5种尿液修饰核苷联合血浆EBV-DNA定量检测在鼻咽癌诊断中的可行性研究

Feasibility study of combined detection of five urinary modified nucleosides and plasma EBV-DNA in the diagnosis of nasopharyngeal carcinoma

目的探讨5种尿液修饰核苷联合血浆EBV-DNA定量检测在鼻咽癌诊断中的可行性。方法选取2020年1~6月浙江省肿瘤医院收治的56例初诊鼻咽癌患者作为鼻咽癌组,选取同期32例健康体检者作为正常对照组。采用液相色谱-串联质谱方法检测尿液中修饰核苷假尿嘧啶核苷(Pseu)、1-甲基腺苷(m1A)、2-甲基鸟苷(m2G)、6-甲基腺苷(m6A)和1-甲基鸟苷(m1G)的浓度,比较两组受试者尿液修饰核苷的表达水平。采用荧光定量聚合酶链反应技术检测血浆EBV-DNA的拷贝数,并制作受试者工作特征(ROC)曲线评价尿液核苷和血浆EBV-DNA单项检测及联合检测的诊断价值。结果鼻咽癌组尿液中的m2G、Pseu、m1A、m1G浓度均高于正常对照组,差异有统计学意义(P<0.05)。鼻咽癌组的血浆EBV-DNA拷贝数高于正常对照组,差异有统计学意义(P<0.05)。m2G、m1A、Pseu、m1G与EBV-DNA 5项联合检测的曲线下面积(AUC)均高于m2G、m1A、Pseu、m1G与EBV-DNA单项检测,差异有统计学意义(P<0.05)。Pseu浓度与临床分期及淋巴结转移均呈正相关(r=0.474、0.406,P<0.01)。结论尿液Pseu、m2G、m1A和m1G有望成为诊断鼻咽癌的小分子标志物,上述4种尿液核苷与血浆EBV-DNA 5项联合检测可能有助于提高诊断鼻咽癌的诊断效能。

Objective To explore the feasibility of combined detection of five urinary modified nucleosides and plasma EBV-DNA in the diagnosis of nasopharyngeal carcinoma.Methods A total of 56 patients with nasopharyngeal carcinoma treated in Zhejiang Cancer Hospital from January to June 2020 were selected as the nasopharyngeal carcinoma group and 32 healthy subjects during the same period were selected as the healthy control group.Liquid chromatography-tandem mass spectrometry was employed to measure the concentrations of modified nucleosides pseudouridine(Pseu),2-O-methylguanosine(m2G),1-methyladenosine(m1A),6-methyladenosine(m6A)and 1-methylguanosine(m1G)in urine samples.The levels of Pseu,m2G,m1A,m6A and m1G in urine samples of the two groups were compared.Fluorescent quantitative polymerase chain reaction was employed to measure the level of the plasma EBV-DNA in peripheral blood samples collected from all the subjects.Receiver operating characteristic(ROC)curve was made to evaluate the diagnostic value of single detection and combined detection of urinary nucleosides and plasma EBV-DNA.Results The levels of Pseu,m2G,m1A and m1G in the nasopharyngeal carcinoma group were higher than those in the healthy control group,the differences were statistically significant(P<0.05).The level of the plasma EBV-DNA in the nasopharyngeal carcinoma group was higher than that in the healthy control group,the difference was statistically significant(P<0.05).The area under the curve(AUC)of the combined detection was higher than that of m2G,m1A,Pseu,m1G and EBV-DNA in individuals,the differences were statistically significant(P<0.05).In the nasopharyngeal carcinoma patients,the Pseu concentration was positively correlated with clinical stage and lymph node metastasis(r=0.474,0.406,P<0.01).Conclusion The urinary modified nucleosides Pseu,m2G,m1A and m1G may be small molecular markers for the diagnosis of nasopharyngeal carcinoma.The combined detection of the four urinary nucleosides with plasma EBV-DNA may help to improve the diagnostic efficiency of nasopharyngeal carcinoma.