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
Objective: Significance of combined detection of serum 25(OH)D3, hsa-miR-17-5p and HCV-RNA in diagnosis of hepatitis C disease. Methods: A total of 52 patients with benign ovarian tumor resection were selected from Ap...Objective: Significance of combined detection of serum 25(OH)D3, hsa-miR-17-5p and HCV-RNA in diagnosis of hepatitis C disease. Methods: A total of 52 patients with benign ovarian tumor resection were selected from April 2016 to April 2018 in our hospital. We paid attention to the implementation of perioperative nursing intervention, and observed the prognosis and the occurrence of adverse reactions. Results: The expression of hsa-mir-17-5p in the observation group was significantly higher than that in the control group, the expression of 25(OH)D3 was lower than that in the control group, the expression of hsa-mir-17-5p in the HCV-RNA positive group was higher than that in the HCV-RNA negative group, and the expression of 25(OH) D3 was lower than that in the HCV-RNA negative group. The expression of HCV-RNA in the observation group was positively and negatively correlated with the expression of 25(OH) D3 and hsa-mir-17-5p in the 25(OH)D3 and the critical values of serum 25 (OH)D3, hsa-mir-17-5p and HCV-RNA were 24.23 ug/L, 1.89 relative expression and 1.22 × 103 IU/mL, respectively. The area of AUC detected by hsa-miR-17-5p+ 25 (OH) D3+25(OH)D3 is larger than that detected separately. Conclusion: The combined detection of serum 25(OH)D3, hsa-mir-17-5p and HCV-RNA has high diagnostic efficiency and sensitivity for hepatitis C. It is suggested that they be used in clinical practice.展开更多
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.
文摘目的·分析人类mi R-223-3p(hsa-miR-223-3p)的靶基因及其参与的生物学过程,并寻找与糖尿病相关的生物标志物。方法·利用starBase数据库筛选hsa-miR-223-3p靶基因,并对其行基因本体数据库(Gene Ontology,GO)富集分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genome,KEGG)和Reactome通路分析。通过构建蛋白-蛋白相互作用(proteinprotein interaction,PPI)网络获得中枢基因,筛选最有意义的模块,并利用Venn图对糖尿病相关的中枢基因进行分析。结果·共筛选出870个hsa-miR-223-3p靶基因。GO、KEGG和Reactome富集分析显示,靶基因主要与RNA聚合酶Ⅱ启动子的调节、细胞对胰岛素刺激的反应、RNA结合等相关,且主要富集于胰岛素分泌、泛素介导的蛋白水解、雌激素依赖型基因表达等通路。PPI网络共得31个中枢基因,且中枢基因PRKACB参与胰岛素分泌通路;共筛选出3个最有意义的基因模块,其中模块1参与泛素介导的蛋白水解作用,模块2参与RNA转运和细胞周期,模块3参与内吞作用。结论·Hsa-miR-223-3p可能通过靶基因参与多种生物学过程,中枢基因PRKACB或可为糖尿病发生机制的探索提供帮助。
文摘Objective: Significance of combined detection of serum 25(OH)D3, hsa-miR-17-5p and HCV-RNA in diagnosis of hepatitis C disease. Methods: A total of 52 patients with benign ovarian tumor resection were selected from April 2016 to April 2018 in our hospital. We paid attention to the implementation of perioperative nursing intervention, and observed the prognosis and the occurrence of adverse reactions. Results: The expression of hsa-mir-17-5p in the observation group was significantly higher than that in the control group, the expression of 25(OH)D3 was lower than that in the control group, the expression of hsa-mir-17-5p in the HCV-RNA positive group was higher than that in the HCV-RNA negative group, and the expression of 25(OH) D3 was lower than that in the HCV-RNA negative group. The expression of HCV-RNA in the observation group was positively and negatively correlated with the expression of 25(OH) D3 and hsa-mir-17-5p in the 25(OH)D3 and the critical values of serum 25 (OH)D3, hsa-mir-17-5p and HCV-RNA were 24.23 ug/L, 1.89 relative expression and 1.22 × 103 IU/mL, respectively. The area of AUC detected by hsa-miR-17-5p+ 25 (OH) D3+25(OH)D3 is larger than that detected separately. Conclusion: The combined detection of serum 25(OH)D3, hsa-mir-17-5p and HCV-RNA has high diagnostic efficiency and sensitivity for hepatitis C. It is suggested that they be used in clinical practice.
