Objective: This study was to investigate the role of hsa-miR-155-3p and hsa-miR-155-5p as biomarkers and regulators of biological behavior in Systemic Sclerosis. Methods: A total of 10 SSc patients and 10 healthy cont...Objective: This study was to investigate the role of hsa-miR-155-3p and hsa-miR-155-5p as biomarkers and regulators of biological behavior in Systemic Sclerosis. Methods: A total of 10 SSc patients and 10 healthy controls were selected for the study. The expression levels of hsa-miR-155-3p and hsa-miR-155-5p in peripheral blood mononuclear cells of SSc patients and healthy controls were measured using RT-qPCR. The diagnostic value of these miRNAs was explored using Receiver Operating Characteristic curve analysis. Pearson or Spearman correlation analysis was performed to assess the correlation between miRNAs and clinical indicators in SSc patients. Potential target genes of hsa-miR-155-3p and hsa-miR-155-5p were predicted using miRDB, Targetscan, and miRDIP databases. GO functional annotation, KEGG pathway enrichment analysis, protein-protein interaction network construction, and selection of central genes were conducted. Results: The expression levels of hsa-miR-155-3p and hsa- miR-155-5p were significantly higher in PBMCs of SSc patients compared to healthy controls (P<0.001). The ROC curve analysis showed that hsa-miR-155-3p and hsa-miR-155-5p had a high diagnostic value for SSc (AUC=1, P<0.001). Correlation analysis revealed that hsa- miR-155-3p, hsa-miR-155-5p, and clinical indicators such as high-resolution CT, neutrophil percentage, lymphocyte percentage, and albumin to globulin ratio were correlated (P<0.05). The signaling pathways enriched with target genes of hsa-miR-155-3p and hsa-miR-155- 5p were closely associated with the occurrence and development of SSc fibrosis, immunity, and inflammation. Conclusions: hsa-miR-155-3p and hsa-miR-155-5p may be involved in regulating the occurrence and development of SSc fibrosis, immunity, and inflammation. They have the potential to serve as biomarkers for clinical diagnosis and treatment of SSc.展开更多
Objective To investigate the role and molecular mechanism of exosomal miR-224-5p in colorectal cancer(CRC).Methods The miR-224-5p expression in CRC patient tissues and cell-derived exosomes was measured by laser captu...Objective To investigate the role and molecular mechanism of exosomal miR-224-5p in colorectal cancer(CRC).Methods The miR-224-5p expression in CRC patient tissues and cell-derived exosomes was measured by laser capture microdissection and qRT-PCR,respectively.Dual-luciferase reporter gene assay was used to determine the target gene of miR-224-5p.The protein expressions of p53 and unc-51 like kinase 2(ULK2)in CRC cells were detected by western blot.Flow cytometry was used to detect cell cycle and apoptosis.Cell proliferation was measured by CCK8 and EdU assay.Results The miR-224-5p expression was upregulated in CRC tissues and increased progressively with the rise of CRC stage.CRC cells secreted extracellular miR-224-5p mainly in an exosome-dependent manner,and then miR-224-5p could be transferred to surrounding tumor cells to regulate cell proliferation in the form of autocrine or paracrine.Moreover,ULK2 was characterized as a direct target of miR-224-5p and was downregulated in CRC tissues.Interestingly,ULK2 inhibited CRC cell proliferation in a p53-dependent manner.Furthermore,exosome-derived miR-224-5p partially reversed the proliferation regulation of ULK2 on CRC cells.Conclusion Our findings demonstrate that exosome-transmitted miR-224-5p promotes p53-dependent cell proliferation by targeting ULK2 in CRC,which may offer promising targets for CRC prevention and therapy.展开更多
背景与目的肺腺癌(lung adenocarcinoma,LUAD)是肺癌的一种主要亚型,其治疗与诊断依然是目前的研究热点。靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein 2,TPX2)在多种癌细胞中高表达,可能与LUAD的发生发展相关...背景与目的肺腺癌(lung adenocarcinoma,LUAD)是肺癌的一种主要亚型,其治疗与诊断依然是目前的研究热点。靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein 2,TPX2)在多种癌细胞中高表达,可能与LUAD的发生发展相关。本研究旨在探究TPX2对LUAD细胞恶性进程的影响以及调控机制。方法通过生物信息学分析技术,对癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中LUAD组织中基因TPX2的表达情况进行分析。实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测人肺正常细胞系和人LUAD细胞系中TPX2和miR-218-5p的表达水平。蛋白质印迹法(Western blot)检测细胞系中TPX2蛋白表达以及其对p53信号通路关键蛋白表达的影响。使用生物信息学预测并通过双荧光素酶报告基因检测验证TPX2与miR-218-5p的关系,细胞活力检测(cell counting kit-8,CCK-8)、细胞克隆形成、细胞划痕、Transwell实验、流式细胞术检测miR-218-5p和TPX2对LUAD细胞功能的影响。结果LUAD细胞中TPX2显著高表达,敲低TPX2可以抑制LUAD细胞的增殖、迁移、侵袭,促进凋亡并产生G_(2)/M期阻滞,并且促进p53信号通路关键蛋白的表达。miR-218-5p是TPX2的上游调控因子,可以抑制其表达。过表达miR-218-5p能消除TPX2高表达导致的恶性发展,抑制LUAD细胞的增殖、迁移等恶性进程以及促进p53信号通路。结论miR-218-5p靶向抑制TPX2表达,并通过p53发挥抑制LUAD细胞恶性发展的作用。展开更多
基金National Natural Science Foundation of China(No.8186029481860295)Natural Science Foundation of Inner Mongolia Autonomous Region(No.2019MS080552021MS08045)Science and Technology Plan Project of Inner Mongolia Autonomous Region(No.2018020892019GG052)。
文摘Objective: This study was to investigate the role of hsa-miR-155-3p and hsa-miR-155-5p as biomarkers and regulators of biological behavior in Systemic Sclerosis. Methods: A total of 10 SSc patients and 10 healthy controls were selected for the study. The expression levels of hsa-miR-155-3p and hsa-miR-155-5p in peripheral blood mononuclear cells of SSc patients and healthy controls were measured using RT-qPCR. The diagnostic value of these miRNAs was explored using Receiver Operating Characteristic curve analysis. Pearson or Spearman correlation analysis was performed to assess the correlation between miRNAs and clinical indicators in SSc patients. Potential target genes of hsa-miR-155-3p and hsa-miR-155-5p were predicted using miRDB, Targetscan, and miRDIP databases. GO functional annotation, KEGG pathway enrichment analysis, protein-protein interaction network construction, and selection of central genes were conducted. Results: The expression levels of hsa-miR-155-3p and hsa- miR-155-5p were significantly higher in PBMCs of SSc patients compared to healthy controls (P<0.001). The ROC curve analysis showed that hsa-miR-155-3p and hsa-miR-155-5p had a high diagnostic value for SSc (AUC=1, P<0.001). Correlation analysis revealed that hsa- miR-155-3p, hsa-miR-155-5p, and clinical indicators such as high-resolution CT, neutrophil percentage, lymphocyte percentage, and albumin to globulin ratio were correlated (P<0.05). The signaling pathways enriched with target genes of hsa-miR-155-3p and hsa-miR-155- 5p were closely associated with the occurrence and development of SSc fibrosis, immunity, and inflammation. Conclusions: hsa-miR-155-3p and hsa-miR-155-5p may be involved in regulating the occurrence and development of SSc fibrosis, immunity, and inflammation. They have the potential to serve as biomarkers for clinical diagnosis and treatment of SSc.
基金supported by the National Natural Science Foundation of China[Grant Number:81972803]。
文摘Objective To investigate the role and molecular mechanism of exosomal miR-224-5p in colorectal cancer(CRC).Methods The miR-224-5p expression in CRC patient tissues and cell-derived exosomes was measured by laser capture microdissection and qRT-PCR,respectively.Dual-luciferase reporter gene assay was used to determine the target gene of miR-224-5p.The protein expressions of p53 and unc-51 like kinase 2(ULK2)in CRC cells were detected by western blot.Flow cytometry was used to detect cell cycle and apoptosis.Cell proliferation was measured by CCK8 and EdU assay.Results The miR-224-5p expression was upregulated in CRC tissues and increased progressively with the rise of CRC stage.CRC cells secreted extracellular miR-224-5p mainly in an exosome-dependent manner,and then miR-224-5p could be transferred to surrounding tumor cells to regulate cell proliferation in the form of autocrine or paracrine.Moreover,ULK2 was characterized as a direct target of miR-224-5p and was downregulated in CRC tissues.Interestingly,ULK2 inhibited CRC cell proliferation in a p53-dependent manner.Furthermore,exosome-derived miR-224-5p partially reversed the proliferation regulation of ULK2 on CRC cells.Conclusion Our findings demonstrate that exosome-transmitted miR-224-5p promotes p53-dependent cell proliferation by targeting ULK2 in CRC,which may offer promising targets for CRC prevention and therapy.
文摘背景与目的肺腺癌(lung adenocarcinoma,LUAD)是肺癌的一种主要亚型,其治疗与诊断依然是目前的研究热点。靶向Xklp2靶蛋白(targeting protein for Xenopus kinesin-like protein 2,TPX2)在多种癌细胞中高表达,可能与LUAD的发生发展相关。本研究旨在探究TPX2对LUAD细胞恶性进程的影响以及调控机制。方法通过生物信息学分析技术,对癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中LUAD组织中基因TPX2的表达情况进行分析。实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测人肺正常细胞系和人LUAD细胞系中TPX2和miR-218-5p的表达水平。蛋白质印迹法(Western blot)检测细胞系中TPX2蛋白表达以及其对p53信号通路关键蛋白表达的影响。使用生物信息学预测并通过双荧光素酶报告基因检测验证TPX2与miR-218-5p的关系,细胞活力检测(cell counting kit-8,CCK-8)、细胞克隆形成、细胞划痕、Transwell实验、流式细胞术检测miR-218-5p和TPX2对LUAD细胞功能的影响。结果LUAD细胞中TPX2显著高表达,敲低TPX2可以抑制LUAD细胞的增殖、迁移、侵袭,促进凋亡并产生G_(2)/M期阻滞,并且促进p53信号通路关键蛋白的表达。miR-218-5p是TPX2的上游调控因子,可以抑制其表达。过表达miR-218-5p能消除TPX2高表达导致的恶性发展,抑制LUAD细胞的增殖、迁移等恶性进程以及促进p53信号通路。结论miR-218-5p靶向抑制TPX2表达,并通过p53发挥抑制LUAD细胞恶性发展的作用。
