The mechanisms that regulate neural stem cell(NSC)lineage progression and maintain NSCs within diffe rent domains of the adult neural stem cell niche,the subventricular zone are not well defined.Quiescent NSCs are arr...The mechanisms that regulate neural stem cell(NSC)lineage progression and maintain NSCs within diffe rent domains of the adult neural stem cell niche,the subventricular zone are not well defined.Quiescent NSCs are arranged at the apical ventricular wall,while mitotically activated NSCs are found in the basal,vascular region of the subventricular zone.Here,we found that ADAM 10(a disintegrin and metalloproteinase 10)is essential in NSC association with the ventricular wall,and via this adhesion to the apical domain,ADAM10 regulates the switch from quiescent and undiffe rentiated NSC to an actively prolife rative and differentiating cell state.Processing of JAMC(junctional adhesion molecule C)by ADAM 10 increases Rap1 GAP activity.This molecular machinery promotes NSC transit from the apical to the basal compartment and subsequent lineage progression.Understanding the molecular mechanisms responsible for regulating the proper positioning of NSCs within the subventricular zone niche and lineage progression of NSCs could provide new targets for drug development to enhance the regenerative prope rties of neural tissue.展开更多
目的探讨实时荧光定量PCR SYBR Green I探针技术在研究肝癌患者中基因表达情况的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测肝癌患者癌组织及癌旁组织RAP1GAP基因表达情况。结果实验组和对照组有明显的统计学差异。结论Rea...目的探讨实时荧光定量PCR SYBR Green I探针技术在研究肝癌患者中基因表达情况的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测肝癌患者癌组织及癌旁组织RAP1GAP基因表达情况。结果实验组和对照组有明显的统计学差异。结论Real-time PCR技术检测目的基因的表达是非常方便快捷准确的方法。展开更多
Background:Although Mex3 RNA-binding family member A(Mex3a)has demonstrated an important role in multiple cancers,its role and regulatory mechanism in CRC is unclear.In this study,we aimed to investigate the role and ...Background:Although Mex3 RNA-binding family member A(Mex3a)has demonstrated an important role in multiple cancers,its role and regulatory mechanism in CRC is unclear.In this study,we aimed to investigate the role and clinical significance of Mex3a in CRC and to explore its underlying mechanism.Methods:Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were performed to detect the expression levels of genes.5-Ethynyl-2’-deoxyuridine(EDU)and transwell assays were utilized to examine CRC cell proliferation and metastatic ability.The R software was used to do hierarchical clustering analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Overexpression and rescue experiments which included U0126,a specific mitogen activated protein kinase kinase/extracellular regulated protein kinase(MEK/ERK)inhibitor,and PX-478,a hypoxia-inducible factor 1 subunit alpha(HIF-1α)inhibitor,were used to study the molecularmechanisms of Mex3a in CRC cells.Co-immunoprecipitation(Co-IP)assay was performed to detect the interaction between two proteins.Bioinformatics analysis including available public database and Starbase software(starbase.sysu.edu.cn)were used to evaluate the expression and prognostic significance of genes.TargetScan(www.targetscan.org)and the miRDB(mirdb.org)website were used to predict the combination site between microRNA and target mRNA.BALB/c nude micewere used to study the function of Mex3a and hsa-miR-6887-3p in vivo.Results:Clinicopathological and immunohistochemical(IHC)studies of 101 CRC tissues and 79 normal tissues demonstrated that Mex3a was a significant prognostic factor for overall survival(OS)in CRC patients.Mex3a knockdown substantially inhibited the migration,invasion,and proliferation of CRC cells.Transcriptome analysis and mechanism verification showed that Mex3a regulated the RAP1 GTPase activating protein(RAP1GAP)/MEK/ERK/HIF-1αpathway.Furthermore,RAP1GAP was identified to interact with Mex3a in Co-IP experiments.Bioinformatics and dual-luciferase reporter experiments revealed that hsa-miR-6887-3p could bind to the 3’-untranslated regions(3’-UTR)of the Mex3amRNA.hsa-miR-6887-3p downregulated Mex3a expression and inhibited the tumorigenesis of CRC both in vitro and in vivo.Conclusions:Our study demonstrated that the hsa-miR-6887-3p/Mex3a/RAP1GAP signaling axis was a key regulator of CRC and Mex3a has the potential to be a new diagnostic marker and treatment target for CRC.展开更多
基金National Institutes of Health(NIH)Grants R01 RMH099384(to AA)and T32GM008444(to NM)。
文摘The mechanisms that regulate neural stem cell(NSC)lineage progression and maintain NSCs within diffe rent domains of the adult neural stem cell niche,the subventricular zone are not well defined.Quiescent NSCs are arranged at the apical ventricular wall,while mitotically activated NSCs are found in the basal,vascular region of the subventricular zone.Here,we found that ADAM 10(a disintegrin and metalloproteinase 10)is essential in NSC association with the ventricular wall,and via this adhesion to the apical domain,ADAM10 regulates the switch from quiescent and undiffe rentiated NSC to an actively prolife rative and differentiating cell state.Processing of JAMC(junctional adhesion molecule C)by ADAM 10 increases Rap1 GAP activity.This molecular machinery promotes NSC transit from the apical to the basal compartment and subsequent lineage progression.Understanding the molecular mechanisms responsible for regulating the proper positioning of NSCs within the subventricular zone niche and lineage progression of NSCs could provide new targets for drug development to enhance the regenerative prope rties of neural tissue.
文摘目的探讨实时荧光定量PCR SYBR Green I探针技术在研究肝癌患者中基因表达情况的可行性。方法运用实时荧光定量PCR SYBR Green I技术,检测肝癌患者癌组织及癌旁组织RAP1GAP基因表达情况。结果实验组和对照组有明显的统计学差异。结论Real-time PCR技术检测目的基因的表达是非常方便快捷准确的方法。
基金supported by the National Scientific Foundation of China(NSFC32071127 and 31871160)。
文摘Background:Although Mex3 RNA-binding family member A(Mex3a)has demonstrated an important role in multiple cancers,its role and regulatory mechanism in CRC is unclear.In this study,we aimed to investigate the role and clinical significance of Mex3a in CRC and to explore its underlying mechanism.Methods:Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were performed to detect the expression levels of genes.5-Ethynyl-2’-deoxyuridine(EDU)and transwell assays were utilized to examine CRC cell proliferation and metastatic ability.The R software was used to do hierarchical clustering analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Overexpression and rescue experiments which included U0126,a specific mitogen activated protein kinase kinase/extracellular regulated protein kinase(MEK/ERK)inhibitor,and PX-478,a hypoxia-inducible factor 1 subunit alpha(HIF-1α)inhibitor,were used to study the molecularmechanisms of Mex3a in CRC cells.Co-immunoprecipitation(Co-IP)assay was performed to detect the interaction between two proteins.Bioinformatics analysis including available public database and Starbase software(starbase.sysu.edu.cn)were used to evaluate the expression and prognostic significance of genes.TargetScan(www.targetscan.org)and the miRDB(mirdb.org)website were used to predict the combination site between microRNA and target mRNA.BALB/c nude micewere used to study the function of Mex3a and hsa-miR-6887-3p in vivo.Results:Clinicopathological and immunohistochemical(IHC)studies of 101 CRC tissues and 79 normal tissues demonstrated that Mex3a was a significant prognostic factor for overall survival(OS)in CRC patients.Mex3a knockdown substantially inhibited the migration,invasion,and proliferation of CRC cells.Transcriptome analysis and mechanism verification showed that Mex3a regulated the RAP1 GTPase activating protein(RAP1GAP)/MEK/ERK/HIF-1αpathway.Furthermore,RAP1GAP was identified to interact with Mex3a in Co-IP experiments.Bioinformatics and dual-luciferase reporter experiments revealed that hsa-miR-6887-3p could bind to the 3’-untranslated regions(3’-UTR)of the Mex3amRNA.hsa-miR-6887-3p downregulated Mex3a expression and inhibited the tumorigenesis of CRC both in vitro and in vivo.Conclusions:Our study demonstrated that the hsa-miR-6887-3p/Mex3a/RAP1GAP signaling axis was a key regulator of CRC and Mex3a has the potential to be a new diagnostic marker and treatment target for CRC.
