关键词： apoptosis   ATF   cisplatin   ERK1   Noxa   HEPATOCELLULAR-CARCINOMA   INDUCED APOPTOSIS   CANCER   TRANSCRIPTION   DEATH   THERAPY   FAMILY   NEUROBLASTOMA   FLUOROURACIL   CARBOPLATIN  

摘要： The platinum-based DNA damaging agent cisplatin is used as a standard therapy for locally advanced head and neck squamous cell carcinoma (HNSCC). However, the mechanisms underpinning the cytotoxic effects of this compound are not entirely elucidated. Cisplatin produces anticancer effects primarily via activation of the DNA damage response, followed by inducing BCL-2 family dependent mitochondrial apoptosis. We have previously demonstrated that cisplatin induces the expression of proapoptotic BCL-2 family protein, Noxa, that can bind to the prosurvival BCL-2 family protein, MCL-1, to inactivate its function and induce cell death. Here, we show that the upregulation of Noxa is critical for cisplatin-induced apoptosis in p53-null HNSCC cells. This induction is regulated at the transcriptional level. With a series of Noxa promoter-luciferase reporter assays, we find that the CRE (cAMP response element) in the promoter is critical for the Noxa induction by cisplatin treatment. Among the CREB/ATF transcription factors, ATF3 and ATF4 are induced by cisplatin, and downregulation of ATF3 or ATF4 reduced cisplatin-induced Noxa. ATF3 and ATF4 bind to and cooperatively activate the Noxa promoter. Furthermore, ERK1 is involved in cisplatin-induced ATF4 and Noxa induction. In conclusion, ATF3 and ATF4 are important regulators that induce Noxa by cisplatin treatment in a p53-independent manner.

关键词： ATF3   TNF-alpha   Osteoblast   Differentiation   Inflammation   NECROSIS-FACTOR-ALPHA   NF-KAPPA-B   TRANSCRIPTIONAL REGULATION   SKELETAL DEVELOPMENT   RUNX2   EXPRESSION   CELLS  

摘要： Tumor necrosis factor (TNF)-alpha, which is a proinflammatory cytokine, inhibits osteoblast differentiation under diverse inflammatory conditions. Activating transcription factor 3 (ATF3), which is a member of the ATF/cAMP response element-binding protein family of transcription factors, has been implicated in the regulation of cell proliferation and differentiation. However, the precise interactions between ATF3 and the TNF-alpha signaling pathway in the regulation of osteoblast differentiation remain unclear. In this study, we examined the role of ATF3 in the TNF-alpha-mediated inhibition of osteoblast differentiation and investigated the signaling pathways involved. The treatment of cells with TNF-alpha downregulated osteogenic markers, but significantly upregulated the expression of Atf3. The inhibition of Atf3 by small interfering RNAs rescued osteogenesis, which was inhibited by TNF-alpha. Conversely, the enforced expression of Atf3 enhanced the TNF-alpha-mediated inhibition of osteoblast differentiation, as revealed by the measurement of osteogenic markers and alkaline phosphatase staining. Mechanistically, TNF-alpha-induced Atf3 expression was significantly suppressed by the inhibition of the c-Jun N-terminal kinase (JNK) pathway. Furthermore, the overexpression of Atf3 did not affect the rescue effect that inhibiting TNF-alpha expression using a JNK inhibitor had on alkaline phosphatase activity and mineralization. Taken together, these results indicate that ATF3 mediates the inhibitory action of TNF-alpha on osteoblast differentiation and that the TNF-alpha-activated JNK pathway is responsible for the induction of Atf3 expression. (C) 2018 Elsevier Inc. All rights reserved.

关键词： Breast cancer   NRP-1   ABCG2   TNC   Integrin beta 3   Chemoresistance   Adriamycin   Cyclophosphamide   EPITHELIAL-MESENCHYMAL TRANSITION   NF-KAPPA-B   ACTIVATION   SUPPRESSES   EXPRESSION   ADHESION   GROWTH   ATF3   AKT   PHOSPHORYLATION  

摘要： Background: Neuropilin-1 (NRP-1), a non-tyrosine kinase glycoprotein receptor, is associated with poor prognosis breast cancer, however transcriptomic changes triggered by NRP-1 overexpression and its association with chemoresistance in breast cancer have not yet been explored. Methods: BT-474 NRP-1 variant cells were generated by stable overexpression of NRP-1 in the BT-474 breast cancer cell line. RNA sequencing and qRT-PCR were conducted to identify differentially expressed genes. The role of an upregulated oncogene, Tenascin C (TNC) and its associated pathway was investigated by siRNA-mediated knockdown. Resistant variants of the control and BT-474 NRP-1 cells were generated by sequential treatment with four cycles of Adriamycin/Cyclophosphamide (4xAC) followed by four cycles of Paclitaxel (4xAC + 4xPAC). Results: NRP-1 overexpression increased cellular tumorigenic behavior. RNA sequencing identified upregulation of an oncogene, Tenascin-C (TNC) and downregulation of several tumor suppressors in BT-474 NRP-1 cells. Additionally, protein analysis indicated activation of the TNC-associated integrin beta 3 (ITGB3) pathway via focal adhesion kinase (FAK), Akt (Ser473) and nuclear factor kappa B (NF-kB) p65. siRNA-mediated TNC knockdown ablated the migratory capacity of BT-474 NRP-1 cells and inactivated FAK/Akt473 signaling. NRP-1 overexpressing cells downregulated breast cancer resistance protein (BCRP/ABCG2). Consequently, sequential treatment with Adriamycin/Cyclophosphamide (AC) cytotoxic drugs to generate resistant cells indicated that BT-474 NRP-1 cells increased sensitivity to treatment by inactivating NRP-1/ITGB3/FAK/Akt/NF-kB p65 signaling compared to wild-type BT-474 resistant cells. Conclusions: We thus report a novel mechanism correlating high baseline NRP-1 with upregulated TNC/ITGB3 signaling, but decreased ABCG2 expression, which sensitizes BT-474 NRP-1 cells to Adriamycin/Cyclophosphamide. The study emphasizes on the targetability of the NRP-1/ITGB3

关键词： ATF3   Particulate matter   Inflammation   Airway epithelial cells   NF-kappa B   AP-1   ACTIVATING TRANSCRIPTION FACTOR-3   NF-KAPPA-B   PULMONARY INFLAMMATION   NEGATIVE REGULATOR   SIGNALING PATHWAYS   EPITHELIAL-CELLS   EXPRESSION   NANOPARTICLES   PARTICLES   EXPOSURE  

摘要： Airborne particulate matter (PM) has been reported to be associated with a wide range of respiratory disorders. However, the mechanisms underlying PM-induced airway inflammation remain largely unknown. Generally, ATF3 negatively regulates pro-inflammatory cytokines production in response to TLR4 signaling. Here we first showed ATF3 has promoting effects in PM-induced airway inflammation in vitro an in vivo. We demonstrated PM significantly upregulated ATF3 expression in HBE cells and in mouse lung tissues. ATF3 siRNA markedly inhibited, while ATF3-recombinant over-expression plasmid significantly increased PM-induced IL-6 expression in cultured HBE cells, and PM-induced IL-6, CXCL2 expression as well as neutrophil infiltration, mucus overproduction in the lung of ATF3(-/-) mice were all notably reduced relative to the wild-type littermates. Furthermore, we showed ATF3 mediated PM-induced inflammatory cytokines expression partly through NF-kappa B and AP-1 pathways. Our data further elucidates the mechanisms underlying PM-induced airway inflammation, and indicates ATF3 may function as different role in response to different stimuli.

关键词： 成纤维细胞   ATF3   心脏   小鼠   条件性敲除  

摘要： 目的:构建成纤维细胞特异敲除转录因子ATF3基因的小鼠模型。方法:利用胚胎显微注射法构建在ATF3基因携带loxP位点的小鼠(ATF3^(fl/fl)),以成纤维细胞细胞特异性表达的Col1a2-Cre介导ATF3条件性敲除,筛选出成纤维细胞条件性敲除ATF3的小鼠(Col1a2-ERCre:ATF3^(fl/fl));取鼠尾DNA进行PCR鉴定基因型,腹腔注射他莫昔芬诱导该基因敲除;12只敲除小鼠和12只同窝野生型小鼠各分为心脏缺血再灌注(I/R)组和假手术(sham)组,术后2h取心脏组织研磨后流式细胞分选成纤维细胞,进而提取mRNA并反转为cDNA,采用实时荧光定量PCR检测ATF3的表达;另取4只敲除小鼠和4只同窝野生型小鼠复制I/R模型,术后6h取心脏组织进行免疫荧光染色,观察成纤维细胞ATF3表达。结果:鼠尾DNA PCR鉴定基因型结果显示Cre基因型为阳性,ATF3 loxP基因型为纯合阳性;与假手术组相比,野生型小鼠术后心脏成纤维细胞ATF3表达明显升高[(0.02534±0.002654)vs.(0.5982±0.03495),P<0.0001],与术后野生型小鼠相比,术后条件性敲除组心脏成纤维细胞ATF3表达明显降低[(0.5982±0.03495)vs.(0.06456±0.005704),P<0.0001];免疫荧光共定位染色显示野生型小鼠心脏成纤维细胞ATF3蛋白表达,条件性敲除小鼠ATF3蛋白未见表达。结论:成纤维细胞特异敲除ATF3小鼠构建成功,为进一步实验奠定基础。

关键词： Joint inflammatory pain   DRG neurons   ATF3   P2Xreceptors   SGC activation   HSP90 inhibition   DORSAL-ROOT GANGLION   ACTIVATING TRANSCRIPTION FACTOR-3   SATELLITE GLIAL-CELLS   P2X3 RECEPTORS   INFLAMMATORY RESPONSES   SIGNALING PATHWAYS   P2X(7) RECEPTOR   SENSORY NEURONS   MESSENGER-RNA   STRESS  

摘要： Purinergic receptors (P2XRs) have been widely associated with pain states mostly due to their involvement in neuron-glia communication. Interestingly, we have previously shown that satellite glial cells (SGC), surrounding dorsal root ganglia (DRG) neurons, become activated and proliferate during monoarthritis (MA) in the rat. Here, we demonstrate that P2X7R expression increases in ipsilateral DRG after 1 week of disease, while P2X3R immunoreactivity decreases. We have also reported a significant induction of the activating transcriptional factor 3 (ATF3) in MA. In this study, we show that ATF3 knocked down in DRG cell cultures does not affect the expression of P2X7R, P2X3R, or glial fibrillary acidic protein (GFAP). We suggest that P2X7R negatively regulates P2X3R, which, however, is unlikely mediated by ATF3. Interestingly, we found that ATF3 knockdown in vitro induced significant decreases in the heat shock protein 90 (HSP90) expression. Thus, we evaluated in vivo the involvement of HSP90 in MA and demonstrated that the HSP90 messenger RNA levels increase in ipsilateral DRG of inflamed animals. We also show that HSP90 is mostly found in a cleaved form in this condition. Moreover, administration of a HSP90 inhibitor, 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG), attenuated MA-induced mechanical allodynia in the first hours. The drug also reversed the HSP90 upregulation and cleavage. 17-DMAG seemed to attenuate glial activation and neuronal sensitization (as inferred by downregulation of GFAP and P2X3R in ipsilateral DRG) which might correlate with the observed pain alleviation. Our data indicate a role of HSP90 in MA pathophysiology, but further investigation is necessary to clarify the underlying mechanisms.

关键词： Lidocaine   Sufentanil   Activating transcription factor 3 (ATF3)   Neurotoxicity   ROPIVACAINE CAUSES NEUROTOXICITY   HYPERALGESIA   BUPIVACAINE   ATF3   CORD   TRANSCRIPTION   INDUCTION   TOLERANCE   ADJUVANTS   ROLES  

摘要： Spinal anesthesia has evolved into a safe and widely accepted method of anesthesia. Synergy between opioids and local anesthetics further increases the quality of analgesia and decreases the dose requirement of both local anesthetics and opioids. However, over the last decades compelling evidence suggested that lidocaine could be more neurotoxic than other commonly used local anesthetics. Whether opioids can modify the local anesthetics-induced neurotoxicity is largely unexplored. Here, we investigated the effect of sufentanil on the neurotoxicity induced by intrathecal lidocaine in a rat model. Our data showed that 5 A mu g/ml sufentanil didn't deteriorate nor reduce the histopathological injuries induced by intrathecal application of 10% lidocaine in a rat model. However, it did alleviate sensory and motor function impairments induced by 10% lidocaine. Repeated intrathecal injection of 5 A mu g/ml sufentanil also decreased the paw withdraw threshold compared to the baseline. An increase in expression of activating transcription factor 3, a stress response gene, as a marker for injured neurons, was also detected in lidocaine-induced neurotoxicity, while 5 A mu g/ml sufentanil inhibited lidocaine-induced the upregulation of activating transcription factor 3. These results suggest that sufentanil alleviates lidocaine induced sensory and motor impairments, and did not worsen histopathological injury induced by intrathecal lidocaine.

关键词： KAPPA-B-ZETA   SUCCINATE-DEHYDROGENASE   OXIDATIVE STRESS   KEY REGULATOR   PSORIASIS   OXYGENASE-1   EXPRESSION   APOPTOSIS   CELLS   HEME  

摘要： Metabolic regulation has been recognized as a powerful principle guiding immune responses. Inflammatory macrophages undergo extensive metabolic rewiring(1) marked by the production of substantial amounts of itaconate, which has recently been described as an immunoregulatory metabolite(2). Itaconate and its membrane-permeable derivative dimethyl itaconate (DI) selectively inhibit a subset of cytokines(2), including IL-6 and IL-12 but not TNF. The major effects of itaconate on cellular metabolism during macrophage activation have been attributed to the inhibition of succinate dehydrogenase(2,3), yet this inhibition alone is not sufficient to account for the pronounced immunoregulatory effects observed in the case of DI. Furthermore, the regulatory pathway responsible for such selective effects of itaconate and DI on the inflammatory program has not been defined. Here we show that itaconate and DI induce electrophilic stress, react with glutathione and subsequently induce both Nrf2 (also known as NFE2L2)-dependent and -independent responses. We find that electrophilic stress can selectively regulate secondary, but not primary, transcriptional responses to toll-like receptor stimulation via inhibition of I kappa B zeta protein induction. The regulation of I kappa B zeta is independent of Nrf2, and we identify ATF3 as its key mediator. The inhibitory effect is conserved across species and cell types, and the in vivo administration of DI can ameliorate IL-17-I kappa B zeta driven skin pathology in a mouse model of psoriasis, highlighting the therapeutic potential of this regulatory pathway. Our results demonstrate that targeting the DI-I kappa B zeta regulatory axis could be an important new strategy for the treatment of IL-17-I kappa B zeta-mediated autoimmune diseases.

关键词： JUN DIMERIZATION PROTEIN-2