目的:应用生物信息学方法和实验验证确定转录因子TFAP2C在人类膀胱癌中的作用。方法:通过TCGA、Oncomine、GEPIA、The Human Protein Altas和Kaplan-Meier Plotter等数据库获得膀胱癌患者TFAP2C的转录信息和生存数据,分析TFAP2C在膀胱...目的:应用生物信息学方法和实验验证确定转录因子TFAP2C在人类膀胱癌中的作用。方法:通过TCGA、Oncomine、GEPIA、The Human Protein Altas和Kaplan-Meier Plotter等数据库获得膀胱癌患者TFAP2C的转录信息和生存数据,分析TFAP2C在膀胱癌组织中的表达水平及与预后的关系。在si-TFAP2C转染膀胱癌5637细胞后,利用CCK8和划痕实验检验TFAP2C在膀胱癌细胞中的作用。用STRING数据库构建蛋白互作网络,用R软件对网络中的基因进行GO和KEGG富集分析,通过R软件将TFAP2C在TCGA膀胱癌样本中的表达情况进行GSEA富集分析并作免疫细胞浸润相关性分析。结果:TFAP2C在膀胱癌组织中高表达,且其高表达预示着膀胱癌患者的总体生存率较差(P<0.05)。划痕和CCK8实验证明了TFAP2C可促进膀胱癌细胞的增殖和迁移(P<0.05)。GSEA结果显示,TFAP2C的高表达样本富集于蛋白质分泌、有丝分裂纺锤体、PI3K/AKT/mTOR信号通路和mTORC1信号通路(FDR<0.1,|NES|>1,P<0.05)。KEGG信号通路分析显示:TFAP2C与相关基因主要通路富集于ErbB信号通路、EGFR酪氨酸激酶抑制剂耐药性和膀胱癌等(P<0.05)。GO功能富集分析显示:TFAP2C与相关基因的生物学过程主要富集于转录共激活因子活性、转录辅助调节因子活性和表皮生长因子受体结合等(P<0.05)。结论:TFAP2C在膀胱癌中起促癌作用,表达水平上调和预后不良有关,TFAP2C可能是判断膀胱癌预后的生物标志物。展开更多
Chitosan and its degradation product,oligosaccharides,have been shown to facilitate peripheral nerve regeneration.However,the underlying mechanisms are not well understood.In this study,we analyzed the protein express...Chitosan and its degradation product,oligosaccharides,have been shown to facilitate peripheral nerve regeneration.However,the underlying mechanisms are not well understood.In this study,we analyzed the protein expression profiles in sciatic nerves after injury using proteomics.A group of proteins related to exosome packaging and transport is up-regulated by chitosan oligosaccharides(COS),implying that exosomes are involved in COS-induced peripheral nerve regeneration.In fact,exosomes derived from fibroblasts(f-EXOs)treated with COS significantly promoted axon extension and regeneration.Exosomal protein identification and functional studies,revealed that TFAP2C is a key factor in neurite outgrowth induced by COS-f-EXOs.Furthermore,we showed that TFAP2C targets the pri-miRNA-132 gene and represses miR-132-5p expression in dorsal root ganglion neurons.Camkk1 is a downstream substrate of miR-132-5p that positively affects axon extension.In rats,miR-132-5p antagomir stimulates CAMKK1 expression and improves axon regeneration and functional recovery in sciatic nerves after injury.Our data reveal the mechanism for COS in axon regeneration,that is COS induce fibroblasts to produce TFAP2C-enriched EXOs,which are then transferred into axons to promote axon regeneration via miR-132-5p/CAMKK1.Moreover,these results show a new facet of fibroblasts in axon regeneration in peripheral nerves.展开更多
文摘目的:应用生物信息学方法和实验验证确定转录因子TFAP2C在人类膀胱癌中的作用。方法:通过TCGA、Oncomine、GEPIA、The Human Protein Altas和Kaplan-Meier Plotter等数据库获得膀胱癌患者TFAP2C的转录信息和生存数据,分析TFAP2C在膀胱癌组织中的表达水平及与预后的关系。在si-TFAP2C转染膀胱癌5637细胞后,利用CCK8和划痕实验检验TFAP2C在膀胱癌细胞中的作用。用STRING数据库构建蛋白互作网络,用R软件对网络中的基因进行GO和KEGG富集分析,通过R软件将TFAP2C在TCGA膀胱癌样本中的表达情况进行GSEA富集分析并作免疫细胞浸润相关性分析。结果:TFAP2C在膀胱癌组织中高表达,且其高表达预示着膀胱癌患者的总体生存率较差(P<0.05)。划痕和CCK8实验证明了TFAP2C可促进膀胱癌细胞的增殖和迁移(P<0.05)。GSEA结果显示,TFAP2C的高表达样本富集于蛋白质分泌、有丝分裂纺锤体、PI3K/AKT/mTOR信号通路和mTORC1信号通路(FDR<0.1,|NES|>1,P<0.05)。KEGG信号通路分析显示:TFAP2C与相关基因主要通路富集于ErbB信号通路、EGFR酪氨酸激酶抑制剂耐药性和膀胱癌等(P<0.05)。GO功能富集分析显示:TFAP2C与相关基因的生物学过程主要富集于转录共激活因子活性、转录辅助调节因子活性和表皮生长因子受体结合等(P<0.05)。结论:TFAP2C在膀胱癌中起促癌作用,表达水平上调和预后不良有关,TFAP2C可能是判断膀胱癌预后的生物标志物。
基金supported by the National Natural Science Foundation of China(Nos.32230057,81970747,32271193)the National Key Research and Development Program of China(No.2017YFA0701304)the Priority Academic Program Development(PAPD)of Jiangsu Higher Education Institutions.
文摘Chitosan and its degradation product,oligosaccharides,have been shown to facilitate peripheral nerve regeneration.However,the underlying mechanisms are not well understood.In this study,we analyzed the protein expression profiles in sciatic nerves after injury using proteomics.A group of proteins related to exosome packaging and transport is up-regulated by chitosan oligosaccharides(COS),implying that exosomes are involved in COS-induced peripheral nerve regeneration.In fact,exosomes derived from fibroblasts(f-EXOs)treated with COS significantly promoted axon extension and regeneration.Exosomal protein identification and functional studies,revealed that TFAP2C is a key factor in neurite outgrowth induced by COS-f-EXOs.Furthermore,we showed that TFAP2C targets the pri-miRNA-132 gene and represses miR-132-5p expression in dorsal root ganglion neurons.Camkk1 is a downstream substrate of miR-132-5p that positively affects axon extension.In rats,miR-132-5p antagomir stimulates CAMKK1 expression and improves axon regeneration and functional recovery in sciatic nerves after injury.Our data reveal the mechanism for COS in axon regeneration,that is COS induce fibroblasts to produce TFAP2C-enriched EXOs,which are then transferred into axons to promote axon regeneration via miR-132-5p/CAMKK1.Moreover,these results show a new facet of fibroblasts in axon regeneration in peripheral nerves.
