关键词： Bladder cancer   Cytoskeleton reorganization   TNF-alpha   METTL3   CLASP2   IQGAP1  

摘要： Background: Bladder cancer (BCa) metastasis is a multi-step process triggered by cytoskeleton reorganization. However, the regulation mechanism of cytoskeleton reorganization in BCa remains ambiguous. This study elucidated the influence of tumor necrosis factor-alpha (TNF-alpha) in cytoskeleton remodeling during BCa metastasis and its possible mechanisms. Methods: Colony formation, scratch, transwell, and the nude mouse model were adopted to evaluate the growth and metastasis. Molecular expression was assessed by immunohistochemical staining, quantitative real-time PCR (qRT-PCR), and Western blotting. The N6-methyladenosine (m6A) level was detected by methylated RNA immunoprecipitation (MeRIP). Protein interaction was validated by Co-immunoprecipitation (Co-IP). Immunofluorescence staining was used to identify rearrangement of actin filament fibers (F-actin) and protein colocalization. Results: TNF-alpha facilitated cytoplasmic linker associated protein 2 (CLASP2) and methyltransferase like 3 (METTL3) expression in a dose (10-50 ng/mL)-dependent manner in BCa. CLASP2 high expression suggested a shorter overall survival of BCa patients. CLASP2 deficiency suppressed BCa cell proliferation, migration, and invasion via disrupting F-actin cytoskeleton. Mechanistically, TNF-alpha promoted METTL3-mediated m6A modification of CLASP2 to enhance CLASP2 mRNA stability. Moreover, CLASP2 directly interplayed with IQ motif containing GTPase activating protein 1 (IQGAP1) to regulate F-actin cytoskeleton remodeling. In vivo data demonstrated that inhibition of METTL3/CLASP2 axis delayed lung metastasis in nude mice. Conclusion: TNF-alpha favors BCa cell metastasis, which involves METTL3-mediated m6A modification of CLASP2 that interacts with IQGAP1, thus leading to F-actin cytoskeleton remodeling. Our findings suggest targeting TNF alpha/METTL3/CLASP2/IQGAP1 axis as a potential avenue for promising treatment for BCa.

摘要： This study employed a multi-omics integration approach to identify circulating biomarkers for gastric cancer (GC). We analyzed plasma and tumor tissue single-cell RNA sequencing data, along with gene and protein quantitative trait loci analyses. Leveraging data from UK Biobank and FinnGen, we investigated genetic associations with GC. Through colocalization, Mendelian Randomization, and various filtering analyses, we identified four genes (IQGAP1, KRTCAP2, PARP1, MLF2) and four proteins (EGFL9 [DLK2], ECM1, PDIA5, TIMP4) as potential GC biomarkers. These were selected based on significant genetic colocation probabilities and significant associations with GC. Seven of these biomarkers demonstrated predictive capability for GC occurrence, with AUC ranging from 0.61 to 0.99. Drug prediction analysis identified seven protein biomarkers as potential targets for immunotherapy, targeted therapies, and tumor chemotherapy. Further scRNA-seq analysis revealed significant expression differences between gastric tumor and normal tissues, particularly the upregulation of IQGAP1, which highlights its role in tumor growth.

关键词： Hippo pathway   IQGAP1   malignant melanoma   Pannexin 1   YAP  

摘要： In this study, we explored the intricate relationship between Pannexin 1 (PANX1) and the Hippo signaling pathway effector, Yes-associated protein (YAP). Analysis of The Cancer Genome Atlas (TCGA) data revealed a significant positive correlation between PANX1 mRNA and core Hippo components, Yes-associated protein 1 [YAP], Transcriptional coactivator with PDZ-binding motif [TAZ], and Hippo scaffold, Ras GTPase-activating-like protein IQGAP1 [IQGAP1], in invasive cutaneous melanoma and breast carcinoma. Furthermore, we demonstrated that PANX1 expression is upregulated in invasive melanoma cell lines and is associated with increased YAP protein levels. Notably, our investigations uncovered a previously unrecognized interaction between endogenous PANX1 and the Hippo scaffold protein IQGAP1 in melanoma cells. Moreover, our findings revealed that IQGAP1 exhibits differential expression in melanoma cells and plays a regulatory role in cellular morphology. Functional studies involving PANX1 knockdown provided compelling evidence that PANX1 modulates YAP protein levels and its cotranscriptional activity in melanoma and breast carcinoma cells. Importantly, our study highlights the potential therapeutic significance of targeting PANX1. Pharmacological inhibition of PANX1 using selective FDA-approved inhibitors or PANX1 knockdown reduced YAP levels in melanoma cells. Furthermore, our Clariom (TM) S analysis unveiled key genes implicated in cell proliferation, such as neuroglin1 (NRG1), beta-galactoside binding protein and galectin-3 (LGALS3), that are affected in PANX1-deficient cells. In summary, our investigation delves into the intricate interplay between PANX1 and YAP in the context of invasive melanoma, offering valuable insights into potential therapeutic strategies for effective treatment.

关键词： CDC42   IQGAP1   Phagocytosis   Apostichopus japonicus   Innate immunity   Invertebrate   Aquaculture health  

摘要： Cell Division Cycle 42 protein (CDC42) is a pivotal member of the Rho family of small GTPases, integral to cell signaling and the regulation of cell polarity. CDC42 is extensively documented to participate in host immunity, particularly in processes such as phagocytosis and cell migration. Nevertheless, the precise mechanisms by which CDC42 affects host-bacteria interactions remain unclear in Echinoderms. In this study, we have isolated and characterized the CDC42 gene from Apostichopus japonicus, referred to as AjCDC42. The complete cDNA sequence of AjCDC42 extends 1282 base pairs, including a typical classical RHO domain, spanning amino acids from 6 to 179. Spatial expression analysis demonstrated that AjCDC42 is ubiquitously expressed across all examined tissues. Following infection with Vibrio splendidus and stimulation with LPS, the protein and mRNA levels of AjCDC42 were significantly upregulated in coelomocytes. Moreover, the knockdown of AjCDC42 significantly decreased V. splendidus-induced phagocytic activity in coelomocytes, leading to a reduced intracellular load of V. splendidus, as evidenced by flow cytometry and bacterial plate counting assays. Mechanistically, AjCDC42 was identified as an interacting partner of IQ Motif Containing GTPase Activating Protein 1 (AjIQGAP1) through the use of co-immunoprecipitation, immunofluorescence, and GST pull-down assays combined with mass spectrometry techniques. Additional functional analyses revealed that the knockdown of AjIQGAP1 similarly decreased the phagocytic activity and the intracellular load of V. splendidus in coelomocytes. Furthermore, we detected that decreased levels of AjCDC42 and AjIQGAP1 impaired cytoskeletal rearrangement and delayed lysosomal clearance of V. splendidus. In summary, our findings underscore the pivotal role of the AjCDC42AjIQGAP1 axis in regulating coelomocytes phagocytosis in A. japonicus during V. splendidus infection.

关键词： IQGAP1   neutrophils   CD11c   integrin  

摘要： IQ motif containing GTPase activating protein 1 (IQGAP1) is a protein scaffold that integrates signals regulating various cellular functions. Recently, utilizing proteomics as a discovery tool and co-immunoprecipitation as a validation method, we reported IQGAP1 as a potential ligand for CD11c, an adhesion molecule that was highly expressed in the intracellular components of neutrophils and regulated the maturation, survival, and function of neutrophils. To date, the role of IQGAP1 in inflammation and immune response is largely unknown. In this study, we reported the direct binding of CD11c with IQGAP1 through various methods. In addition, we investigated the role of IQGAP1 in neutrophil functions by using IQGAP1 knockout (KO) mice. We discovered that IQGAP1 deficiency led to impaired bone marrow neutrophil maturation, reactive oxygen species (ROS) generation, phagocytosis, and neutrophil extracellular traps formation. Thus, for the first time, we reported the essential role of IQGAP1 in neutrophil development.

关键词： Ischemic stroke   Platelet-related genes   IQGAP1   Diagnostic marker   Immune cells  

摘要： Introduction: Ischemic stroke (IS) is a complex illness resulting from a combination of numerous environmental and genetic risk factors. Recent reports have shed light on the vital role that platelets play in the pathophysiology of IS. Here, we aimed to explore the potential platelet-related genes in IS and investigate the effect of plateletrelated genes in the immune microenvironment of IS. Methods: The data of IS were retrieved from the Gene Expression Omnibus database. Firstly, we screened the platelet-related genes that were correlated with the onset of IS using differential expression analysis, enrichment analyses, and protein-protein interaction (PPI) network. Moreover, we analyzed the clinical value and functional information of platelet-related genes in IS. Finally, we explored the correlation between platelet-related genes and immune cells' infiltration. Results: Ten platelet-related genes that were correlated with the onset of IS were identified, among which IQGAP1 was located at the core of the PPI network. IQGAP1 was found to be expressed in the normal brain tissue and its expression was significantly elevated in IS samples. The area under the curve (AUC) values for IQGAP1 in both the GSE16561 and GSE58294 datasets were close to 1. IQGAP1 knockdown might increase OGD/R-induced HT22 cell viability. Additionally, FoxO signaling pathway, NOD-like receptor signaling pathway, Phagosome and Platelet activation pathways were significantly activated in IS patients with high IQGAP1 expression compared to those with low IQGAP1 expression. The IS patients in the IQGAP1high and IQGAP1low groups showed dramatically different proportions of immune cells and immune-related functions, and the IQGAP1 expression was correlated with the immune cell' infiltration in IS. Conclusions: In this study, we identified the IQGAP1 might serve as a potential diagnostic marker for IS, and the IQGAP1 expression was very relevant in determining the immune cell' infiltration in IS patien

关键词： 宫颈癌   半胱氨酸天冬氨酸蛋白酶1   IQ序列三磷酸鸟苷酶激活蛋白   血管内皮生长因子C  

摘要： 目的探究半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、IQ序列三磷酸鸟苷酶激活蛋白(IQGAP1)及血管内皮生长因子C(VEGF-C)在宫颈癌中的表达对患者预后的预测价值。方法选取2021年6月—2022年6月于丽水市人民医院进行宫颈癌治疗的患者180例,经手术提取宫颈癌组织作为实验组,同时搜集距离癌组织3 cm处的癌旁组织作为对照组。采用免疫组化法测定Caspase-1、IQGAP1及VEGF-C阳性表达情况,并分析其与临床特征和预后之间的关系。结果癌变组织Caspase-1、IQGAP1及VEGF-C阳性表达率(66.67%、72.22%及61.11%)与癌旁组织(39.44%、42.22%及36.11%)比较均升高,差异均有统计学意义(χ^(2)=26.778、33.090及22.517,均P<0.05)。宫颈癌癌变组织中Caspase-1、IQGAP1及VEGF-C表达与分化程度、临床分期、淋巴结转移相关(均P<0.05)。将宫颈癌患者年龄、体质量指数、瘤体直径及组织类型等因素进行控制后,Caspase-1、IQGAP1及VEGF-C仍与分化程度、临床分期、淋巴结转移显著相关(均P<0.05)。受试者特征曲线(ROC)分析Caspase-1、IQGAP1、VEGF-C及联合检测对宫颈癌预后的预测价值,其曲下面积分别为0.707、0.752、0.795及0.911。结论Caspase-1、IQGAP1及VEGF-C在宫颈癌癌变组织中均呈现上调状态,并且其表达高低变化与分化程度、临床分期、淋巴结转移和预后情况之间具有一定联系,可能共同参与该疾病的发生、发展,并对患者的预后情况产生影响。

关键词： Cancer-predisposing variant   Cingulin   Epithelial-mesenchymal transition   IQGAP1   Rac1  

摘要： Background: Cingulin (CGN) is a pivotal cytoskeletal adaptor protein located at tight junctions. This study investigates the link between CGN mutation and increased cancer susceptibility through genetic and mechanistic analyses and proposes a potential targeted therapeutic approach. Methods: In a high-cancer-density family without known pathogenic variants, we performed tumor-targeted and germline whole-genome sequencing to identify novel cancer-associated variants. Subsequently, these variants were validated in a 222 cancer patient cohort, and CGN c.3560C > T was identified as a potential cancer-risk allele. Both wild-type (WT) (c.3560C > C) and variant (c.3560C > T) were transfected into cancer cell lines and incorporated into orthotopic xenograft mice model for evaluating their effects on cancer progression. Western blot, immunofluorescence analysis, migration and invasion assays, two-dimensional gel electrophoresis with mass spectrometry, immunoprecipitation assays, and siRNA applications were used to explore the biological consequence of CGN c.3560C > T. Results: In cancer cell lines and orthotopic animal models, CGN c.3560C > T enhanced tumor progression with reduced sensitivity to oxaliplatin compared to the CGN WT. The variant induced downregulation of epithelial marker, upregulation of mesenchymal marker and transcription factor, which converged to initiate epithelial-mesenchymal transition (EMT). Proteomic analysis was conducted to investigate the elements driving EMT in CGN c.3560C > T. This exploration unveiled overexpression of IQGAP1 induced by the variant, contrasting the levels observed in CGN WT. Immunoprecipitation assay confirmed a direct interaction between CGN and IQGAP1. IQGAP1 functions as a regulator of multiple GTPases, particularly the Rho family. This overexpressed IQGAP1 was consistently associated with the activation of Rac1, as evidenced by the analysis of the cancer cell line and clinical sample harboring CGN c.3560C > T. Notably, acti

关键词： Phosphate glasses   Glass-ceramic   Solid electrolyte   Crystallization kinetic   Nasicon structure   Impedance spectroscopy  

摘要： Advancing energy density, enabling lithium metal anodes, and ensuring unparalleled safety and operational reliability in lithium batteries hinge on advancing inorganic solid-state electrolytes. To overcome current impediments, we present an innovative approach that integrates glass-ceramics with a pioneering new Nasicon strategy involving molybdenum doping. In the conducted study, a series of 14Li2O-9Al2O3-38TiO2-(39-x)P2O5xMoO3 glasses, denoted as LATPMox, along with their corresponding glass-ceramics (LATPMox-GC), have exhibited a promising characteristic as solid electrolytes. X-ray diffraction (XRD) analysis confirms the formation of the novel Mo-doped Nasicon phases in the glass-ceramics, as validated by Rietveld refinement. Examination of the crystallization kinetic behavior of the glasses reveals a three-dimensional nucleation process with spherical particle growth, featuring an activation energy of 165 kJ mol-1. Transmission Electron Microscopy TEM characterization aligns crystallization behavior with crystallite and distribution within the glass matrix, resulting in a compact and dense microstructure. The structural properties of the resultant phases are examined through FT-IR, Raman spectroscopy, and TEM-SEAD analysis. Vickers indentation tests were employed to assess the microscopic fracture toughness, and both the glass and glass-ceramics materials demonstrated favorable mechanical performance. Optical characterization using UV-visible absorption highlights the reduction of Mo6+ to Mo5+, likely occupying tetrahedral sites within the crystalline lattice. Impedance spectroscopy measurement showcases the effective promotion of ionic conductivity following Mo doping, reaching a total conductivity value of 5.50 x 10-5 Omega- 1 cm- 1 along with a high lithium transference number of 0.99 at room temperature for LATPMo2.6-GC glass-ceramic. This value is larger than that of many other glass-ceramics as well as that of the well-known lithium phosphorous oxy-nitrid

关键词： IQGAP1   B cell   MAPK   IQGAP1 signalosome  

摘要： IQGAP1 is a multi-functional scaffold protein. However, its role in B cell development and function is unknown. Here, we show IQGAP1 as an essential scaffold that regulates early B cell development and function. Iqgap1-/- mice contained significantly increased numbers of B220+ B, B220+IgM- pro/pre-B, and B220LowIgM+ immature-B cells in the bone marrow. In the spleens of the Iqgap1-/- mice, newly formed and follicular B cell numbers were increased, while the marginal zone B cell numbers were significantly reduced. Lack of IQGAP1 reduced T-dependent and T-independent humoral responses. Mechanistically, the lack of IQGAP1 considerably decreased the phosphorylation of Mek1/2, Erk1/2, and Jnk1/2. B cells from Iqgap1-/- mice failed to suppress IL-7R-mediated activation of Stat5a/b, an essential step for cell-cycle exit and initiate light-chain recombination, reducing RS rearrangement frequency. Our study provides the first evidence that IQGAP1-based signalosome is necessary for the development and functions of B cells.