Background:The tumor microenvironment(TME)plays an important role in regulating gastric cancer(GC)progression.The infiltration of M0 macrophages into the TME negatively affects the prognosis of patients with various t...Background:The tumor microenvironment(TME)plays an important role in regulating gastric cancer(GC)progression.The infiltration of M0 macrophages into the TME negatively affects the prognosis of patients with various tumors.Methods:The data were obtained from the TCGA and GEO databases and our hospital(107 patients).The expression of IQGAP3 was knocked down in AGS and NCI-N87 GC cells.Cell proliferation,migration,and invasion assays were performed.RNA sequencing was performed on GC cells with different IQGAP3 expression.AGS and THP-1 cells were mixed to create a co-cultured subcutaneous tumor model in nude mice.Tumor growth in the mice was observed by using luciferin fluorescence and the tumor tissues were subjected to immunohistochemistry.Results:The expression of IQGAP3 was increased in GC tissues(P<0.001)and was associated with infiltration of M0 macrophages and a poor prognosis for GC patients(P<0.01).Knockdown of IQGAP3 resulted in decreased expression of CXCL13(P<0.001)and less phosphorylation of pSTAT3 and pERK1/2(P<0.001).The expression of CXCL13 was decreased after the pSTAT3 and pERK1/2 phosphorylation inhibitors were added.In the co-culture experiment,the M0/THP-1 ratio decreased significantly in the low-IQGAP3-expression group(P<0.001).However,adding recombinant human CXCL13 proteins to the low IQGAP3 expression group increased the M0/THP-1 ratio.In vivo,tumor growth and M0 macrophage infiltration were both suppressed in the group with low IQGAP3 expression.Conclusion:IQGAP3 is a potential pro-carcinogenic factor in GC.IQGAP3 promotes the expression and secretion of CXCL13 via the ERK1/2 and STAT3 pathways,thereby causing M0 macrophages to infiltrate the TME.展开更多
Background:IQGAP3 plays a crucial role in regulating cell proliferation,division,and cytoskeletal organization.Abnormal expression of IQGAP3 has been linked to various tumors,but its function in glioma is not well und...Background:IQGAP3 plays a crucial role in regulating cell proliferation,division,and cytoskeletal organization.Abnormal expression of IQGAP3 has been linked to various tumors,but its function in glioma is not well understood.Methods:Various methods,including genetic differential analysis,single-cell analysis,ROC curve analysis,Cox regression,Kaplan-Meier analysis,and enrichment analysis,were employed to analyze the expression patterns,diagnostic potential,prognostic implications,and biological processes involving IQGAP3 in normal and tumor tissues.The impact of IQGAP3 on immune infiltration and the immune microenvironment in gliomas was evaluated using immunofluorescence.Additionally,the cBioPortal database was used to analyze copy number variations and mutation sites of IQGAP3.Experimental validation was also performed to assess the effects of IQGAP3 on glioma cells and explore underlying mechanisms.Results:High IQGAP3 expression in gliomas is associated with an unfavorable prognosis,particularly in wild-type IDH and 1p/19q non-codeleted gliomas.Enrichment analysis revealed that IQGAP3 is involved in regulating the cell cycle,PI3K/AKT signaling,p53 signaling,and PLK1-related pathways.Furthermore,IQGAP3 expression may be closely related to the immunosuppressive microenvironment of glioblastoma.BRD-K88742110 and LY-303511 are potential drugs for targeting IQGAP3 in anti-glioma therapy.In vitro experiments showed that downregulation of IQGAP3 inhibits the proliferation and migration of glioma cells,with the PLK1/PI3K/AKT pathway potentially playing a crucial role in IQGAP3-mediated glioma progression.Conclusion:IQGAP3 shows promise as a valuable biomarker for diagnosis,prognosis,and immunotherapeutic strategies in gliomas.展开更多
目的探讨miR-603抑制宫颈癌细胞增殖和迁移的机制。方法采用基因编辑系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9,CRISPR/Cas9)分别敲除宫颈癌细胞SiHa中人乳头瘤病毒16(human papil...目的探讨miR-603抑制宫颈癌细胞增殖和迁移的机制。方法采用基因编辑系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9,CRISPR/Cas9)分别敲除宫颈癌细胞SiHa中人乳头瘤病毒16(human papilloma virus 16,HPV16)E6、E7后采用real-time PCR检测miR-603的表达。将宫颈癌细胞SiHa分为4组:mimic NC组、mimic 603组、inhibitor NC组和inhibitor 603组,将mimic NC、mimic 603、inhibitor NC及inhibitor 603分别转染SiHa细胞,通过real-time PCR检测miR-603的表达水平,CCK-8法检测SiHa细胞增殖能力,Transwell小室法检测SiHa细胞迁移能力。双荧光素酶报告试验检测miR-603是否与IQ结构域GTP酶激活蛋白3(IQ motif-containing GTPase activating protein 3,IQGAP3)直接结合,Western blotting检测IQGAP3蛋白表达。UALCAN(The University of Alabama at Birmingham cancer data analysis portal)数据库分析IQGAP3在肿瘤中的表达情况;通过肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库分析IQGAP3在宫颈癌细胞系中的表达;利用分析癌症基因表达和病毒感染的交互式在线数据库(an interactive online database for analysis of gene expression and viral infection in cancer,OncoDB)分析IQGAP3与宫颈癌患者HPV感染的相关性。结果敲除HPV16 E6、E7后miR-603的表达升高(P<0.05)。过表达miR-603后,SiHa细胞的增殖和迁移能力下降(P<0.05);敲低miR-603后,SiHa细胞的增殖和迁移能力增强(P<0.05)。双荧光素酶报告试验提示IQGAP3是miR-603的直接靶点,过表达miR-603后,IQGAP3的表达降低,敲低miR-603可促进IQGAP3的表达。UALCAN数据库显示IQGAP3在多种肿瘤组织中表达高于正常对照,OncoDB数据库分析发现HPV16、18阳性患者IQGAP3表达均高于阴性患者。使用CCLE数据库分析IQGAP3在宫颈癌细胞系的表达,发现HPV阴性C33A细胞中IQGAP3表达明显低于HPV16阳性CASKI、SiHa细胞,以及HPV18阳性HELA、C4I、C4II及MS751细胞。结论敲除HPV16 E6、E7后miR-603表达上调,miR-603可抑制宫颈癌SiHa细胞的增殖和迁移,这可能与其直接靶向宫颈癌中高表达的IQGAP3有关。展开更多
基金National Natural Science Foundation of China(no.82372860,no.81641098)Peking University First Hospital Youth Clinical Research Project(no.2021CR03)+3 种基金Peking University First Hospital Multidisciplinary Research Project(no.2022CR13)the Youth Cultivated Research Fund of Peking University Health Center(grant no.BMU2020PYB026)National Project for Clinical Key Specialty DevelopmentTianjin Key Medical Discipline Construction Project(grant no.TJYXZDXK-3-003A).
文摘Background:The tumor microenvironment(TME)plays an important role in regulating gastric cancer(GC)progression.The infiltration of M0 macrophages into the TME negatively affects the prognosis of patients with various tumors.Methods:The data were obtained from the TCGA and GEO databases and our hospital(107 patients).The expression of IQGAP3 was knocked down in AGS and NCI-N87 GC cells.Cell proliferation,migration,and invasion assays were performed.RNA sequencing was performed on GC cells with different IQGAP3 expression.AGS and THP-1 cells were mixed to create a co-cultured subcutaneous tumor model in nude mice.Tumor growth in the mice was observed by using luciferin fluorescence and the tumor tissues were subjected to immunohistochemistry.Results:The expression of IQGAP3 was increased in GC tissues(P<0.001)and was associated with infiltration of M0 macrophages and a poor prognosis for GC patients(P<0.01).Knockdown of IQGAP3 resulted in decreased expression of CXCL13(P<0.001)and less phosphorylation of pSTAT3 and pERK1/2(P<0.001).The expression of CXCL13 was decreased after the pSTAT3 and pERK1/2 phosphorylation inhibitors were added.In the co-culture experiment,the M0/THP-1 ratio decreased significantly in the low-IQGAP3-expression group(P<0.001).However,adding recombinant human CXCL13 proteins to the low IQGAP3 expression group increased the M0/THP-1 ratio.In vivo,tumor growth and M0 macrophage infiltration were both suppressed in the group with low IQGAP3 expression.Conclusion:IQGAP3 is a potential pro-carcinogenic factor in GC.IQGAP3 promotes the expression and secretion of CXCL13 via the ERK1/2 and STAT3 pathways,thereby causing M0 macrophages to infiltrate the TME.
基金supported by the Doctoral Foundation of HuBei University of Science and Technology(Grant Numbers BK202007 and BK202028 to L.W.and Z.Z.)Special Research Fund Project of School of Stomatology and Optometry,Xianning Medical College,Hubei University of Science and Technology(Grant Number 2020XZ37 to L.W.)+3 种基金Hubei Provincial Department of Education“Hundred Schools and Hundred Counties”(Grant Number BXLBX0806 to Z.Z.)the Foundation of Hubei University of Science and Technology“Double Hundred Project”(Grant Number 2022HKSB01 to Z.Z.)the Foundation of Innovation Team of Hubei University of Science and Technology(Grant Number 2023T13 to S.Y.)Natural Science Foundation of Hubei Province(Grant Number 2023AFB1027 to Z.Z.).
文摘Background:IQGAP3 plays a crucial role in regulating cell proliferation,division,and cytoskeletal organization.Abnormal expression of IQGAP3 has been linked to various tumors,but its function in glioma is not well understood.Methods:Various methods,including genetic differential analysis,single-cell analysis,ROC curve analysis,Cox regression,Kaplan-Meier analysis,and enrichment analysis,were employed to analyze the expression patterns,diagnostic potential,prognostic implications,and biological processes involving IQGAP3 in normal and tumor tissues.The impact of IQGAP3 on immune infiltration and the immune microenvironment in gliomas was evaluated using immunofluorescence.Additionally,the cBioPortal database was used to analyze copy number variations and mutation sites of IQGAP3.Experimental validation was also performed to assess the effects of IQGAP3 on glioma cells and explore underlying mechanisms.Results:High IQGAP3 expression in gliomas is associated with an unfavorable prognosis,particularly in wild-type IDH and 1p/19q non-codeleted gliomas.Enrichment analysis revealed that IQGAP3 is involved in regulating the cell cycle,PI3K/AKT signaling,p53 signaling,and PLK1-related pathways.Furthermore,IQGAP3 expression may be closely related to the immunosuppressive microenvironment of glioblastoma.BRD-K88742110 and LY-303511 are potential drugs for targeting IQGAP3 in anti-glioma therapy.In vitro experiments showed that downregulation of IQGAP3 inhibits the proliferation and migration of glioma cells,with the PLK1/PI3K/AKT pathway potentially playing a crucial role in IQGAP3-mediated glioma progression.Conclusion:IQGAP3 shows promise as a valuable biomarker for diagnosis,prognosis,and immunotherapeutic strategies in gliomas.
文摘目的探讨miR-603抑制宫颈癌细胞增殖和迁移的机制。方法采用基因编辑系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9,CRISPR/Cas9)分别敲除宫颈癌细胞SiHa中人乳头瘤病毒16(human papilloma virus 16,HPV16)E6、E7后采用real-time PCR检测miR-603的表达。将宫颈癌细胞SiHa分为4组:mimic NC组、mimic 603组、inhibitor NC组和inhibitor 603组,将mimic NC、mimic 603、inhibitor NC及inhibitor 603分别转染SiHa细胞,通过real-time PCR检测miR-603的表达水平,CCK-8法检测SiHa细胞增殖能力,Transwell小室法检测SiHa细胞迁移能力。双荧光素酶报告试验检测miR-603是否与IQ结构域GTP酶激活蛋白3(IQ motif-containing GTPase activating protein 3,IQGAP3)直接结合,Western blotting检测IQGAP3蛋白表达。UALCAN(The University of Alabama at Birmingham cancer data analysis portal)数据库分析IQGAP3在肿瘤中的表达情况;通过肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库分析IQGAP3在宫颈癌细胞系中的表达;利用分析癌症基因表达和病毒感染的交互式在线数据库(an interactive online database for analysis of gene expression and viral infection in cancer,OncoDB)分析IQGAP3与宫颈癌患者HPV感染的相关性。结果敲除HPV16 E6、E7后miR-603的表达升高(P<0.05)。过表达miR-603后,SiHa细胞的增殖和迁移能力下降(P<0.05);敲低miR-603后,SiHa细胞的增殖和迁移能力增强(P<0.05)。双荧光素酶报告试验提示IQGAP3是miR-603的直接靶点,过表达miR-603后,IQGAP3的表达降低,敲低miR-603可促进IQGAP3的表达。UALCAN数据库显示IQGAP3在多种肿瘤组织中表达高于正常对照,OncoDB数据库分析发现HPV16、18阳性患者IQGAP3表达均高于阴性患者。使用CCLE数据库分析IQGAP3在宫颈癌细胞系的表达,发现HPV阴性C33A细胞中IQGAP3表达明显低于HPV16阳性CASKI、SiHa细胞,以及HPV18阳性HELA、C4I、C4II及MS751细胞。结论敲除HPV16 E6、E7后miR-603表达上调,miR-603可抑制宫颈癌SiHa细胞的增殖和迁移,这可能与其直接靶向宫颈癌中高表达的IQGAP3有关。
