关键词： AKT   cervical cancer   IQGAP1   MMP16   nasopharyngeal cancer  

摘要： Human papillomavirus (HPV)-related cervical and nasopharyngeal cancers differ in molecular mechanisms underlying the oncogenic processes. The disparity may be attributed to differential expression of oncoproteins. The current study investigated the host oncogenes expression pattern in HPV-associated cervical and nasopharyngeal cancer. Formalin-fixed paraffin-embedded tissues originating from the nasopharyngeal and cervical regions were screened using Hematoxylin and Eosin staining. Genomic DNA and total RNA were extracted from confirmed cancer biopsies and non-cancer tissues (NC). HPV was detected by PCR using MY09/GP5+/6+ primers. Protein expression levels of AKT, IQGAP1, and MMP16 in HPV-infected cancers and controls were determined by immunohistochemistry. RT-qPCR was used to profile mRNAs of the oncogenes. AKT and IQGAP1 proteins were highly expressed in the epithelial cancers compared with the non-cancer tissues (p < 0.05). IQGAP1 and MMP16 mRNAs level was significantly higher in the cancers than in the NC (p < 0.05), but not AKT mRNA levels. MMP16 protein was ubiquitously expressed in all tissues. AKT mRNA level was significantly elevated in CC compared with NPC (p < 0.001). However, the difference in AKT, IQGAP1 and MMP16 proteins level between CC and NPC was not significant (p > 0.05). The oncoproteins expression level between the HPV-positive and HPV-negative cancer biopsies showed no significant difference (p < 0.05). Current study reports AKT but not IQGAP1 and MMP16 mRNAs differentially expression in cervical and nasopharyngeal cancers, independent of HPV infection status.

关键词： IQGAP1   Chemoresistance   Ferroptosis   YAP   Esophageal squamous cell carcinoma  

摘要： Chemoresistance is one of the major hindrances to many cancer therapies, including esophageal squamous cell carcinoma (ESCC). Ferroptosis, a new programmed cell death, plays an essential role in chemoresistance. IQdomain GTPase activating protein 1 (IQGAP1) is a scaffold protein and functions as an oncogene in various human malignancies. However, the underlying effect and molecular mechanisms of IQGAP1 on paclitaxel (PTX) resistance and ferroptosis in ESCC remain to be elucidated. In this study, we found that IQGAP1 was highly expressed in ESCC tissues and could as a potential biomarker for diagnosis and predicting the prognosis of ESCC. Functional studies revealed that IQGAP1 overexpression reduced the sensitivity of ESCC cells to PTX by enhancing ESCC cell viability and proliferation and inhibiting cell death, and protected ESCC cells from ferroptosis, whereas IQGAP1 knockdown exhibited contrary effects. Importantly, reductions of chemosensitivity and ferroptosis caused by IQGAP1 overexpression were reversed with ferroptosis inducer RSL3, while the increases of chemosensitivity and ferroptosis caused by IQGAP1 knockdown were reversed with ferroptosis inhibitor ferrostatin-1 (Fer-1) in ESCC cells, indicating that IQGAP1 played a key role in resistance to PTX through regulating ferroptosis. Mechanistically, we demonstrated that IQGAP1 overexpression upregulated the expression of Yes-associated protein (YAP), the central mediator of the Hippo pathway. YAP inhibitor Verteporfin (VP) could reverse the effects of IQGAP1 overexpression on ESCC chemoresistance and ferroptosis. Taken together, our findings suggest that IQGAP1 promotes chemoresistance by blocking ferroptosis through targeting YAP. IQGAP1 may be a novel therapeutic target for overcoming chemoresistance in ESCC.

关键词： Pancreatic cancer   Phosphoproteomics   Tumor typing   Tumor growth and metastasis   SOX4   MAPK1 kinase   IQGAP1 phosphorylation   KSEA analysis  

摘要： ObjectiveThis study aims to elucidate the functional role of IQGAP1 phosphorylation modification mediated by the SOX4/MAPK1 regulatory axis in developing pancreatic cancer through phosphoproteomics *** and phosphoproteomics data of pancreatic cancer were obtained from the Clinical Proteomic Tumor Analysis Consortium (CPTAC) database. Differential analysis, kinase-substrate enrichment analysis (KSEA), and independent prognosis analysis were performed on these datasets. Subtype analysis of pancreatic cancer patients was conducted based on the expression of prognostic-related proteins, and the prognosis of different subtypes was evaluated through prognosis analysis. Differential analysis of proteins in different subtypes was performed to identify differential proteins in the high-risk subtype. Clinical correlation analysis was conducted based on the expression of prognostic-related proteins, pancreatic cancer typing results, and clinical characteristics in the pancreatic cancer proteomics dataset. Functional pathway enrichment analysis was performed using GSEA/GO/KEGG, and most module proteins correlated with pancreatic cancer were selected using WGCNA analysis. In cell experiments, pancreatic cancer cells were grouped, and the expression levels of SOX4, MAPK1, and the phosphorylation level of IQGAP1 were detected by RT-qPCR and Western blot experiments. The effect of SOX4 on MAPK1 promoter transcriptional activity was assessed using a dual-luciferase assay, and the enrichment of SOX4 on the MAPK1 promoter was examined using a ChIP assay. The proliferation, migration, and invasion functions of grouped pancreatic cancer cells were assessed using CCK-8, colony formation, and Transwell assays. In animal experiments, the impact of SOX4 on tumor growth and metastasis through the regulation of MAPK1-IQGAP1 phosphorylation modification was studied by constructing subcutaneous and orthotopic pancreatic cancer xenograft models, as well as a liver metastas

摘要： Acute myeloid leukemia (AML) is fatal in the majority of adults. Identification of new therapeutic targets and their pharmacologic modulators are needed to improve outcomes. Previous studies had shown that immunization of rabbits with normal peripheral WBCs that had been incubated with fluorodinitrobenzene elicited high titer antibodies that bound to a spectrum of human leukemias. We report that proteomic analyses of immunoaffinity-purified lysates of primary AML cells showed enrichment of scaffolding protein IQGAP1. Immunohistochemistry and gene-expression analyses confirmed IQGAP1 mRNA overexpression in various cytogenetic subtypes of primary human AML compared to normal hematopoietic cells. shRNA knockdown of IQGAP1 blocked proliferation and clonogenicity of human leukemia cell-lines. To develop small molecules targeting IQGAP1 we performed in-silico screening of 212,966 compounds, selected 4 hits targeting the IQGAP1-GRD domain, and conducted SAR of the 'fittest hit' to identify UR778Br, a prototypical agent targeting IQGAP1. UR778Br inhibited proliferation, induced apoptosis, resulted in G2/M arrest, and inhibited colony formation by leukemia cell-lines and primary-AML while sparing normal marrow cells. UR778Br exhibited favorable ADME/T profiles and drug-likeness to treat AML. In summary, AML shows response to IQGAP1 inhibition, and UR778Br, identified through in-silico studies, selectively targeted AML cells while sparing normal marrow.

关键词： IQ结构域三磷酸鸟苷酶激活蛋白1   胰腺癌细胞   增殖   凋亡  

摘要： 目的分析下调IQ结构域三磷酸鸟苷酶激活蛋白(IQGAP)1对胰腺癌细胞增殖、凋亡的影响及作用机制。方法人胰腺癌HPAF-Ⅱ细胞分为对照组、过表达组、沉默组。对照组不做任何处理,过表达组为人胰腺癌细胞做IQGAP1过表达质粒转染,沉默组为人胰腺癌细胞做IQGAP1沉默质粒转染。鉴定IQGAP1转染效率,分析沉默IQGAP1基因对胰腺癌细胞的增殖抑制作用、凋亡及Wnt/β-连环蛋白(catenin)通路蛋白的影响,并分析其作用机制。结果过表达组IQGAP1基因表达量显著高于对照组,沉默组IQGAP1基因表达量显著低于对照组(P<0.05),说明IQGAP1基因转染成功。过表达组细胞增殖率显著高于对照组,沉默组细胞增殖率显著低于对照组、过表达组(P<0.05)。过表达组细胞凋亡率显著低于对照组,沉默组细胞凋亡率显著高于对照组、过表达组(P<0.05)。过表达组Wnt、β-catenin蛋白表达量显著低于对照组,细胞外信号调节激酶(ERK)、B细胞淋巴瘤-2相关X蛋白(Bax)、细胞外信号调节激酶的激酶(MEK)蛋白表达量显著高于对照组,沉默组Wnt、β-catenin蛋白表达量显著高于对照组,ERK、Bax、MEK蛋白表达量显著低于对照组(P<0.05);相比于过表达组,沉默组Wnt、β-catenin蛋白表达量显著较高,ERK、Bax、MEK蛋白表达量显著较低(P<0.05)。结论下调IQGAP1基因可以抑制胰腺癌细胞增殖,促进其凋亡,其机制可能与激活Wnt/β-catenin通路相关。

关键词： circAnk3   Glutamate receptors   IQ motif containing GTPase activating protein 1   miR-7080-3p   Psychiatric disorder   Synaptic plasticity  

摘要： BACKGROUND: Circular RNAs are highly enriched in the synapses of the mammalian brain and play important roles in neurological function by acting as molecular sponges of microRNAs. circAnk3 is derived from the 11th intron of the ankyrin-3 gene, Ank3 , a strong genetic risk factor for neuropsychiatric disorders;however, the function of circAnk3 remains elusive. In this study, we investigated the function of circAnk3 and its downstream regulatory network for target genes in the hippocampus of mice. METHODS: The DNA sequence from which circAnk3 is generated was modified using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/Cas9) technology, and neurobehavioral tests (anxiety and depression -like behaviors, social behaviors) were performed in circAnk3 + /- mice. A series of molecular and biochemical assays were used to investigate the function of circAnk3 as a microRNA sponge and its downstream regulatory network for target genes. RESULTS: circAnk3 + /- mice exhibited both anxiety -like behaviors and social deficits. circAnk3 was predominantly located in the cytoplasm of neuronal cells and functioned as a miR-7080-3p sponge to regulate the expression of Iqgap1 . Inhibition of miR-7080-3p or restoration of Iqgap1 in the hippocampus ameliorated the behavioral deficits of circAnk3 + /- mice. Furthermore, circAnk3 deficiency decreased the expression of the NMDA receptor subunit GluN2a and impaired the structural plasticity of dendritic synapses in the hippocampus. CONCLUSIONS: Our results reveal an important role of the circAnk3/miR-7080-3p/IQGAP1 axis in maintaining the structural plasticity of hippocampal synapses. circAnk3 might offer new insights into the involvement of circular RNAs in neuropsychiatric disorders.

关键词： cancer metastasis   Rho GTPases   endothelial adhesion   β-integrin   signaling   shear stress   IQGAP1   NWASP   cell-cell adhesion  

摘要： Attachment of circulating tumor cells to the endothelial cells (ECs) lining blood vessels is a critical step in cancer metastatic colonization, which leads to metastatic outgrowth. Breast and prostate cancers are common malignancies in women and men, respectively. Here, we observe that 01-integrin is required for human prostate and breast cancer cell adhesion to ECs under shear -stress conditions in vitro and to lung blood vessel ECs in vivo. We identify IQGAP1 and neural Wiskott-Aldrich syndrome protein (NWASP) as regulators of 01-integrin transcription and protein expression in prostate and breast cancer cells. IQGAP1 and NWASP depletion in cancer cells decreases adhesion to ECs in vitro and retention in the lung vasculature and metastatic lung nodule formation in vivo. Mechanistically, NWASP and IQGAP1 act downstream of Cdc42 to increase 01-integrin expression both via extracellular signal -regulated kinase (ERK)/focal adhesion kinase signaling at the protein level and by myocardin-related transcription factor/ serum response factor (SRF) transcriptionally. Our results identify IQGAP1 and NWASP as potential therapeutic targets to reduce early metastatic dissemination.

关键词： ANGPTL4   Cancer-associated fibroblasts   Chemosensitivity   Mitochondria   Prostate cancer  

摘要： Introduction: Cancer-associated fibroblasts (CAFs) are a critical component of the tumor microenvironment, being implicated in enhancing tumor growth and fostering drug resistance. Nonetheless, the mechanisms underlying their function in prostate cancer (PCa) remain incompletely understood, which is essential for devising effective therapeutic strategies. Objectives: The main objective of this study was to explore the mechanisms by which CAFs mediate PCa growth and chemoresistance. Methods: We validated through data analysis and experimentation that CAFs significantly impact PCa cell proliferation and chemoresistance. Subsequently, we conducted a comprehensive proteomic analysis of the conditioned media from CAFs and PCa cells and identified angiopoietin-like protein 4 (ANGPTL4) as a key factor. We employed ELISA and multiplex immunofluorescence assays, all of which indicated that ANGPTL4 was primarily secreted by ***, we conducted metabolomics analysis, GST pull-down assays, Co-IP, and other experiments to explore the specific molecular mechanisms of ANGPTL4 and its precise effects on PCa cells. Through drug screening, we identified Quercetin 3-O-(6′-galactopyranosyl)-β-D-galactopyranoside (QGGP) as an effective inhibitor of CAFs function. Finally, we thoroughly assessed the therapeutic potential of QGGP both as a monotherapy and in combination with docetaxel in PCa cells. Results: We discovered that the extracrine factor ANGPTL4 is primarily expressed in CAFs in PCa. When ANGPTL4 binds to IQ motif-containing GTPase-activating protein 1 (IQGAP1) on the PCa cell membrane, it activates the Raf-MEK-ERK-PGC1α axis, promoting mitochondrial biogenesis and OXPHOS metabolism, and thereby facilitating PCa growth and chemoresistance. Furthermore, virtual and functional screening strategies identified QGGP as a specific inhibitor of IQGAP1 that promotes its degradation. Combined with docetaxel treatment, QGGP can reverse the effects of CAFs and improve the responsivenes

关键词： 动脉粥样硬化   内皮细胞   细胞凋亡  

摘要： 背景：　　动脉粥样硬化（Atherosclerosis,AS）是一种慢性炎症性疾病，其导致的心脑血管疾病是人类健康所面对的最大威胁之一。正常血管内皮细胞可以调节血管张力、预防血栓形成和调节炎症，在维持血管稳态和抑制动脉粥样硬化方面具有重要作用，而脂质沉积诱导的内皮细胞损伤可以促进AS形成。目前，越来越多的证据表明，内皮细胞凋亡是动脉粥样硬化的始动因素。　　IQ结构域GTP酶激活蛋白1（IQmotifcontainingGTPaseactivatingproteins1,IQGAP1）是多种信号通路的重要调节因子，可以调控细胞生长、发育以及死亡，参与动脉粥样硬化的发生发展。另有研究表明IQGAP1可能调节Hippo信号通路的关键蛋白YAP（Yes-associatedprotein）从而影响细胞凋亡。所以本课题研究了IQGAP1在内皮细胞凋亡和动脉粥样硬化中的作用，并初步探索其作用机制。　　目的：　　研究IQGAP1对动脉粥样硬化中血管内皮细胞凋亡和YAP的调控作用。　　研究方法：　　1.通过高脂饮食建立小鼠动脉粥样硬化模型，应用WesternBlot方法鉴定IQGAP1以及凋亡相关蛋白在动脉粥样硬化小鼠模型中表达水平。　　2.体外构建棕榈酸（Palmiticacid,PA）诱导人脐静脉内皮细胞（HumanUmbilicalVeinEndothelialCells,HUVEC）凋亡模型，在凋亡模型中使用Jetprime转染小干扰RNA（SmallinterferingRNA,siRNA）敲低或者pcDNA3.1-3xFlag-IQGAP1过表达细胞内IQGAP1水平，采用流式细胞术（FlowCytometry,FCM）观察细胞凋亡并采用WB检测细胞凋亡蛋白改变水平。探索脂质沉积诱导的内皮细胞凋亡与IQGAP1的关系。　　3.在HUVEC中敲低或者过表达IQGAP1，使用WB及免疫荧光观察YAP蛋白水平的变化。　　4.在HUVEC中使用siRNA同时敲低IQGAP1和YAP，采用流式细胞术观察细胞凋亡并采用WB检测细胞凋亡蛋白改变水平。　　5.通过AAV-siRNA在动脉粥样硬化模型小鼠中实现敲低IQGAP1，进一步探索IQGAP1在动脉粥样硬化模型鼠血管内皮凋亡中发挥的作用。　　结果：　　1.通过高脂饮食建立小鼠动脉粥样硬化模型，WB检测到高脂饮食小鼠主动脉中IQGAP1，Caspase-3，BAX（Bcl-2-associatedXprotein）高于对照组，BCL-2低于对照组。TUNEL染色结果表明高脂饮食小鼠主动脉凋亡细胞高于对照组。　　2.在PA诱导的HUVEC凋亡模型中，敲低IQGAP1之后，Caspase-3和BAX水平降低，BCL-2（B-celllymphoma-2）升高;反之上调。流式细胞术表明，敲低IQGAP1后，细胞凋亡水平有所降低;反之升高。EdU染色结果表明，敲低IQGAP1后，细胞增殖水平有所升高;反之降低。　　3.在HUVEC细胞中敲低IQGAP1后，免疫荧光及WB结果表明YAP蛋白表达量有所升高;反之，过表达IQGAP1后，YAP蛋白表达量降低。　　4.在HUVEC细胞中敲低IQGAP1后，流式细胞术表明细胞凋亡水平降低，但将IQGAP1及YAP同时敲低后，细胞凋亡水平相较于单独敲低IQGAP1升高。敲低YAP逆转了敲低IQGAP1带来的抗凋亡作用。　　5.在高脂饮食小鼠体内注射AAV-si-IQGAP1后，主动脉表达IGQAP1降低，动脉粥样硬化斑块大小降低，细胞凋亡水平降低，且YAP升高。　　结论：　　1.动脉粥样硬化斑块中凋亡水平高于正常组，IQGAP1在动脉粥样硬化斑块中表达明显增高。　　2.使用siRNA敲低IQGAP1可以抑制PA诱导的细胞凋亡，并促进细胞增殖。反之过表达IQGAP1会诱导细胞发生凋亡，抑制细胞增殖。　　***的表达水平与IQGAP1呈负相关，敲低IQGAP1会增加YAP的表达量，而过表达IQGAP1会降低YAP的表达量。　　4.使用siRNA敲低YAP后，可以促进细胞发生凋亡，抑制细胞增殖。　　5.在小鼠高脂饮食模型中使用AAV-si-RNA，验证了敲低IQGAP1可以降低小鼠主动脉粥样硬化斑块和细胞凋亡水平，敲低IQGAP1后YAP表达水平升高。