目的利用生物信息分析FZDs基因家族(FZD1~FZD10)在原发性肝癌中的作用。方法肝癌样本mRNA表达数据取自The Cancer Genome Atlas(TCGA),采用仙桃学术、GEPIA数据库、Kaplan-Meier Plotter数据库、cBioPortal数据库全面分析FZDs基因家族...目的利用生物信息分析FZDs基因家族(FZD1~FZD10)在原发性肝癌中的作用。方法肝癌样本mRNA表达数据取自The Cancer Genome Atlas(TCGA),采用仙桃学术、GEPIA数据库、Kaplan-Meier Plotter数据库、cBioPortal数据库全面分析FZDs基因家族在原发性肝癌中的表达及表达与肝癌分期和预后的关系,并进一步分析其基因突变水平和相关基因的通路富集。结果在肝癌组织中FZD1、FZD8的mRNA表达水平降低(P<0.05),FZD2、FZD3、FZD4、FZD5、FZD6、FZD10表达水平升高(P<0.05),且FZDs基因家族成员之间表达存在相关性。FZD1、FZD2、FZD6、FZD7、FZD8、FZD10表达与肝癌分期密切相关(P<0.05)。FZD1、FZD5、FZD6、FZD7高表达,FZD4、FZD9、FZD10低表达与肝癌患者预后不良相关(P<0.05)。此外,原发性肝癌中FZDs基因家族的总突变频率为29.78%,且所有基因均有突变发生。GO和KEGG富集分析发现FZDs基因家族在原发性肝癌作用的机制可能与Wnt信号通路、Hippo信号通路、干细胞多能干性调控相关信号通路和mTOR信号通路有关。结论FZDs基因家族成员可作为肝癌潜在治疗靶点和预后预测标记物,在原发性肝癌发生发展中起着关键作用。展开更多
Objective:In the realm of Class I pathogens,Burkholderia pseudomallei(BP)stands out for its propensity to induce severe pathogenicity.Investigating the intricate interactions between BP and host cells is imperative fo...Objective:In the realm of Class I pathogens,Burkholderia pseudomallei(BP)stands out for its propensity to induce severe pathogenicity.Investigating the intricate interactions between BP and host cells is imperative for comprehending the dynamics of BP infection and discerning biomarkers indicative of the host cell response process.Methods:mRNA extraction from BP-infected mouse macrophages constituted the initial step of our study.Employing gene expression arrays,the extracted RNA underwent conversion into digital signals.The percentile shift method facilitated data processing,with the identification of genes manifesting significant differences accomplished through the application of the t-test.Subsequently,a comprehensive analysis involving Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway was conducted on the differentially expressed genes(DEGs).Leveraging the ESTIMATE algorithm,gene signatures were utilized to compute risk scores for gene expression data.Support vector machine analysis and gene enrichment scores were instrumental in establishing correlations between biomarkers and macrophages,followed by an evaluation of the predictive power of the identified biomarkers.Results:The functional and pathway associations of the DEGs predominantly centered around G protein-coupled receptors.A noteworthy positive correlation emerged between the blue module,consisting of 416 genes,and the StromaScore.FZD4,identified through support vector machine analysis among intersecting genes,indicated a robust interaction with macrophages,suggesting its potential as a robust biomarker.FZD4 exhibited commendable predictive efficacy,with BP infection inducing its expression in both macrophages and mouse lung tissue.Western blotting in macrophages confirmed a significant upregulation of FZD4 expression from 0.5 to 24 h post-infection.In mouse lung tissue,FZD4 manifested higher expression in the cytoplasm of pulmonary epithelial cells in BP-infected lungs than in the control group.Conclusion:Thesefindings underscore the upregulation of FZD4 expression by BP in both macrophages and lung tissue,pointing to its prospective role as a biomarker in the pathogenesis of BP infection.展开更多
BACKGROUND Wnt/FZD-mediated signaling pathways are activated in more than 90%of hepatocellular carcinoma(HCC)cell lines.As a well-known secretory glycoprotein,Wnt3 can interact with FZD receptors on the cell surface,t...BACKGROUND Wnt/FZD-mediated signaling pathways are activated in more than 90%of hepatocellular carcinoma(HCC)cell lines.As a well-known secretory glycoprotein,Wnt3 can interact with FZD receptors on the cell surface,thereby activating the Wnt/β-catenin signaling pathway.However,the N-glycosylation modification site of Wnt3 and the effect of this modification on the biological function of the protein are still unclear.AIM To investigate the effect of Wnt3 N-glycosylation on the biological function of HCC cells.METHODS Site-directed mutagenesis was used to verify the Wnt3 N-glycosylation sites,actinomycin D treatment was used to detect the stability of Wnt3 after site-directed mutation,the binding of the N-glycosylation site-directed mutant Wnt3 to FZD7 was observed by laser confocal microscopy,and the effects of the N-glycosylation site-directed mutation of Wnt3 on the Wnt/β-catenin signaling pathway and the progression of HCC cells were detected by western blot and cell function experiments.RESULTS Wnt3 has two N-glycosylation-modified sites(Asn90 and Asn301);when a single site at amino acid 301 is mutated,the stability of Wnt3 is weakened;the binding ability of Wnt3 to FZD7 decreases when both sites are mutated simultaneously;and the level of proteins related to the Wnt/β-catenin signaling pathway is downregulated.Cell proliferation,migration and invasion are also weakened in the case of single 301 site and double-site mutations.CONCLUSION These results indicate that by inhibiting the N-glycosylation of Wnt3,the proliferation,migration,invasion and colony formation abilities of liver cancer cells can be weakened,which might provide new therapeutic strategies for clinical liver cancer in the future.展开更多
文摘目的利用生物信息分析FZDs基因家族(FZD1~FZD10)在原发性肝癌中的作用。方法肝癌样本mRNA表达数据取自The Cancer Genome Atlas(TCGA),采用仙桃学术、GEPIA数据库、Kaplan-Meier Plotter数据库、cBioPortal数据库全面分析FZDs基因家族在原发性肝癌中的表达及表达与肝癌分期和预后的关系,并进一步分析其基因突变水平和相关基因的通路富集。结果在肝癌组织中FZD1、FZD8的mRNA表达水平降低(P<0.05),FZD2、FZD3、FZD4、FZD5、FZD6、FZD10表达水平升高(P<0.05),且FZDs基因家族成员之间表达存在相关性。FZD1、FZD2、FZD6、FZD7、FZD8、FZD10表达与肝癌分期密切相关(P<0.05)。FZD1、FZD5、FZD6、FZD7高表达,FZD4、FZD9、FZD10低表达与肝癌患者预后不良相关(P<0.05)。此外,原发性肝癌中FZDs基因家族的总突变频率为29.78%,且所有基因均有突变发生。GO和KEGG富集分析发现FZDs基因家族在原发性肝癌作用的机制可能与Wnt信号通路、Hippo信号通路、干细胞多能干性调控相关信号通路和mTOR信号通路有关。结论FZDs基因家族成员可作为肝癌潜在治疗靶点和预后预测标记物,在原发性肝癌发生发展中起着关键作用。
基金The study was supported by Yuying Program Incubation Project of General Hospital of Center Theater(ZZYFH202104)Wuhan Young and Middle-Aged Medical Backbone Talent Project 2020(2020-55)Logistics Research Program Project 2019(CLB19J029).
文摘Objective:In the realm of Class I pathogens,Burkholderia pseudomallei(BP)stands out for its propensity to induce severe pathogenicity.Investigating the intricate interactions between BP and host cells is imperative for comprehending the dynamics of BP infection and discerning biomarkers indicative of the host cell response process.Methods:mRNA extraction from BP-infected mouse macrophages constituted the initial step of our study.Employing gene expression arrays,the extracted RNA underwent conversion into digital signals.The percentile shift method facilitated data processing,with the identification of genes manifesting significant differences accomplished through the application of the t-test.Subsequently,a comprehensive analysis involving Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway was conducted on the differentially expressed genes(DEGs).Leveraging the ESTIMATE algorithm,gene signatures were utilized to compute risk scores for gene expression data.Support vector machine analysis and gene enrichment scores were instrumental in establishing correlations between biomarkers and macrophages,followed by an evaluation of the predictive power of the identified biomarkers.Results:The functional and pathway associations of the DEGs predominantly centered around G protein-coupled receptors.A noteworthy positive correlation emerged between the blue module,consisting of 416 genes,and the StromaScore.FZD4,identified through support vector machine analysis among intersecting genes,indicated a robust interaction with macrophages,suggesting its potential as a robust biomarker.FZD4 exhibited commendable predictive efficacy,with BP infection inducing its expression in both macrophages and mouse lung tissue.Western blotting in macrophages confirmed a significant upregulation of FZD4 expression from 0.5 to 24 h post-infection.In mouse lung tissue,FZD4 manifested higher expression in the cytoplasm of pulmonary epithelial cells in BP-infected lungs than in the control group.Conclusion:Thesefindings underscore the upregulation of FZD4 expression by BP in both macrophages and lung tissue,pointing to its prospective role as a biomarker in the pathogenesis of BP infection.
基金Supported by National Natural Science Foundation of China,No.81560390the Guizhou Medical University Cultivation Project of the National Natural Science Foundation of China,No.22NSFCP02Basic Research Project of Science and Technology Department of Guizhou Province,No.ZK[2024]General 136.
文摘BACKGROUND Wnt/FZD-mediated signaling pathways are activated in more than 90%of hepatocellular carcinoma(HCC)cell lines.As a well-known secretory glycoprotein,Wnt3 can interact with FZD receptors on the cell surface,thereby activating the Wnt/β-catenin signaling pathway.However,the N-glycosylation modification site of Wnt3 and the effect of this modification on the biological function of the protein are still unclear.AIM To investigate the effect of Wnt3 N-glycosylation on the biological function of HCC cells.METHODS Site-directed mutagenesis was used to verify the Wnt3 N-glycosylation sites,actinomycin D treatment was used to detect the stability of Wnt3 after site-directed mutation,the binding of the N-glycosylation site-directed mutant Wnt3 to FZD7 was observed by laser confocal microscopy,and the effects of the N-glycosylation site-directed mutation of Wnt3 on the Wnt/β-catenin signaling pathway and the progression of HCC cells were detected by western blot and cell function experiments.RESULTS Wnt3 has two N-glycosylation-modified sites(Asn90 and Asn301);when a single site at amino acid 301 is mutated,the stability of Wnt3 is weakened;the binding ability of Wnt3 to FZD7 decreases when both sites are mutated simultaneously;and the level of proteins related to the Wnt/β-catenin signaling pathway is downregulated.Cell proliferation,migration and invasion are also weakened in the case of single 301 site and double-site mutations.CONCLUSION These results indicate that by inhibiting the N-glycosylation of Wnt3,the proliferation,migration,invasion and colony formation abilities of liver cancer cells can be weakened,which might provide new therapeutic strategies for clinical liver cancer in the future.
