目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜...目的:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜癌细胞株Ishikawa转染si-TFAP2A-AS1(si-TFAP2A-AS1组)、si-NC(si-NC组)、miR-9-5p mimics(miR-9-5p组)及miR-NC(miR-NC组);RL95-2细胞转染OE-TFAP2A-AS1(TFAP2A-AS1组)、Vector(Vector组)、sh-miR-9-5p(sh-miR-9-5p组)及sh-NC(sh-NC组)。CCK-8检测细胞增殖能力,Transwell检测细胞侵袭、迁移能力,双荧光素酶实验、pull down实验分析TFAP2A-AS1与miR-9-5p的靶向关系;miR-9-5p与ERK的靶向关系,实时荧光定量PCR(RT-qPCR)检测子宫内膜癌组织、癌旁组织、子宫内膜癌细胞及hEEC细胞TFAP2AAS1、miR-9-5p表达水平,Western blot检测细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)水平。结果:子宫内膜癌组织中TFAP2A-AS1表达水平低于癌旁组织,miR-9-5p表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织TFAP2A-AS1、miR-9-5p表达水平与分化程度、肿瘤直径、TNM分期及淋巴结转移有关。子宫内膜癌组织中TFAP2A-AS1、miR-9-5p表达水平呈负相关(r=-0.782,P=0.002)。与hEEC细胞比较,RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa细胞TFAP2A-AS1表达水平降低,miR-9-5p表达水平升高(P<0.05)。与si-NC组比较,si-TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与Vector组比较,TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。与miR-NC组比较,miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与sh-NC组比较,sh-miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。TFAP2A-AS1负性调控miR-9-5p、p-ERK表达水平,miR-9-5p可靶向调控ERK蛋白。结论:过表达TFAP2A-AS1通过靶向下调miR-9-5p,抑制ERK通路,最终抑制子宫内膜癌细胞恶性生物学行为。展开更多
In this study,we mainly focus on probing expression profile and detailed functions of long non-coding RNA TFAP2A antisense RNA 1(TFAP2A-AS1)in non-small cell lung cancer(NSCLC).Moreover,the mechanisms played by TFAP2A...In this study,we mainly focus on probing expression profile and detailed functions of long non-coding RNA TFAP2A antisense RNA 1(TFAP2A-AS1)in non-small cell lung cancer(NSCLC).Moreover,the mechanisms played by TFAP2A-AS1 were unraveled comprehensively.Herein,a notable overexpressed TFAP2A-AS1 in NSCLC was observed by TCGA and our own cohort.An increased TFAP2A-AS1 level displayed a negative correlation with the overall survival of patients with NSCLC.Loss-of-function approaches illustrated that the absence of TFAP2A-AS1 weakened NSCLC cell proliferation,colony formation,migration and invasion in vitro.Also,interference of TFAP2A-AS1 caused in vivo tumor growth suppression.Mechanistically,TFAP2A-AS1 could negative regulate microRNA-584-3p(miR-584-3p)as a competitive endogenous RNA.Furthermore,cyclin-dependent kinase 4(CDK4),a direct target of miR-584-3p,was positively controlled by TFAP2A-AS1 in a miR-5184-3p-dependent manner.Rescue function experiments corroborated that the anticancer activities of TFAP2A-AS1 deficient on the oncogenicity of NSCLC cells were reversed by downregulating miR-584-3p or overexpressing CDK4.To sum up,TFAP2A-AS1 exhibits cancerpromoting roles in NSCLC through the adjustment of miR-584-3p/CDK4 axis.展开更多
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:探讨长链非编码RNA TFAP2A-AS1对子宫内膜癌细胞增殖、侵袭和迁移的影响及机制。方法:选择50例子宫内膜癌组织和对应的癌旁组织。选取子宫内膜癌细胞株(RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa),子宫内膜上皮细胞hEEC。子宫内膜癌细胞株Ishikawa转染si-TFAP2A-AS1(si-TFAP2A-AS1组)、si-NC(si-NC组)、miR-9-5p mimics(miR-9-5p组)及miR-NC(miR-NC组);RL95-2细胞转染OE-TFAP2A-AS1(TFAP2A-AS1组)、Vector(Vector组)、sh-miR-9-5p(sh-miR-9-5p组)及sh-NC(sh-NC组)。CCK-8检测细胞增殖能力,Transwell检测细胞侵袭、迁移能力,双荧光素酶实验、pull down实验分析TFAP2A-AS1与miR-9-5p的靶向关系;miR-9-5p与ERK的靶向关系,实时荧光定量PCR(RT-qPCR)检测子宫内膜癌组织、癌旁组织、子宫内膜癌细胞及hEEC细胞TFAP2AAS1、miR-9-5p表达水平,Western blot检测细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)水平。结果:子宫内膜癌组织中TFAP2A-AS1表达水平低于癌旁组织,miR-9-5p表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织TFAP2A-AS1、miR-9-5p表达水平与分化程度、肿瘤直径、TNM分期及淋巴结转移有关。子宫内膜癌组织中TFAP2A-AS1、miR-9-5p表达水平呈负相关(r=-0.782,P=0.002)。与hEEC细胞比较,RL95-2、HEC-1A、HHUA、HEC-1B及Ishikawa细胞TFAP2A-AS1表达水平降低,miR-9-5p表达水平升高(P<0.05)。与si-NC组比较,si-TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与Vector组比较,TFAP2A-AS1组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。与miR-NC组比较,miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均升高(P<0.05);与sh-NC组比较,sh-miR-9-5p组细胞48 h、72 h OD值,侵袭、迁移细胞数均降低(P<0.05)。TFAP2A-AS1负性调控miR-9-5p、p-ERK表达水平,miR-9-5p可靶向调控ERK蛋白。结论:过表达TFAP2A-AS1通过靶向下调miR-9-5p,抑制ERK通路,最终抑制子宫内膜癌细胞恶性生物学行为。
文摘In this study,we mainly focus on probing expression profile and detailed functions of long non-coding RNA TFAP2A antisense RNA 1(TFAP2A-AS1)in non-small cell lung cancer(NSCLC).Moreover,the mechanisms played by TFAP2A-AS1 were unraveled comprehensively.Herein,a notable overexpressed TFAP2A-AS1 in NSCLC was observed by TCGA and our own cohort.An increased TFAP2A-AS1 level displayed a negative correlation with the overall survival of patients with NSCLC.Loss-of-function approaches illustrated that the absence of TFAP2A-AS1 weakened NSCLC cell proliferation,colony formation,migration and invasion in vitro.Also,interference of TFAP2A-AS1 caused in vivo tumor growth suppression.Mechanistically,TFAP2A-AS1 could negative regulate microRNA-584-3p(miR-584-3p)as a competitive endogenous RNA.Furthermore,cyclin-dependent kinase 4(CDK4),a direct target of miR-584-3p,was positively controlled by TFAP2A-AS1 in a miR-5184-3p-dependent manner.Rescue function experiments corroborated that the anticancer activities of TFAP2A-AS1 deficient on the oncogenicity of NSCLC cells were reversed by downregulating miR-584-3p or overexpressing CDK4.To sum up,TFAP2A-AS1 exhibits cancerpromoting roles in NSCLC through the adjustment of miR-584-3p/CDK4 axis.
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.
