目的探讨接头相关复合体蛋白3亚基2(adaptor related protein complex 3 subunit mu 2,AP3M2)在胃癌中的表达情况及其临床意义。方法通过TCGA数据库、实时荧光定量PCR及免疫印迹实验检测AP3M2基因的mRNA及蛋白在胃癌组织中的表达情况。...目的探讨接头相关复合体蛋白3亚基2(adaptor related protein complex 3 subunit mu 2,AP3M2)在胃癌中的表达情况及其临床意义。方法通过TCGA数据库、实时荧光定量PCR及免疫印迹实验检测AP3M2基因的mRNA及蛋白在胃癌组织中的表达情况。通过检测KM Plotter网站探索AP3M2的表达与胃癌患者预后的关系。通过平板克隆形成实验检测AP3M2对胃癌细胞的作用。结果TCGA数据库数据及本研究收集的胃癌患者组织显示AP3M2相比于癌旁组织,其在胃癌组织中的mRNA表达水平升高,同时AP3M2的蛋白在胃癌组织中的表达也有上调的现象。同时本研究通过KM Plotter网站分析得到AP3M2的mRNA表达水平与胃癌患者的预后有关,高表达AP3M2的胃癌患者其预后较低表达AP3M2的胃癌患者差。通过在AGS细胞(人胃腺癌细胞)中过表达AP3M2并检测其平板克隆形成能力的变化,发现过表达AP3M2可以增强胃癌细胞的克隆形成能力。结论AP3M2在胃癌中高表达,高表达AP3M2与胃癌患者的不良预后有关,且能增强胃癌细胞的克隆形成能力。展开更多
Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its g...Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its gene expression level and prognosis.Methods:The exosomes of hepatocellular carcinoma cells HepG2 and 7721 were isolated by ultracentrifugation,and the extracted exosomes were verified by transmission electron microscope, western blotting,and nanosight.The relative quantitative proteomics analysis was used to analyze the protein spectrum of HepG2 and 7721 cells in exosomes.The bioinformatics was used to collect the signal pathway,and the target protein AP2M1 was verified by qRTPCR.Gene Expression Profiling Interactive Analysis(GEPIA),OncoLnc,and The Cancer Genome Atlas(TCGA)database were used to analyze the expression level of AP2M1 and the survival rate of patients with liver cancer.Results:The extracellular exosomes of HepG2 and 7721 cells were isolated by ultracentrifugation.High-purity exosomes were obtained and verified by transmission electron microscopy,western blotting,and nanosight.There were 836 proteins co-expressed by the exosomes of HepG2 and 7721.By the analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,a total of 34 pathways that were related to liver cancer were collected,and the target membrane protein AP2M1 was screened.AP2M1 highly expressed in HepG2 cells,HepG2 and Huh7 cells' exosomes were detected by qRT-PCR(P<0.05).The data of GEPIA showed that comparing to the adjacent tissues,gene AP2M1 was highly expressed in hepatocarcinoma tissues.Kaplan-Meier survival curve analysis was performed by OncoLnc.It was found that the survival rate of patients with liver cancer and high expression of AP2M1 was significantly lower than patients with liver cancer and low expression of AP2M1(P<0.01).Conclusion:The high expression of membrane protein AP2M1 in the exosomes of hepatoma cells and the high expression of its gene in liver cancer tissues predicted the low survival rate.展开更多
文摘目的探讨接头相关复合体蛋白3亚基2(adaptor related protein complex 3 subunit mu 2,AP3M2)在胃癌中的表达情况及其临床意义。方法通过TCGA数据库、实时荧光定量PCR及免疫印迹实验检测AP3M2基因的mRNA及蛋白在胃癌组织中的表达情况。通过检测KM Plotter网站探索AP3M2的表达与胃癌患者预后的关系。通过平板克隆形成实验检测AP3M2对胃癌细胞的作用。结果TCGA数据库数据及本研究收集的胃癌患者组织显示AP3M2相比于癌旁组织,其在胃癌组织中的mRNA表达水平升高,同时AP3M2的蛋白在胃癌组织中的表达也有上调的现象。同时本研究通过KM Plotter网站分析得到AP3M2的mRNA表达水平与胃癌患者的预后有关,高表达AP3M2的胃癌患者其预后较低表达AP3M2的胃癌患者差。通过在AGS细胞(人胃腺癌细胞)中过表达AP3M2并检测其平板克隆形成能力的变化,发现过表达AP3M2可以增强胃癌细胞的克隆形成能力。结论AP3M2在胃癌中高表达,高表达AP3M2与胃癌患者的不良预后有关,且能增强胃癌细胞的克隆形成能力。
基金supported by grants from the National Natural Science Foundation of China(No.81760612)Guangxi Key Research and Development Project(No.AB16380351)
文摘Objective:Adopting mass spectrometry to analyze the protein spectrum of the exosomes in human hepatocellular carcinoma cells and screening the target membrane protein in order to analyze the relationship between its gene expression level and prognosis.Methods:The exosomes of hepatocellular carcinoma cells HepG2 and 7721 were isolated by ultracentrifugation,and the extracted exosomes were verified by transmission electron microscope, western blotting,and nanosight.The relative quantitative proteomics analysis was used to analyze the protein spectrum of HepG2 and 7721 cells in exosomes.The bioinformatics was used to collect the signal pathway,and the target protein AP2M1 was verified by qRTPCR.Gene Expression Profiling Interactive Analysis(GEPIA),OncoLnc,and The Cancer Genome Atlas(TCGA)database were used to analyze the expression level of AP2M1 and the survival rate of patients with liver cancer.Results:The extracellular exosomes of HepG2 and 7721 cells were isolated by ultracentrifugation.High-purity exosomes were obtained and verified by transmission electron microscopy,western blotting,and nanosight.There were 836 proteins co-expressed by the exosomes of HepG2 and 7721.By the analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,a total of 34 pathways that were related to liver cancer were collected,and the target membrane protein AP2M1 was screened.AP2M1 highly expressed in HepG2 cells,HepG2 and Huh7 cells' exosomes were detected by qRT-PCR(P<0.05).The data of GEPIA showed that comparing to the adjacent tissues,gene AP2M1 was highly expressed in hepatocarcinoma tissues.Kaplan-Meier survival curve analysis was performed by OncoLnc.It was found that the survival rate of patients with liver cancer and high expression of AP2M1 was significantly lower than patients with liver cancer and low expression of AP2M1(P<0.01).Conclusion:The high expression of membrane protein AP2M1 in the exosomes of hepatoma cells and the high expression of its gene in liver cancer tissues predicted the low survival rate.
