关键词： Taxus chinensis var   mairei   NCI-1975 cells   Apoptosis   ATF3   Hippo-YAP pathway  

摘要： Taxus chinensis var. mairei (TC) is a traditional Chinese ornamental and medicinal plant, the leaves and twigs of which are used in anti-tumor therapy in southern China. However, the mechanism and role of aqueous extract of TC (AETC) in promoting apoptosis in non-small cell lung cancer (NSCLC) cell lines has remained unclear. In this research, we observed that AETC inhibited the suppression of the proliferation of NSCLC cells and highly inhibited the proliferation of NCI-1975 cells. Furthermore, AETC exerted minimal inhibitory effects on normal human lung epithelial cells and induced apoptosis in NCI-1975 and A549 cells. The findings of RNA sequencing, qRT-PCR, western blotting, and immunofluorescence showed that upregulated ATF3 expression and ATF3 gene knockdown, respectively, increased and decreased the anti-tumor effects of AETC associated with Hippo pathway inhibition and decreased YAP degradation. Furthermore, AETC reduced the tumor volume and weight in nude mice;upregulated ATF3, p-MOB1, and p-YAP (Ser397);and actively regulated cleaved PARP and cleaved caspase-9/8/3. These findings suggest that AETC induced NSCLC cell apoptosis via the ATF3-Hippo-YAP pathway in vivo and in vitro. We also found that AETC is non-toxic to normal cells and nude mice. Thus, AETC might represent a promising adjuvant for anti-tumor therapy against NSCLC.

关键词： Macrophage senescence   Pseudomonas aeruginosa   ATF3   ROS   Inflammation  

摘要： Pseudomonas aeruginosa (PA) is one of the most prevalent pathogens that cause nosocomial infection in critical patients. Previously, we reported PA induced macrophage to senescence under the circumstance of infection. As an oxidative stress responsiveness element, activating transcription factor 3 (ATF3) might be involved in the macrophage senescence process. To test this presumption, we manipulated the expression of ATF3 in macrophage by using a PAO1 infection system. In the present study, ATF3 expression in macrophage was increased, following the duration and colony counts of PAO1 infection. Knockdown of ATF3 in macrophage resulted in increased percentage of senescent macrophage under PAO1 infection, while overexpressing ATF3 partly blocked PAO1-induced macrophage senescence. In accordance with the senescent phenotype, elevated reactive oxygen species (ROS) production was shown in ATF3 knockdown macrophages. Also, capacity of phagocytosis was also affected by manipulation of ATF3 expression in macrophages, and increased phagocytosed fluorescent beads was found in ATF3 knockdown macrophage. ATF3 might regulate the senescence process through influence on NF-kappa B translocation. During infection, the overexpression or downregulation of ATF3 in macrophage negatively modulated the translocation of NF-kappa B p65 and its phosphorylation at Ser-536. As a result, IL-6 and TNF alpha was elevated, while IL-10 decreased in case of ATF3 knockdown. In conclusion, ATF3 negatively regulates NF-kappa B translocation and activation, and participates in PA-induced macrophage senescence. As oxidative stress and inflammation induced element, ATF3 may modulate macrophage-related host defense.

关键词： Infertility   Medical research  

摘要： Decidualization is a complex process involving cellular proliferation and differentiation of the endometrial stroma and is required to establish and support pregnancy. Dysregulated decidualization has been reported to be a critical cause of recurrent implantation failure (RIF). In this study, we found that Activating transcription factor 3 (ATF3) expression was significantly downregulated in the endometrium of RIF patients. Knockdown of ATF3 in human endometrium stromal cells (hESCs) hampers decidualization, while overexpression could trigger the expression of decidual marker genes, and ameliorate the decidualization of hESCs from RIF patients. Mechanistically, ATF3 promotes decidualization by upregulating FOXO1 via suppressing miR-135b expression. In addition, the endometrium of RIF patients was hyperproliferative, while overexpression of ATF3 inhibited the proliferation of hESCs through CDKN1A. These data demonstrate the critical roles of endometrial ATF3 in regulating decidualization and proliferation, and dysregulation of ATF3 in the endometrium may be a novel cause of RIF and therefore represent a potential therapeutic target for RIF.

关键词： Male breast carcinoma   gynecomastia   immunohistochemistry   molecular markers   prognosis  

摘要： BACKGROUND: Male breast carcinoma (male BC) is an uncommon neoplasia without individualized strategies for diagnosis and therapeutics. Low overall survival (OS) rates have been reported, mostly associated with patients' advanced stage and older age. Intratumoral heterogeneity versus homogeneity of malignant epithelial cells seems to be an important factor to consider for the development of combination therapies with curative intention. OBJECTIVE: In this preliminary study, we aim to provide valuable insight into the distinct clinicopathologic features of male BC. MATERIAL AND METHODS: In a series of 40 male BC patients. we evaluated by immunohistochemistry androgen receptor: activating transcription factor 3 (ATF3);p16. cyclin D1;fatty acid synthase (FASN): fatty acid transport protein 1 (FATP1);beta 1, beta 3, beta 4, and beta 6 integrins;collagen I and collagen IV;and their interactions. Kaplan-Meier survival curves and log-rank tests were assessed for statistical analysis. RESULTS: Homogeneous epithelial staining of p16, ATF3, beta 6 integrin, FASN, and FATP1 was found to be significantly intercorrelated, and associated with high Ki67. These markers also stained tumor stromal fibroblasts. The prognostic analysis showed statistically significant associations of FASN with disease-free survival (DFS) and OS, as well as of ATF3 with OS and collagen IV with DFS. CONCLUSIONS: This study highlights, as a novel finding, the relevance of FASN. ATF3. and collagen IV immunophenotypes. which may have innovative application in the clinical management of male BC.

关键词： 绵羊   ATF3基因   生物信息学分析   营养应激   调控元件  

摘要： 本研究旨在探究绵羊ATF3基因的蛋白特征、转录调控元件及其在营养剥夺应激下的表达变化。运用生物信息学相关软件对绵羊ATF3蛋白特征和ATF3基因启动子区元件进行分析预测,并在绵羊禁食3 d后检测骨骼肌中ATF3mRNA的表达量。结果显示,绵羊ATF3氨基酸数为187,分子式为C920H1521N273O282S12,等电点为8.89,不稳定系数是71.73,总平均疏水性值为-0.630;ATF3蛋白定位于细胞核,二级结构主要由α螺旋和无规则卷曲组成,三级结构与二级结构保持一致且可信度高;进化树显示绵羊ATF3与牛、山羊、猪的同源性较高;qRT-PCR结果显示,与正常组相比,禁食组绵羊骨骼肌中ATF3mRNA表达量极显著降低;绵羊ATF3基因启动子区存在CREB、NFKB、AP1和GRE等与应激有关的转录因子结合元件,暗示了ATF3表达量下调的原因可能与这些元件和相应转录因子的结合有关。

关键词： Mesenchymal stem cells-derived extracellular vesicles   microRNA-222   ATF3   AKT1   Colorectal cancer  

摘要： BackgroundImmunotherapy has been recently established as a new direction for the treatment of colorectal cancer (CRC), a gastrointestinal cancer. In this investigation, we aimed to expound how the posttranscriptional regulation modulated by microRNA-222 (miR-222) from mesenchymal stem cells-derived extracellular vesicles (MSC-EVs) affected the AKT pathway and the immune escape in *** cell malignant phenotype, including proliferation, migration, invasion, and apoptosis, was firstly detected after co-culture with MSC-EVs. miRNAs with differential changes in CRC cells before and after EVs treatment were filtered by microarray analysis. miR-222 was then downregulated to examine its role in CRC cells in response to EVs. Cells were implanted in mice to induce xenograft tumors, and infiltrating T cells was assessed by immunohistochemistry. The mRNA microarray was used to screen target genes, followed by rescue experiments. ChIP and western blot were conducted to validate the downstream biomolecule of *** treatment of CRC cells with MSC-EVs, the expression of miR-222 was upregulated, and cell activity was increased. Inhibition of miR-222 decreased CRC malignant aggressiveness in vitro and reduced tumorigenesis and immune escape in vivo. miR-222 targeted and bound to ATF3. Downregulation of ATF3 enhanced CRC cell malignant aggressiveness, tumorigenic capacity and immune escape. Mechanistically, ATF3 inhibited AKT1 transcription and mediated the AKT ***-EVs carry miR-222 to promote CRC cell malignant aggressiveness and immune escape. miR-222 targets and binds to ATF3, which inhibits AKT1 transcriptional activity and thereby mediates the AKT pathway.

关键词： 急性肾损伤   早期诊断   体外循环   ATF3   Kim-1  

摘要： 目的:探讨尿转录活化因子3(activating transcription factor 3,ATF3)和肾损伤因子1(kidney injury molecule 1,Kim-1)对体外循环(cardiopulmonary bypass,CPB)心脏手术后急性肾损伤(acute kidney injury,AKI)早期诊断的价值。方法:选取2018年1月—2019年12月于上海市第十人民医院心脏外科行体外循环心脏手术的患者83例,根据改善全球肾脏疾病预后组织诊断标准分为AKI组和非AKI组。收集患者术前及术后2 h、6 h、12 h、24 h、48 h尿液样本,采用酶联免疫吸附试验检测各时间点尿ATF3和Kim-1的水平,并计算尿[ATF3]·[Kim-1]的值。通过绘制受试者工作特征(ROC)曲线计算曲线下面积(AUC),评估尿ATF3、Kim-1以及[ATF3]·[Kim-1]早期诊断AKI的临床价值。结果:共42例患者发生AKI。与非AKI组相比,AKI组患者术后6 h、12 h尿ATF3明显升高,且术后12 h尿ATF3水平诊断AKI的AUC为0.691(95%CI:0.576~0.807),截断值为1216 pg/mL时,灵敏度为0.43,特异度为0.85。尿[ATF3]·[Kim-1]联合检测的AUC在术后6 h为0.712,达到最高值,灵敏度和特异度分别为0.57和0.78。结论:尿ATF3在体外循环心脏术后患者的尿液中早期表达,可以作为成人体外循环术后AKI的诊断标志物。联合使用的预测价值高于单个检测,是增加诊断准确度的可行办法。

关键词： arsenic   ATF3   Bcl-xL   DR5   apoptosis  

摘要： Arsenic is one of the most common environmental pollutants eliciting serious public health issues;however, it is also a well-recognized chemotherapeutic agent for acute promyelocytic leukemia. The association between arsenic exposure and lung diseases has been established, but underlying molecular mechanisms are poorly defined. Here we investigated the toxicology of arsenic in airway epithelium. Arsenic rapidly induced the activating transcription factor ATF3 expression through the JNK and p38 pathways. The ATF3-deficient BEAS-2B cells were relatively resistant to apoptosis upon arsenic exposure, indicating a facilitatory role of ATF3 in arsenic-induced apoptosis. We further showed that ATF3 oppositely regulated the transcription of death receptor (DR5) and Bcl2-like 1 (Bcl-xL) by directly binding to the promoter DR5 and Bcl-xL. Altogether, our findings establish ATF3 as a pro-apoptotic protein in arsenic-induced airway epithelial apoptosis through transcriptionally regulating DR5 and Bcl-xL, highlighting the potential of ATF3 as an early and sensitive biomarker for arsenic-caused lung injury.

关键词： AP‐   1   ATF3   chromatin accessibility   IARs   DNA methylation   heterochromatin   DARs   senescence  

摘要： Chromatin organization and transcriptional profiles undergo tremendous reordering during senescence. However, uncovering the regulatory mechanisms between chromatin reconstruction and gene expression in senescence has been elusive. Here, we depicted the landscapes of both chromatin accessibility and gene expression to reveal gene regulatory networks in human umbilical vein endothelial cell (HUVEC) senescence and found that chromatin accessibilities are redistributed during senescence. Particularly, the intergenic chromatin was massively shifted with the increased accessibility regions (IARs) or decreased accessibility regions (DARs), which were mainly enhancer elements. We defined AP-1 transcription factor family as being responsible for driving chromatin accessibility reconstruction in IARs, where low DNA methylation improved binding affinity of AP-1 and further increased the chromatin accessibility. Among AP-1 transcription factors, we confirmed ATF3 was critical to reconstruct chromatin accessibility to promote cellular senescence. Our results described a dynamic landscape of chromatin accessibility whose remodeling contributes to the senescence program, we identified that AP-1 was capable of reorganizing the chromatin accessibility profile to regulate senescence.

关键词： ATF3   Migration   Invasion   Stemness   Epithelial mesenchymal transition   Gastric cancer cell line   Mouse  

摘要： Background Gastric cancer (GC) is one of the most common cancers and the third leading cause of cancer related mortality worldwide. The 5-year survival rate is rather low owing to advanced unresectable and distant metastasis. The EMT has been widely implicated in the stemness, metastatic dormancy, and chemoresistance of different solid tumors. Given the fact that activating transcription factor-3 (ATF3) is a member of the ATF/CREB family of transcription factors and its role in regulation of GC recurrence and metastasis remain poorly understood, the aim of the present study was to investigate its potential impact in epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) properties and GC aggression. Methods To elucidate the potential role of ATF3 in gastric cancer, we utilized SGC-7901 and MGC-803 gastric cancer cell lines as research models and constructed stable cell lines overexpressing ATF3. We conducted a series of assays including cell proliferation, colony formation, cell migration, tumorsphere formation, and invasion to investigate the functional roles of ATF3 in stemness of gastric cancer. The possible effect of ATF3 on epithelial-mesenchymal transition (EMT) was assessed through flow cytometry and qRT-PCR. In vivo functional effect of upregulation of ATF3 on tumor growth was examined in a mouse xenograft model. Results We found that overexpression of ATF3 inhibited cell proliferation, colony formation, cell migration and invasion. In addition, up-regulation of ATF3 attenuated tumorsphere formation, cell stemness, and potentially decreased expression of EMT markers. Moreover, ATF3 overexpression inhibited tumorigenesis in mouse xenograft model. Conclusion Our data suggest a suppressive role of ATF3 in gastric cancer development. Our findings will provide a potential therapeutic strategy and novel drug target for gastric cancer.