血浆外泌体miR-548k在食管鳞癌中的表达及诊断价值

Expression and diagnostic value of plasma exosomal miR-548k in esophageal squamous cell carcinoma

目的探讨血浆外泌体中与食管鳞癌(esophageal squamous cell carcinoma,ESCC)发生相关的microRNAs(miRNAs),并评估其在ESCC诊断中的应用价值。方法选取2020年3月至2021年6月山东大学第二医院胸外科收治的55例ESCC患者为ESCC组,另选取55例健康体检者为对照组,提取受试者血浆外泌体,并通过透射电镜、NanoSight粒径分析及Western blotting进行鉴定。筛选阶段,送检对照组、ESCC组术前血浆外泌体RNA各5例进行测序。采用qRT-PCR法对训练集、验证集中差异性miRNAs分子表达进行检测,并通过受试者工作特征(receiver operating characteristic,ROC)曲线评估差异性miRNAs分子的诊断价值。结果测序结果显示,与对照组相比,5个miRNAs(miR-548k、miR-6516-5p、let-7b-3p、miR-3934-5p、miR-196a-5p)在ESCC组血浆外泌体中表达显著上调,8个miRNAs(miR-34c-5p、novel_167、miR-548u、miR-548ah-3p、miR-548p、miR-200a-5p、miR-200b-3p、miR-1228-5p)明显下调。通过训练队列证实,miR-548k在ESCC组血浆外泌体中明显上调(P<0.001),血浆外泌体miR-548k用于ESCC诊断时ROC曲线下面积(area under curve,AUC)为0.918,明显优于miR-34c-5p、miR-548u、miR-6516-5p。通过验证队列进一步证实,miR-548k在ESCC组血浆外泌体中高表达(P<0.001),血浆外泌体miR-548k可用于ESCC患者诊断(AUC值为0.889)。结论miR-548k在ESCC组血浆外泌体中表达显著增高,血浆外泌体miR-548k对ESCC诊断有一定的临床应用价值。

Objective To screen out the candidate plasma exosomal microRNAs(miRNAs)which play crucial role in esophageal squamous cell carcinoma(ESCC)occurrence and evaluate their potential in ESCC diagnosis.Methods A total of 55 ESCC patients who underwent surgical resection at The Second Hospital of Shandong University from March 2020 to June 2021 and 55 healthy volunteers were enrolled in this study.After identified by transmission electron microscopy(TEM),NanoSight tracking analysis(NTA)and Western blotting assays,RNA sequencing was performed to determine the miRNA profiles of plasma exosomes from 5 ESCC patients and 5 healthy control.Then,quantitative real-time PCR(qRT-PCR)was used to detect the expression level of differential exosomal miRNAs in the training and validation sets.Then,receiver operating characteristic(ROC)curve was conducted to evaluate their diagnostic value.Results Five miRNAs(miR-548k,miR-6516-5p,let-7b-3p,miR-3934-5p,miR-196a-5p)were significantly higher,and 8 miRNAs(miR-34c-5p,novel_167,miR-548u,miR-548ah-3p,miR-548p,miR-200a-5p,miR-200b-3p,miR-1228-5p)were obviously decreased in plasma exosomes of ESCC group than that of control group.Additionally,through the training cohort,we verified that miR-548k was significantly up-regulated in plasma exosomes of ESCC group(P<0.001).ROC curve analysis revealed that exosomal miR-548k exhibited higher accuracy than miR-34c-5p,miR-548u,miR-6516-5p in ESCC diagnosis,and the area under ROC curve(AUC)value was 0.918.Finally,through the validation cohort,we further confirmed that miR-548k was highly expressed in plasma exosomes of ESCC group(P<0.001),and plasma exosomal miR-548k could be used for ESCC diagnosis(AUC=0.889).Conclusion MiR-548k is significantly up-regulated in plasma exosomes of ESCC patients and plasma exosomal miR-548k has high clinical diagnostic value for ESCC.