿病大鼠模型,共有90只大鼠随机血糖值>15 mmol/L;(2)腺病毒成功转染至大鼠心肌:腺病毒转染大鼠心肌后,可见明显绿色荧光表达;(3)与DM+Sham组相比,DM+IR组ATF3蛋白表达量升高,与NM+IR组相比,DM+IR组ATF3升高更明显,转染Ad-ATF3-shRNA后表达下降(P<0.05),(4)ATF3下调减轻大鼠MIRI:TTC染色结果显示,与NM+IR组相比,DM+IR组心肌梗死面积增加;与DM+Ad-GFP-shRNA+IR组相比,DM+Ad-ATF3-shRNA+IR组心肌梗死面积降低。HE染色提示,与DM+sham组相比,DM+IR组可见心肌纤维排列紊乱、细胞水肿、肌纤维断裂及大量炎性细胞浸润,DM+Ad-ATF3-shRNA+IR组损伤减轻。与DM+Sham组相比,DM+IR组LDH、CK和CK-MB升高,DM+IR组与NM+IR组相比,LDH、CK和CK-MB升高更明显,转染Ad-ATF3-shRNA后LDH、CK和CK-MB降低(P<0.05)。(5)ATF3下调减轻MIRI诱导的凋亡:TUNEL染色结果显示,与DM+Sham组相比,DM+IR组TNUEL阳性细胞比例增多,与NM+IR组相比,DM+IR组心肌细胞中TUNEL阳性细胞增多更明显。转染Ad-ATF3-shRNA后,TUNEL阳性细胞数减少(P<0.05)。(6)ATF3下调促进MIRI诱导的MAPK信号转导:与DM+Sham组相比,MIRI后

关键词： 动脉粥样硬化   ATF3   炎症反应   NF-κB信号通路   冠心病猝死  

摘要： 目的:转录激活因子3(Activating transcription factor 3,ATF3)是响应动脉粥样硬化(Atherosclerosis,AS)刺激而激活的早期应答基因之一,可能是抑制动脉粥样硬化进展的重要因子。本研究通过在人冠状动脉粥样硬化斑块内检测ATF3及相关因子表达,并在体内外敲低ATF3的表达,探讨ATF3在动脉粥样硬化中的作用及其与炎症反应的关系。方法:(1)人体标本实验:收集贵州医科大学法医学院/法医司法鉴定中心尸检案例冠脉组织标本90例,分为:机械性损伤致急速死亡同时患有冠心病32者(CHD组),冠心病猝死者36例(SCD组),对照组22例(Con组)。HE染色观察冠脉血管组织病理学结构改变,并测量冠脉血管内膜厚度、纤维帽厚度、坏死灶厚度及血管腔狭窄程度以评估血管结构变化;免疫荧光双标法观察斑块内ATF3和CD68的表达;免疫组织化学(IHC)及Western Blot法检测冠脉血管ATF3、炎症因子(CD45、IL-1β、TNF-α)、基质金属蛋白酶9(MMP-9)及血管细胞粘附分子(VCAM1)蛋白的表达及分布;Pearson相关系数法分析病变组ATF3蛋白表达与结构相关指标,ATF3蛋白表达与炎症因子间相关性,探讨ATF3基因与动脉粥样硬化斑块内炎症反应及其结构稳定性间的关系。(2)动物实验:将32只8-9周龄SPF级载脂蛋白基因敲除(Apo E)小鼠分为Con组、AS组、AAV9-e GFP及AAV9-ATF3组,每组8只,适应喂养一周后,Con组予以普通饲料,其余各组小鼠均予以高脂饲料。所有小鼠继续饲养12周后,AAV9-ATF3基因敲除组尾静脉注射5×10vg/100μl/只的AAV9-ATF3病毒,AAV9-e GFP组尾静脉注射5×10vg/100μl/只的AAV9-e GFP空载病毒,Con组及AS组100μl/只生理盐水尾静脉注射。Con组予以普通饲料喂养,其余各组高脂喂养,所有小鼠继续喂养5周后麻醉处死,取小鼠胸主动脉近主动脉弓处供病理切片用,剩余的小鼠主动脉弓、胸主动脉、腹主动脉至肾动脉分叉处的血管组织-80℃冻存供蛋白提取。小鼠主动脉大体及横断面油红O染色检测小鼠主动脉内膜脂质沉积;HE染色观测小鼠主动脉斑块面积改变;荧光显微镜观察AAV9携带的e GFP转染小鼠主动脉斑块情况;免疫组织化学及Western Blot法检测小鼠主动脉ATF3、炎症相关因子(CD45、CD68、IL-1β、TNF-α和VCAM1、MMP-2、MMP-9)、NF-κB信号通路相关因子(IKKβ、p-IKKα/β(Ser176/180)、NF-κB P65及p-NF-κB P65(Ser536))蛋白表达变化。(3)细胞实验:采用siRNA干扰,靶向敲低THP-1细胞ATF3表达,160nmo1/ml佛波酯(PMA)诱导THP-1细胞成为巨噬细胞,并进一步用80ng/L氧化低密度脂蛋白(Ox-LDL)诱导建立泡沫细胞模型。油红O染色鉴定泡沫细胞脂质沉积;Western Blot法检测ATF3基因敲低效率、NF-κB信号通路相关因子(IKKβ、p-IKKα/β(Ser176/180)、NF-κB P65及p-NF-κB P65(Ser536))蛋白表达变化。结果:(1)人体冠脉实验:从Con组、CHD组到SCD组,冠脉血管内膜厚度和坏死灶厚度逐渐增厚,管腔狭窄程度增加(P<0.05),而SCD组纤维帽较CHD组变薄(P<0.05);免疫组织化学结果显示,在Con组冠脉血管内膜炎症因子(CD45、IL-1β及TNF-α)、VCAM1和MMP-9几乎未见表达,而在CHD组和SCD组AS斑块内,上述蛋白表达水平增高(P<0.05),且SCD组较CHD组更高(P<0.05),Western Blot也证明了上述结果(P<0.05);免疫荧光结果显示,在AS斑块内,ATF3和CD68呈现部分共表达;免疫组化及Western Blot还显示,与Con组相比,CHD组和SCD组ATF3表达水平增高(P<0.01),而SCD组中ATF3蛋白表达反而低于CHD组(P<0.01);相关性分析结果显示,CHD组和SCD组AS病灶内ATF3蛋白水平与冠状动脉内膜厚度、坏死核心厚度呈负相关(P<0.01),与纤维帽厚度呈正相关(P<0.01),而与管腔狭窄程度无相关关系(P>0.05);CHD组和SCD组AS斑块内ATF3蛋白水平与炎症因子(CD45、IL-1β及TNF-α)、MMP-9及VCAM1蛋白表达分别呈负相关关系(P<0.01)。(2)动物实验:冰冻切片荧光显微镜观察小鼠主动脉斑块,AAV9携带的e GFP呈不均匀广泛分布的绿色荧光表达,转染率约8%,这提示AAV9成功转染小鼠主动脉;IHC和Western Bl

关键词： 砷中毒   支气管上皮细胞   恶性转化   转录因子  

摘要： 砷作为一种广泛存在自然界于的化合物，具有双重作用，即可致癌又可抑癌，肺作为砷毒性作用的靶器官之一，在地方砷等一些砷污染严重的地区，肺癌发病率极高;目前砷引起肺部病变的机理较复杂，还未被完全阐明;关于砷化物在诱导支气管上皮细胞发生恶性转化的研究内容还不完善，而转录因子ATF3在在肿瘤的形成与发展过程中发挥着重要作用，但在肺癌中的相关机制尚未研究阐明。　　目的：建立砷暴露和ATF3敲除支气管上皮细胞恶性转化模型，明确ATF3在砷致恶性转化中的作用及与恶性程度的相关性，并探讨分子机制，为全面了解砷对机体的影响提供理论基础，为寻找砷中毒早期生物学标志和防治新方法提供新思路。　　方法：（1）将人支气管上皮（BEAS-2B）细胞（WT）和ATF3敲除BEAS-2B细胞（ATF3KO）在含0.25μM NaAsO2的培养基中连续培养六个月，在恶性转化的不同阶段（2、4、6个月）检测ATF3表达情况及软琼脂克隆形成能力，并将砷暴露六个月后发生恶性转化的细胞进行裸鼠成瘤实验;（2）通过MTT实验、平板克隆形成实验、细胞周期检测、划痕实验、Transwell检测、细胞凋亡实验、real time RT-PCR、Western Blot等，比较四组细胞，即WT、ATF3KO、恶性转化的WT（WT-AsT）和恶性转化的ATF3KO（ATF3KO-AsT）细胞的增殖能力、凋亡抵抗能力、迁移能力和EMT情况。（3）提取WT和WT-AsT细胞的总RNA，利用真核有参转录组测序技术，寻找参与砷致恶性转化的通路。后通过real time RT-PCR、Western Blot、ELISA等，检测这些通路在四种细胞中的激活情况。　　结果：（1）长期低剂量砷暴露会增加ATF3表达，但ATF3缺失促进砷致支气管上皮细胞发生恶性转化。（2）ATF3敲除的恶性转化细胞具有更强的增殖能力、凋亡抵抗能力及迁移能力。（3）炎症信号通路、PI3K/AKT通路、JAK/STAT3通路等在砷致恶性转化过程发生了显著变化，而ATF3敲除的恶性转化细胞炎症因子IL-6、IL-8等表达升高，AKT和STAT3的磷酸化水平升高。　　结论：ATF3在砷致恶性转化中起到抑制作用，且与恶性转化细胞的恶性程度呈负相关。ATF3可通过减轻长期砷暴露过程中的慢性炎症水平以及下游的PI3K/AKT和JAK/STAT3信号通路的激活，起到抑制肺癌癌变的作用。

关键词： 铁死亡   ATF3   内质网应激   NOX4   胶质瘤  

摘要： 研究背景及目的:铁死亡(Ferroptosis)是一种新提出的程序性细胞死亡方式,这种死亡形式与神经退行性疾病、癌症、缺血再灌注损伤、脑外伤和脑出血等多种病理过程密切相关。铁死亡在形态学方面的特征是线粒体膜密度浓缩,体积变小。在生化方面的特点是由于铁的吸收、储存和输出不平衡导致的细胞内铁离子的积累。细胞内铁离子水平主要受转铁蛋白受体调节,转铁蛋白受体负责将细胞外的铁-转铁蛋白复合物通过clathrin介导的内吞作用运送到细胞内,细胞内的铁以铁蛋白的形式进行储存,而铁转运蛋白则负责铁离子向胞外输出。异常增多的铁离子能够使膜磷脂中的多不饱和脂肪酸过氧化,因而破坏细胞膜的完整性。这主要通过两种途径实现,一是作为非血红素含铁脂氧合酶的辅助因子参与酶促反应,另一种则是与过氧化氢发生芬顿反应,生成具有强过氧化能力的羟自由基。诱导铁死亡可以有效地清除前列腺癌、结直肠癌、肝癌甚至对顺铂耐药的肺癌细胞和胶质瘤干细胞等各种类型的恶性肿瘤细胞。因此,铁死亡诱导剂作为胶质瘤的治疗药物具有相当的潜力。转录激活因子3(ATF3)是属于ATF/CREB家族的具有亮氨酸拉链结构转录因子,在人胶质瘤中过度表达,并且其表达水平与肿瘤的恶性程度呈正相关性。ATF3在调控癌细胞死亡中起着双重作用。一方面,它能够抑制肝癌细胞的肿瘤发生和进展,抑制胆管癌细胞的上皮间质化转变,限制了人结直肠癌细胞的侵袭和迁移。另一方面,它可以通过改善AKT的磷酸化来保护乳腺癌细胞免受放疗的影响。ATF3在铁死亡过程中以及在细胞凋亡和坏死中均被上调。有研究发现,ATF3通过抑制x CT的表达,促进了纤维肉瘤细胞和视网膜色素上皮细胞铁死亡的发生。然而,ATF3在调控胶质瘤细胞铁死亡过程中的作用机制尚不明确。马钱子碱是从马钱子种子中提取的弱碱性吲哚生物碱,通常用于缓解关节炎和创伤性疼痛。最近的研究显示,马钱子碱对结肠腺癌、肝癌、多发性骨髓瘤、乳腺癌和胶质瘤等多种癌症具有强效的抗肿瘤活性。其抗肿瘤作用与诱导细胞凋亡和细胞周期阻滞、JNK激活、抑制血管生成和上皮间质化转变、抑制肿瘤细胞迁移和转移相关。然而,铁死亡和ATF3是否都参与马钱子碱诱导的胶质瘤细胞死亡仍不清除。因此,本研究旨在探讨铁死亡和ATF3在马钱子碱诱导的胶质瘤细胞死亡中的作用。研究方法:1、MTT法检测马钱子碱对人胶质瘤各细胞系细胞的增殖能力抑制程度;LDH释放试验检测马钱子碱诱导的胶质瘤细胞死亡程度以及各抑制剂对马钱子碱诱导细胞死亡的影响;2、对细胞内游离二价铁离子浓度测定,采用细胞亚铁离子检测试剂盒,对不同浓度马钱子碱在给定作用时间条件下诱导人胶质瘤细胞内二价游离铁离子浓度的变化情况;检测各抑制剂对马钱子碱导致的人胶质瘤细胞亚铁离子浓度变化的影响。3、Western Blot检测马钱子碱对胶质瘤细胞作用不同时间导致的相应蛋白表达情况,以及基因沉默ATF3及NOX4后马钱子碱对胶质瘤细胞作用样品中相关蛋白含量变化的影响。4、通过MDA含量测定,研究马钱子碱诱导人胶质瘤细胞脂质氧化程度及DFO、FAC、Fer-1、Lip-1、4-PBA、GSH等抑制剂对马钱子碱诱导的人胶质瘤胶质瘤脂质氧化程度的影响。5、通过H2O2浓度测量,检测马钱子碱诱导人胶质瘤细胞活性氧自由基产生情况,以及DFO等抑制剂对马钱子碱诱导人胶质瘤细胞活性氧产生的影响。6、通过NOX氧化酶检测马钱子碱诱导胶质瘤氧化应激应对机制的影响;以及DFO等抑制剂对马钱子碱诱导人胶质瘤抗氧化应激的活性的影响。7、通过siRNA沉默ATF3、NOX4研究其对马钱子碱诱导胶质瘤细胞铁死亡过程的影响。8、通过裸鼠荷瘤实验接种小鼠胶质瘤转移瘤模型,腹腔注射马钱子碱,观察马钱子碱对胶质瘤体内模型的作用;瘤体组织检测相关亚铁离子浓度、氧化应激相关指标和相应蛋白表达情况。结果:1、马钱子碱能够抑制人胶质瘤细胞的增殖能力并且诱导细胞铁死亡。2、马钱子碱能够提高人胶质瘤细胞内亚铁离子浓度和脂质过氧化程度。3、ATF3参与马钱子碱诱导胶质瘤细胞铁死亡过程4、ATF3参与并促进马钱子碱诱导胶质瘤细胞内过氧化氢过度累积。5、ATF3通过激活NOX4和抑制xCT表达促进过氧化氢生成。6、马钱子碱诱导胶质瘤细胞内质网应激并建立内质网应激-过氧化氢累积正反馈环。7、荷裸鼠显示马钱子碱能够抑制体内胶质瘤细胞增殖,并伴有肿瘤组织内亚铁离子,MDA,H2O2水平均提高、内质网应激以及半胱氨酸和GSH消耗。结论:1、马钱子碱在体内和体外均能抑制胶质瘤增殖并诱导胶质瘤细胞死亡。2、马钱子碱诱导胶质瘤细胞发生死亡过程中伴随了细胞内铁离子浓度的增加和脂质过氧化程度上升。3、马钱子碱通过ATF3调控胶质瘤细胞铁死亡。4、马钱子碱通过激活胶质瘤细胞ATF3调控细胞内亚铁离子浓度和过氧化氢含量。5、马钱子碱通过增加过氧化氢水

摘要： Cancer is a systemic disease able to reprogram the bone marrow (BM) niche towards a pro-tumorigenic state. The timing in which BM cells start to sense the development of distant tumours and the molecular players involved in cancer-adapted immune response are largely unknown. In this context, by using the MMTV-NeuT mammary carcinoma mouse model, we have recently described the BM hematopoietic and stromal changes that support the initial steps of mammary transformation and identified the Activating transcription factor 3 (Atf3) among the genes significantly up modulated during the transcriptional reprogramming of the BM hematopoietic niche, at both early and late stages of disease. Additionally, Atf3 expression in the BM perfectly correlated with the pathologic disease score. These data supported the need to further investigate the role of Atf3 during the BM emergency haematopoiesis associated with tumour progression. In order to identify in which specific BM cell populations Atf3 was more expressed, we checked ATF3 activation in different progenitor and mature immune cell subsets starting from early stages of mammary gland transformation and along tumour progression. Taking advantage of two mouse models of luminal breast cancer, we demonstrated, extending previous observations, that ATF3 induction during cancer progression is a multistep process characterized by its activation in two different subsets of BM myeloid cells. At early time point we observed high ATF3 expression and nuclear localization in common myeloid progenitor cells (FcγR+) localized in proximity of the vascular niche. This feature was maintained at invasive stage of mammary transformation, when ATF3 became also expressed in mature monocyte/macrophage cells. Of note, the nuclear localization of ATF3 was associated with the formation of myeloid progenitor clusters in the BM of tumour-bearing mice and its overexpression in progenitor cells drove the expansion and the differentiation of monocyte/macroph

关键词： 乳腺癌   ATF3   预后  

摘要： 目的:探讨ATF3在乳腺癌组织中的表达及其临床意义。方法:选取2016年1月至2016年12月经由病理学确诊并行乳腺癌根治术的66例患者的肿瘤组织和癌旁组织标本的新鲜标本以及病理组织学切片并收集患者临床病理资料。采用RT-q PCR方法检测乳腺癌及癌旁组织中ATF3 m RNA的相对表达量;采用免疫组织化学法检测ATF3在乳腺癌组织和癌旁组织中的表达情况,应用Spearman相关系数及Cox回归模型分析ATF3表达水平与临床病理因素及预后之间的关系。结果:RT-q PCR结果显示:与癌旁组织相比,乳腺癌组织中ATF3 m RNA相对表达量明显升高。与癌旁组织相比,ATF3在乳腺癌组织的mi RNA表达量为癌旁乳腺组织的2.773倍,差异有统计学意义(P<0.05)。免疫组化结果表明:ATF3在乳腺癌组织中表达明显高于癌旁组织;Spearman相关分析显示ATF3高表达与乳腺癌的临床病理分期、淋巴结转移、组织学分级、Ki67表达分呈正相关,与HER-2表达、P53表达无相关性;Cox单因素及多因素回归分析模型显示ATF3高表达是决定乳腺癌患者预后的独立风险因素。结论:乳腺癌组织中转录激活因子3(ATF3)的表达明显高于癌旁组织且ATF3高表达与乳腺癌的淋巴结转移、临床病理分期、组织学分级、Ki67表达分呈正相关,与HER-2表达、P53表达无相关性。ATF3高表达是决定乳腺癌患者预后的独立风险因素。