Regulation of microRNAs(miRNAs)on various biological processes has been a surprising and exciting field.Identification of miRNAs is the first step to comprehensively understand their functions.However,attempts on glob...Regulation of microRNAs(miRNAs)on various biological processes has been a surprising and exciting field.Identification of miRNAs is the first step to comprehensively understand their functions.However,attempts on global identification and functional verification of miRNAs are very limited in penaeid shrimp Marsupenaeus japonicus,an economically important aquatic species.By performing an integrated analysis of transcriptomic profile from gastrula embryos of M.japonicus,21conserved miRNAs in M.japonicas(mja-miRNAs),belonging to 19 miRNA families,were identified and characterized.Of the 21 mja-miRNAs,15 miRNAs were successfully verified to be predominantly expressed in gastrula stage,where they displayed dynamic expression patterns compared with those in naupliuin stage.Based on perfect or near-perfect match to target region,120 target genes were predicted at transcriptome-wide level.Noteworthy,gene ontology(GO)classification and metabolic pathway annotation revealed eight targets that were actively involved in developmental processes.Of the predicted miRNA-mRNA pairs,six targets were then randomly selected and experimentally validated by dual luciferase reporter assay,where three pairs were proved with potential targeting activity.Overall,to search for conserved miRNAs potentially involved in early development of M.japonicus,we combined in silico and experimental methods,which can be applied in other organisms as well.Our data implied important roles of miRNAs in the early embryonic development and also suggested the presence of complex miRNA-mRNA functional networks in M.japonicus.展开更多
目的探索胃癌发生过程的风险miRNAs,为上消化道机会性筛查中早期胃癌识别提供依据。方法纳入2021年6月至2023年8月在广西医科大学附属肿瘤医院、右江民族医学院附属医院、桂林市人民医院3个中心进行上消化道癌机会性筛查的人群。选取健...目的探索胃癌发生过程的风险miRNAs,为上消化道机会性筛查中早期胃癌识别提供依据。方法纳入2021年6月至2023年8月在广西医科大学附属肿瘤医院、右江民族医学院附属医院、桂林市人民医院3个中心进行上消化道癌机会性筛查的人群。选取健康体检者107例、早期胃癌患者71例、进展期胃癌患者97例。首先采用转录组测序筛选差异表达miRNAs,然后在3组前瞻性人群的血浆样本中通过RT‐qPCR验证差异表达的miRNAs,最后采用受试者工作特征(receiver operating characteristic,ROC)曲线评估miRNAs的诊断效能。结果转录组测序的差异基因分析共筛选出胃癌发生过程中的6个差异表达miRNAs,包括miR‐3176、miR‐885‐5p、miR‐203a‐3p、miR‐452‐5p、miR‐223‐3p、miR‐219a‐2‐3p。RT‐qPCR结果显示,miR‐452‐5p在早期胃癌及进展期胃癌患者中表达上调(均P<0.001)。ROC曲线显示,miR‐452‐5p诊断早期胃癌和进展期胃癌的曲线下面积(area under the curve,AUC)分别为0.900、0.975。区分早期胃癌与进展期胃癌的AUC为0.843。结论miR‐452‐5p在早期胃癌中具有良好的诊断效能,可能作为液体活检诊断早期胃癌的潜在生物标志物。展开更多
基金the National Natural Science Foundation of China(No.32273116)the Natural Science Foundation of Shandong Province(China)(No.ZR2020MC189)the National Key R&D Program of China(No.2018YFD0901301)。
文摘Regulation of microRNAs(miRNAs)on various biological processes has been a surprising and exciting field.Identification of miRNAs is the first step to comprehensively understand their functions.However,attempts on global identification and functional verification of miRNAs are very limited in penaeid shrimp Marsupenaeus japonicus,an economically important aquatic species.By performing an integrated analysis of transcriptomic profile from gastrula embryos of M.japonicus,21conserved miRNAs in M.japonicas(mja-miRNAs),belonging to 19 miRNA families,were identified and characterized.Of the 21 mja-miRNAs,15 miRNAs were successfully verified to be predominantly expressed in gastrula stage,where they displayed dynamic expression patterns compared with those in naupliuin stage.Based on perfect or near-perfect match to target region,120 target genes were predicted at transcriptome-wide level.Noteworthy,gene ontology(GO)classification and metabolic pathway annotation revealed eight targets that were actively involved in developmental processes.Of the predicted miRNA-mRNA pairs,six targets were then randomly selected and experimentally validated by dual luciferase reporter assay,where three pairs were proved with potential targeting activity.Overall,to search for conserved miRNAs potentially involved in early development of M.japonicus,we combined in silico and experimental methods,which can be applied in other organisms as well.Our data implied important roles of miRNAs in the early embryonic development and also suggested the presence of complex miRNA-mRNA functional networks in M.japonicus.
文摘目的探索胃癌发生过程的风险miRNAs,为上消化道机会性筛查中早期胃癌识别提供依据。方法纳入2021年6月至2023年8月在广西医科大学附属肿瘤医院、右江民族医学院附属医院、桂林市人民医院3个中心进行上消化道癌机会性筛查的人群。选取健康体检者107例、早期胃癌患者71例、进展期胃癌患者97例。首先采用转录组测序筛选差异表达miRNAs,然后在3组前瞻性人群的血浆样本中通过RT‐qPCR验证差异表达的miRNAs,最后采用受试者工作特征(receiver operating characteristic,ROC)曲线评估miRNAs的诊断效能。结果转录组测序的差异基因分析共筛选出胃癌发生过程中的6个差异表达miRNAs,包括miR‐3176、miR‐885‐5p、miR‐203a‐3p、miR‐452‐5p、miR‐223‐3p、miR‐219a‐2‐3p。RT‐qPCR结果显示,miR‐452‐5p在早期胃癌及进展期胃癌患者中表达上调(均P<0.001)。ROC曲线显示,miR‐452‐5p诊断早期胃癌和进展期胃癌的曲线下面积(area under the curve,AUC)分别为0.900、0.975。区分早期胃癌与进展期胃癌的AUC为0.843。结论miR‐452‐5p在早期胃癌中具有良好的诊断效能,可能作为液体活检诊断早期胃癌的潜在生物标志物。
