ccfDNA中RERG甲基化状态检测在诊断鼻咽癌中的临床意义  

Clinical significance of RERG methylation status detection in circulating cell-free DNA for diagnosing nasopharyngeal carcinoma

作　　者：李宏丰 李刚[1] 云惟琳 张前飞 张秋叶 LI Hongfeng;LI Gang;YUN Weilin;ZHANG Qianfei;ZHANG Qiuye(Department of E.N.T.,Wenchang People′s Hospital,Wenchang,Hainan 571300,China;Department of Clinical Laboratory,Wenchang People′s Hospital,Wenchang,Hainan 571300,China)

机构地区：[1]文昌市人民医院耳鼻咽喉科,海南文昌571300 [2]文昌市人民医院检验科,海南文昌571300

出　　处：《国际检验医学杂志》2022年第9期1046-1050,1054,共6页International Journal of Laboratory Medicine

基　　金：海南省卫生计生行业科研项目(16A200006)。

摘　　要：目的研究循环细胞游离DNA(ccfDNA)中RAS样雌激素调节生长抑制因子(RERG)基因甲基化状态检测在诊断鼻咽癌中的临床意义。方法选取2020年1月至2021年6月在该院就诊的100例(病理学诊断)原发性鼻咽癌患者作为鼻咽癌组,同时选取97例非癌志愿者作为对照组。提取活检组织DNA和ccfDNA,通过实时荧光定量PCR法DNA甲基化分析(qAMP)检测组织DNA和ccfDNA中RERG mRNA甲基化率。绘制受试者工作特征(ROC)曲线评价ccfDNA中RERG mRNA甲基化率对鼻咽癌的诊断价值。结果基于TCGA数据库中来自于头颈部鳞状细胞癌(HNSC)组织标本的RNA-Seq转录本数据相应患者临床资料,HNSC组织标本中RERG mRNA表达明显低于正常组织,RERG mRNA甲基化β值高于正常组织(P<0.001)。在临床试验中,鼻咽癌组ccfDNA中RERG mRNA甲基化率高于对照组[0.127(0.095,0.175)vs.0.023(0.014,0.036),P<0.001]。经ROC曲线分析,检测ccfDNA中RERG mRNA甲基化率诊断鼻咽癌的曲线下面积(AUC)为0.990(95%CI:0.980~0.999)。ccfDNA和肿瘤组织中RERG mRNA甲基化率呈正相关(r=0.407,P<0.001)。鼻咽癌患者ccfDNA中RERG mRNA甲基化率在不同肿瘤T分期、N分期、TNM分期及EB病毒(EBV)DNA状态人群之间差异有统计学意义(P<0.05)。结论ccfDNA中RERG mRNA甲基化状态可能为鼻咽癌筛查中潜在的血液学生物标志物。Objective To investigate the clinical significance of Ras-related and estrogen-regulated growth inhibitor(RERG)methylation status detection in circulating cell-free DNA(ccfDNA)for the diagnosis of nasopharyngeal carcinoma.Methods A total of 100 patients diagnosed as the primary nasopharyngeal carcinoma in the hospital from January 2020 to June 2021 were collected as nasopharyngeal carcinoma group,while 97 noncancer volunteers were also recruited to this study as the control group.The DNA and ccfDNA in biopsy tissues were both extracted,and the methylation rate of RERG mRNA was detected by the quantitative analysis of DNA methylation using real-time PCR(qAMP).Receiver operating characteristic(ROC)curve was used to evaluate the predictive value of RERG methylation status in ccfDNA for nasopharyngeal carcinoma.Results Based on the clinical information of patients corresponding to the RNA-Seq transcript data from the head and neck squamous-cell carcinoma(HNSC)tissues samples in the TCGA database,the expression of RERG mRNA in HNSC tissues was significantly lower than that in normal tissues and the beta value of RERG mRNA methylation was also higher than that in normal tissues(P<0.001).In clinical trials,the methylation rate of ccfDNA RERG mRNA in nasopharyngeal carcinoma group was higher than that in control group[0.127(0.095,0.175)vs.0.023(0.014,0.036),P<0.001].ROC curve results showed that the area under curve(AUC)of methylation rate of RERG mRNA in ccfDNA for the diagnosis of nasopharyngeal carcinoma was 0.990(95%CI:0.980-0.999).The methylation rate of RERG mRNA in ccfDNA and tumor tissues was positively correlated(r=0.407,P<0.001).Besides,the differences of methylation rate of RERG mRNA in ccfDNA in patients with different status of T stage,N stage,TNM stage and EB virus(EBV)DNA were statistically significant(P<0.05).Conclusion The methylation status of RERG mRNA in ccfDNA may be a promising blood biomarker for nasopharyngeal carcinoma screening.

关 键 词：循环细胞游离DNA RAS样雌激素调节生长抑制因子 甲基化 鼻咽癌 实时荧光定量PCR法DNA甲基化分析 

分 类 号：R446.8[医药卫生—诊断学]