miR-101基因簇与食管癌发病风险的病例对照研究

A case-control study on expression of miR-101 cluster and risk of esophageal carcinomas

目的:探讨miR-101基因簇与食管癌发病风险的关系。方法:收集3例食管癌进展期患者的新鲜组织标本进行组织芯片高通量microRNA检测,采用肿瘤基因组图谱(TCGA)数据库食管癌RNA测序数据验证的方式对miR-101基因簇表达水平进行分析;另收集33例食管癌患者癌组织和癌旁组织标本,分别提取样本总RNA,采用实时荧光定量PCR法(qPCR)检测miR-101基因簇相关基因的表达,logistic回归分析miRNA异常表达对食管癌发病风险的影响,受试者特征曲线(ROC)评价miR-101基因簇对食管癌的诊断效力,Fisher检验分析基因簇各miRNA的表达和食管癌患者临床病理指标之间的相关性。结果:miRNA芯片高通量检测及TCGA数据库生物信息学分析结果显示,与食管正常组织相比,miR-101-3p、miR-125a-5p和miR-145-5p在食管癌组织中表达下调(P<0.05)。q PCR结果显示miR-101基因簇在33例食管癌组织中均呈低表达,多因素logistic回归分析显示,miR-101基因簇miR-101-3p、miR-127-5p与miR-145-5p表达水平与食管癌的发病风险呈负相关[OR(95%CI)值分别为:0.717(0.563,0.912)、0.717(0.534,0.962)和0.597(0.426,0.838),均为P<0.05];ROC分析显示miR-101基因簇曲线下面积(AUC)分别为0.753、0.792、0.763和0.800(P<0.05)。此外,miR-101-3p、miR-145-5p的表达水平与食管癌患者肿瘤大小相关(P<0.05),miR-125a-5p的表达水平与患者淋巴结转移相关(P<0.05)。结论:miR-101基因簇在食管癌患者癌组织中呈低表达,可作为食管癌诊断的潜在生物标志物,其在食管癌进程中的功能和调控机制有待进一步研究。

OBJECTIVE:To investigate relationships between expression of mi R-101 gene cluster and risk of esophageal cancer.METHODS:Expression levels of the mi R-101 gene cluster was analyzed using high-throughput detection mi RNA gene chips and was verified using esophageal cancer RNA sequencing data in the cancer genome atlas(TCGA)database.In addition,total RNA was extracted from 33 patients’esophageal cancer tissues and adjacent tissues.These RNA samples were used to detect expression of the mi R-101 gene cluster using real-time fluorescent quantitative PCR(q PCR).Relationships between abnormal expression of mi RNA and risk of esophageal cancer were analyzed by logistic regression.The diagnostic efficacy of mi R-101 gene cluster on esophageal cancer was evaluated using receiver operator characteristics curve(ROC).The Fisher test was used to analyze correlations between expression of mi RNA in gene cluster and clinicopathological factors in the patients.RESULTS:High-throughput detection of mi RNA chip and bioinformatics analyses of TCGA database showed that the expressions of mir-101-3p,mir-125a-5p and mir-145-5p were down regulated in esophageal carcinomas compared with normal esophageal tissues(P<0.05).q PCR results showed that expressions of the mi R-101 gene cluster were low in esophageal cancers.Multivariate logistic regression analyses showed that expression levels of mir-101-3p,mir-127-5p and mir-145-5p were negatively correlated with the risk of esophageal cancer[OR values were 0.717(0.563,0.912),0.717(0.534,0.962)and 0.597(0.426,0.838),P<0.05].ROC analyses showed that AUCs of the mi R-101 gene cluster was 0.753,0.792,0.763 and 0.800,respectively(P<0.05).In addition,expression levels of mir-101-3p and mir-145-5p were correlated with tumor size(P<0.05),and that of mir-125a-5p were correlated with lymph node metastasis(P<0.05).CONCLUSION:Expression of the mi R-101 gene cluster was low in esophageal cancer tissues compared to adjacent normal tissues.The expressions can be used as dpotential biomarker for the diagnosis of esophageal cancer but their functions and regulatory mechanisms in the cancer process need to be further studied.