关键词： ATF3   HPV   MMP1   SHARP1   ROS   DSBs   Cervical cancer  

摘要： BackgroundActivating Transcription Factor 3 (ATF3) is known for its tumor-suppressive properties in cervical cancer, particularly through its role in stress response and interactions with human papillomavirus (HPV) oncogenes. This study investigates ATF3's regulatory impact on metastasis-related genes, oxidative stress, and DNA damage in HPV-positive cervical cancer cells. MethodsHeLa and Ca Ski cell lines were transfected with ATF3-expressing vectors. Western blotting and quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to confirm ATF3 overexpression following transfection. ROS assays and Comet assays assessed the impact of ATF3 on oxidative stress and DNA damage, while RT-qPCR was used to evaluate changes in HPV E6/E7, SHARP1, and MMP1 gene expression. ResultsATF3 overexpression led to elevated ROS levels (p < 0.02), resulting in oxidative DNA damage. These results demonstrate ATF3's cytotoxic impact on cervical cancer cells through oxidative stress and DNA damage. Additionally, ATF3 overexpression significantly decreased MMP1 expression (p < 0.03), indicating a potential anti-metastatic effect, while SHARP1 and HPV E6/E7 expression levels were not significantly altered, indicating selective gene modulation by ATF3. ConclusionThese findings reveal that ATF3 contributes to tumor suppression in cervical cancer by modulating oxidative stress and DNA damage, selectively targeting genes involved in metastasis. These findings supports ATF3's role in regulating key pathways in HPV-positive cervical cancer cells, providing a basis for further exploration of ATF3 as a target in therapeutic strategies aimed at improving outcomes in cervical cancer.

关键词： Common clinical tumors   Targeted drug screening   Personalized medicine   Patient-derived xenograft   PDX Model Database   PDX   patient-derived xenograft   ICH-GCP   International Council for Harmonization for Good Clinical Practice   NCI   National Cancer Institute   CDX   cell line-derived xenograft   CTC   scirculating tumor cells   BME   Basement Membrane Extract   EHS   Engelbreth-Holm-Swarm   LC   Lung cancer   NSCLCs   non-small cell lung cancers   LUAD   lung adenocarcinoma   LUSC   lung squamous cell carcinoma   SCLCs   small cell lung cancers   PI3K   phosphatidylinositol 3-kinase   AKT   serine/threonine kinase   HOX   homeobox   KRAS   kirsten rat sarcoma viral oncogene   HER3   human epidermal growth factor receptor 3   mTOR   mammalian target of rapamycin   ROR2   receptor tyrosine kinase-like orphan receptor 2   PM2.5   particulate matter 2.5   CDK   cyclin dependent kinase   ERK   extracellular regulated protein kinases   EGCG   epigallocatechin gallate   BRD4   bromodomain-containing protein 4   UBA1   ubiquitin-activating enzyme 1   MET   mesenchymal to epithelial transition factor   FGFR1   fibroblast growth factor receptor 1   MYCN   N-myc proto-oncogene   EGFR   epidermal growth factor receptor   CRC   Colorectal cancer   PVT1   plasmacytoma variant translocation 1   VEGFA   vascular endothelial growth factor A   TIMP2   tissue inhibitors of metalloproteinase 2   PDAP1   PDGFA-associated protein 1   PLOD2   Procollagen-Lysine 2-Oxoglutarate 5-Dioxygenase 2   SLC6A8   solute carrier family 6 member 8   MEK   mitogen-activated proteinkinase kinase   MDM2   murine double minute 2   PIM1   proviral integration site for Moloney murine leukemia virus 1   PROTAC   proteolysis targeting chimeric   HCC   Hepatocellular carcinoma   METTL5   methyltransferase 5, N6-adenosine   DANCR   differentiation antagonizing non-protein coding RNA   POSTN   Periostin   TGF-β1   transforming growth factor-β1   DPP4   dipeptidyl peptidase 4   DNAJB1   DnaJ heat shock protein family member B1   PRKACA   c-AMP dependant protein kinase A catalytic subunit α   YTHDF1   YTH N6-methyladenosine RNA binding protein 1   BCL-XL   B-cell lymphoma-XL   PHGDH   phosphoglycerate dehydrogenase   ROS1   C-ros oncogene 1   NMT1   N-myristoyl trans  

摘要： The establishment of mouse models is critical for discovering the biological targets of tumorigenesis and cancer development, preclinical trials of targeted drugs, and formulation of personalized therapeutic regimens. Currently, the patient-derived xenograft (PDX) model is considered a reliable animal tumor model because of its ability to retain the characteristics of the primary tumor at the histopathological, molecular, and genetic levels, and to preserve the tumor microenvironment. The application of the PDX model has promoted in-depth research on tumors in recent years, focusing on drug development, tumor target discovery, and precise treatment of patients. However, there are still some common questions. This review introduces the latest research progress and common questions regarding tumors with high mortality rates, focusing on their application in targeted drug screening and the formulation of personalized medical strategies. The challenges faced, improvement methods, and future development of the PDX model in tumor treatment applications are also discussed. This article provides technical guidance and comprehensive expectations for anti-cancer drug screening and clinical personalized therapy.

关键词： adipogenesis   ATF3   USP53   RhoA/ROCK pathway   transcriptional regulation  

摘要： Obesity is a widespread nutritional disorder, leading to a strong predisposition toward adverse health consequences. Activating transcription factor 3 (ATF3), a stress-induced transcription factor, has been documented as a therapeutic target for obesity. The intent of this project was to characterize the detailed role of ATF3 in adipogenesis in the context of obesity and its obscure downstream mechanism. After adipogenic differentiation, RT-qPCR and western blot examined ATF3 and ubiquitin-specific peptidase 53 (USP53) mRNA levels and protein levels. Adipogenesis was identified by Oil Red O staining, triglyceride (TG) levels, and western blot analysis. JASPAR database, ChIP and luciferase reporter assays predicted and validated the transcriptional regulation of USP53 by ATF3. Western blot also examined the protein levels of Ras homolog family member A (RhoA)/Rho-associated coiled-coil kinase (ROCK) pathway-involved proteins. ATF3 mRNA and protein levels were depleted in the differentiated 3T3-L1 adipocytes, and ATF3 elevation hindered the adipogenesis of 3T3-L1 adipocytes. ATF3 suppressed the transcription of USP53 as a transcription factor and lowered USP53 expression. Eventually, USP53 upregulation partially blunted the inhibitory role of ATF3 overexpression in adipogenesis and the RhoA/ROCK pathway. Consequently, ATF3 might transcriptionally inactivate USP53 to repress adipocyte adipogenesis and downregulate the RhoA/ROCK pathway.

关键词： Gasoline exhaust particles   Activating transcription factor 3   Matrix metalloproteinase 1   Nicotinamide adenine dinucleotide phosphate oxidase 4   human umbilical vein endothelial cells   reactive oxygen species  

摘要： Gasoline exhaust particles (GEP) are risk factors for cardiovascular disease. Activating transcription factor 3 (ATF3) is a transcription factor known to form a heterodimer with AP-1 transcription factors for its target gene expression. However, the involvement of ATF3 in GEP-induced gene expression in human umbilical vein endothelial cells (HUVECs) has not been investigated. In this study, we found that GEP, at IC50 value of 59 mu g/ml, induced the expression of ATF3, which led to the expression of matrix metalloproteinase 1 (MMP1) in HUVECs. GEP induce an interaction between c-Jun and ATF3, and c-Jun depletion attenuates GEP-induced MMP1 expression. Depletion of NADPH oxidase 4 (Nox4) suppressed GEP-induced reactive oxygen species (ROS) generation and the subsequent upregulation of ATF3 and MMP1, suggesting that Nox4-derived ROS play a role as upstream regulators of GEP-induced ATF3 expression and MMP1 upregulation. Furthermore, Nox4 depletion attenuated the interaction between ATF3 and c-Jun and their binding to the AP-1 binding site of the MMP1 promoter. Taken together, these findings demonstrate that GEP induce the expression of MMP1 by generating Nox4-dependent ROS, which subsequently increase ATF3 expression and its interaction with c-Jun. This leads to their binding to the promoter region of MMP1 and its transcription. These findings suggest that Nox4-derived ROS and ATF3 are critical for GEP-induced MMP1 expression.

关键词： Myocardial ischemia-reperfusion injury   Melatonin   Ferroptosis   ATF3   GPX4  

摘要： Melatonin (MEL), functioning as a circulating hormone, is important for the regulation of ferroptosis in different health scenarios and acts as a crucial antioxidant in cardiovascular diseases. However, its specific function in ferroptosis related to myocardial ischemia-reperfusion injury (MIRI) remains to be fully elucidated. In our research, we utilized a rat model of MIRI induced by coronary artery ligation, along with a cell model subjected to hypoxia/reoxygenation (H/R). We evaluated relevant genes and proteins by real-time fluorescent quantitative PCR and Western blot analysis. To evaluate myocardial tissue damage and cell injury, we employed cell counting kit-8 assays, flow cytometry, hematoxylin-eosin staining, and 2,3,5-triphenyltetrazolium chloride staining techniques. Our results show that administering MEL notably reduces the concentrations of cTnT, CK-MB, and lactate dehydrogenase in the serum of MIRI rats, mitigates the extent of myocardial infarction, improves the recovery of pathological conditions in myocardial tissues, and reduces the concentrations of Fe2+, malondialdehyde (MDA), and reactive oxygen species (ROS) in the myocardial tissue, while also promoting increased glutathione levels. Moreover, MEL can also restore the reduced viability of H9C2 cells caused by H/R or ferroptosis inducers (RSL3), reduce the cellular content of Fe2+, MDA, and ROS, and inhibit ferroptosis. Mechanistically, MEL promotes the expression of GPX4 by downregulating the expression of ATF3, thereby inhibiting ferroptosis in cardiomyocytes and ultimately alleviating the process of MIRI. Our study demonstrates that MEL ameliorates MIRI by inhibiting ferroptosis.

关键词： Lupus nephritis   Ferroptosis   ATF3   Machine learning  

摘要： Background: The role of ferroptosis in lupus nephritis (LN) is unclear. This study aimed to explore the effects of ferroptosis-related genes in LN through bioinformatics prediction and experimental validation. Methods: Sample data were collected from the GEO dataset and divided into glomeruli and tubulointerstitium. We collected 382 ferroptosis-related genes. The intersection of ferroptosis-related genes with glomeruli and tubulointerstitium data, respectively, was performed. Machine learning methods (including unsupervised cluster typing and random forests) were operated to identify ferroptosis subtyping and ferroptosis important genes in LN. Immune infiltration and functional analysis were performed. The expression of ferroptosis important gene ATF3 was validated in vivo and in vitro. Results: 6 ferroptosis important genes common to glomeruli and tubulointerstitium were screened, including ATF3, CD44, CYBB, JUN, NCF2, and NNMT. ATF3 decreased in the LN group compared to the Control. Silencing ATF3 mitigated LPS/erastin-induced ferroptosis. Functional analysis showed that ATF3 was markedly enriched in the interferon-gamma-mediated signaling pathway, ECM-receptor interaction, and cell adhesion. In glomeruli, T cells regulatory (Tregs) infiltration decreased and Macrophages M1 levels increased with elevated ATF3 expression. Levels of immune cell infiltration were altered in different ferroptosis subtypes of LN glomeruli and tubulointerstitium. Conclusions: Ferroptosis-related ATF3 levels decreased in LN. Inhibition of ATF3 might alleviate LN development by affecting the macrophage M1 and Treg cell infiltration. These implied that ATF3 might be a potential target for developing LN therapeutic strategies.

关键词： 卵巢癌   绞股蓝皂苷L   mature-tRNA-Asp-GTC   pre-tRNA-Arg-TCT   铁死亡  

摘要： 目的：探讨mature-t RNA-Asp-GTC与pre-t RNA-Arg-TCT在卵巢癌（OC）细胞铁死亡表型中作用及绞股蓝皂苷L（Gyp-L）对OC细胞mature-t RNA-Asp-GTC与pre-t RNA-Arg-TCT的调控机制。方法：采用细胞增殖与活性检测法（CCK-8）检测人卵巢腺癌细胞（OVCAR3）增殖情况，计算顺铂(5、10、15、20、25、30μmol·L-1)、Gyp-L(25、50、75、100、125μmol·L-1)及在Gyp-L作用下顺铂的半数抑制浓度（IC50）以确定后续实验的干预浓度。细胞克隆实验与划痕实验反映OVCAR3细胞的增殖与迁移能力。PANDORA-seq小RNA测序检测Gyp-L干预后细胞差异表达的转运RNA衍生的小RNA（ts RNA），通过RNAhybrid软件查找ts RNA相应的靶基因。比色法或酶联免疫吸附测定法（ELISA）检测丙二醛（MDA）、谷胱甘肽（GSH）、脂质过氧化物（LPO）水平；Ferro Orange荧光探针检测Fe2+含量；DCFH-DA荧光探针检测活性氧（ROS）含量来共同反映OVCAR3细胞铁死亡的发生。将OVCAR3细胞分为空白组、50μmol·L-1Gyp-L组、100μmol·L-1Gyp-L组。实时荧光定量聚合酶链式反应（Real-timePCR）检测mature-t RNA-Asp-GTC、mature-t RNA-Leu-CAA、mature-mt＿t RNA-Tyr-GTA＿5＿end、mature-t RNAVal-CAC、mature-mt＿t RNA-Glu-TTC、pre-t RNA-Arg-TCT、mature-t RNA-Asn-GTT、羟甲基双烷合酶（HMBS）、Wnt、β-连环蛋白（β-catenin）、谷胱甘肽过氧化物酶4（GPX4）、Kelch样ECH关联蛋白1（KEAP1）、核转录因子红系2相关因子2（Nrf2）、活化转录因子3（ATF3）、胱氨酸/谷氨酸逆向转运蛋白（x CT）、溶血磷脂酰胆碱酰基转移酶（LPCAT3）、花生四烯酸15-脂氧合酶（ALOX15）基因表达；蛋白免疫印迹法（Western blot）检测HMBS、Wnt、β-catenin、GPX4、KEAP1、Nrf2、ATF3、x CT、LPCAT3、ALOX15蛋白表达。结果：50μmol·L-1Gyp-L组、100μmol·L-1Gyp-L组、顺铂组、50μmol·L-1Gyp-L+顺铂组及100μmol·L-1Gyp-L+顺铂组显著抑制OVCAR3细胞增殖与迁移（P<0.05）并加剧细胞铁死亡，发生体现为ROS、MDA、LPO、Fe2+含量增多及GSH含量明显降低（P<0.05）。与空白组比较，Gyp-L有效干扰OVCAR3细胞25种ts RNA的表达（P<0.05，|log2Fc|>1）;Gyp-L干预后，pre-t RNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4、pre-t RNA-Arg-TCT/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/LPCAT3/ALOX15轴表达均发生明显异常（P<0.05）。结论：pre-t RNA-Arg-TCT/HMBS/Wnt/β-catenin/GPX4、pre-t RNA-Arg-TCT/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、maturet RNA-Asp-GTC/LPCAT3/ALOX15信号通路参与OC发生发展。Gyp-L主要通过mature-t RNA-Asp-GTC/ATF3/KEAP1/Nrf2/x CT、mature-t RNA-Asp-GTC/LPCAT3/ALOX15信号轴激活OVCAR3细胞铁死亡发生，从而抑制OC发生发展。

关键词： Triple-negative breast cancer   Aurovertin B   Dual-specificity phosphatase 1   Activating transcription factor 3   Lung metastasis  

摘要： Triple-negative breast cancer (TNBC) is characterized by high mortality rates, primarily due to its propensity for metastasis. Addressing this challenge necessitates the development of effective antimetastatic therapies. This study aimed to identify natural compounds with potential antimetastatic properties mainly based on the high-throughput phenotypic screening system. This system, utilizing luciferase reporter gene assays combined with scratch wound assays, evaluates compounds based on their influence on the epithelialmesenchymal transition (EMT) marker E-cadherin. Through this approach, aurovertin B (AVB) was revealed to have significant antimetastatic capability. Notably, AVB exhibited substantial metastasis suppression in many TNBC cell lines, including MDA-MB-231, HCC1937, and 4T1. Also, its remarkable antimetastatic activity was demonstrated in vivo via the orthotopic breast cancer mouse model. Further exploration revealed a pronounced association between AVB-induced upregulation of dual-specificity phosphatase 1 (DUSP1) and its inhibitory effect on TNBC metastasis. Additionally, microarray analysis conducted to elucidate the underlying mechanism of the AVB-DUSP1 interaction identified activating transcription factor 3 (ATF3) as a critical transcription factor instrumental in DUSP1 transcriptional activation. This discovery, coupled with observations of enhanced ATF3-DUSP1 expression and consequent reduction in TNBC metastatic foci in response to AVB, provides novel insights into the metastatic mechanisms of TNBC. Significance Statement: This study constructs a high-throughput phenotypic screening system utilizing epithelial-mesenchymal transition marker E-cadherin promoter luciferase reporter gene combined with scratch wound assays. Aurovertin B was revealed to possess significant antimetastatic activity through this approach, which was further demonstrated via in vivo and in vitro experiments. The discovery of the regulatory role of the ATF3-DUSP1 pathway

关键词： aortic aneurysm and dissection   cGAS-STING pathway   DNA stability   micronuclei formation   P21  

摘要： BackgroundSporadic aortic aneurysm and dissection (AAD) is a critical condition characterised by the progressive loss of vascular smooth muscle cells (VSMCs) and the breakdown of the extracellular matrix. However, the molecular mechanisms responsible for the phenotypic switch and loss of VSMCs in AAD are not fully *** and resultsIn this study, we employed a discovery-driven, unbiased approach. This approach encourages us to explore the unknown functions of activating transcription factor 3 (ATF3) rather than merely confirming existing hypotheses, while no assumptions were made about ATF3 prior to the experiments. We ensured the unbiased nature of our assessment by conducting morphological evaluations with two independent observers in a blinded manner. We identified elevated expression of ATF3 in both human sporadic AAD tissues and mouse AAD models. VSMC-specific ATF3 conditional knockout (Atf3 cKO) mice showed notable enlargement, dissection and rupture in both thoracic and abdominal aortic regions after exposure to Ang II. Interestingly, older Atf3 cKO mice exhibited spontaneous aortic dissections and senescence of the aortic wall. Mechanistically, ATF3 deficiency led to the degradation of P21 through ubiquitination. Impaired DNA repair in VSMCs resulted in micronuclei formation in the cytoplasm, activating the cyclicGMP-AMP synthase- stimulator of interferon genes (cGAS-STING) pathway and inducing VSMC phenotypic switching and apoptosis. Finally, both pharmacological complementation of P21 function and knockdown of STING expression alleviated ATF3 deficiency-induced *** study indicates that ATF3 is essential for genomic DNA stability in VSMCs through the P21-cGAS-STING pathway, suggesting that enhancing ATF3 expression in VSMCs could help prevent sporadic *** points ATF3 deficiency led to degradation of P21 through ubiquitination, which abolished the G1 phase arrest. VSMCs had no time window to repair the damaged DNA, leading to g

关键词： Sjogren's syndrome   Apigenin   Salivary gland epithelial cells   Ferroptosis  

摘要： Background: In Sjogren's ' s syndrome (SS)-an - an autoimmune disease characterized by dry mouth and eyes-salivary - salivary gland epithelial cells (SGECs) undergo ferroptosis, which disrupts their integrity and impairs saliva secretion. Apigenin, a phytoestrogen, is known to activate estrogen signalling and alleviate xerostomia in ovariectomized mice;however, its effect on SGEC survival and function in SS remains unclear. We hypothesized that apigenin alleviates SS symptoms and progression by inhibiting ferroptosis in SGECs and aimed to elucidate the underlying mechanism. Methods: Apigenin (50 mg/kg) was orally gavaged to non-obese diabetic (NOD)/LtJ female mice (SS model);changes in SS functional indicators were analyzed using mRNA sequencing and bioinformatic analyses of sub- mandibular glands. Interferon-gamma (IFN-gamma)-stimulated gamma )-stimulated SGECs were used to model SS in vitro;SGEC activity and aquaporin 5 (AQP5) expression were analyzed. Immunohistochemical staining, transmission electron microscopy, RT-qPCR, western blotting and other methods were used to verify the mechanisms. Results: Apigenin significantly increased salivary secretion and AQP5 expression while inhibiting ferroptosis and immune infiltration in NOD mouse submandibular glands. The oxidative stress gene ATF3 was upregulated and GPX4 was downregulated in NOD mice compared to that in control group (ICR mice);however, apigenin reversed this effect. IFN-gamma gamma treatment downregulated AQP5, SLC7A11, and GPX4 expression while promoting ATF3 expression and ferroptosis, which was mitigated by apigenin. ATF3 knockdown increased SLC7A11 and GPX4 expression, inhibiting SS and ferroptosis. Furthermore, apigenin inhibited ferroptosis in SGECs through ESR1 binding to ATF3. Conclusion: Apigenin alleviates SS by regulating SGEC ferroptosis via the ER alpha-regulated ATF3/SLC7A11 axis, highlighting its therapeutic potential in SS.