生物信息学分析miR-16-5p在乳腺癌中的作用及临床验证

Bioinformatics analysis and clinical validation of miR-16-5p in breast cancer

目的 生物信息学分析miR-16-5p在乳腺癌(BC)中的作用并进行临床验证。方法 BC的队列数据集来自高通量基因表达数据库(GEO)和癌症基因组图谱(TCGA),并通过生物信息学工具进行分析。在临床验证中,前瞻性纳入2018年1月至2022年2月接受手术的BC患者(BC组),获取BC组织和对应的癌旁正常组织。另纳入同期150例体检健康女性作为对照组,获取其血清样本,采用实时荧光定量PCR(qPCR)检测组织及血清miR-16-5p表达。所有BC患者至少随访3年。结果 共鉴定出195个差异表达基因(DEG),根据logFC≥1.5或logFC≤-1.5阈值,miR-16-5p表达显著下调,其受试者工作特征(ROC)曲线和Kaplan-Meier曲线分析显示,miR-16-5p区分BC组织及正常乳腺组织的曲线下面积(AUC)为0.73(95%CI:0.68~0.79),且miR-16-5p低表达的患者预后较差(P=0.013),因此miR-16-5p可作为候选miRNA。在临床验证集中,BC组织miR-16-5p相对表达量明显低于癌旁正常组织[0.46(0.29,0.66)vs. 0.82(0.56,1.08),P<0.001]。且BC患者血清miR-16-5p相对表达量也显著低于对照组[0.34(0.15,0.50)vs. 0.99(0.70,1.20),P<0.001]。ROC曲线显示,当血清miR-16-5p相对表达量≤0.520时可良好地诊断BC患者,AUC值为0.897(95%CI:0.863~0.932)。BC患者血清样本和BC组织样本中miR-16-5p相对表达量呈正相关(rs=0.373,P<0.001)。血清miR-16-5p表达与T分期、N分期、TNM分期有关(P<0.05)。血清miR-16-5p低表达组是影响BC患者患者3年内发生局部复发/远处转移及死亡的独立预测生物标志物(P<0.05)。结论 血清miR-16-5p表达对BC有诊断效能,低血清miR-16-5p表达预示着BC患者更差的疾病进展情况和3年内预后不良。

Objective To analyze the role of miR-16-5p in breast cancer(BC)by bioinformatics and to verify it.Methods The BC cohort data set was collected from the High throughput gene Expression Database(GEO)and the Cancer Genome Atlas(TCGA)and analyzed by bioinformatics tools.In clinical validation,BC patients(BC group)who underwent surgery between January 2018 and February 2022 were prospectively included to obtain BC tissue and corresponding paracancer normal tissue.Another 150 healthy women were included as control group.The serum samples were obtained.The expression of miR-16-5p in tissues and serum was detected by real-time quantitative PCR(qPCR).All BC patients were followed for at least 3 years.Results A total of 195 differense-expressed genes(DEGs)were identified.miR-16-5p expression was significantly down-regulated according to the threshold of logFC≥1.5 or logFC≤-1.5.Receiver operating characteristic(ROC)curve and Kaplan-Meier curve analysis showed that,The area under curve(AUC)of miR-16-5p to distinguish BC tissue from normal breast tissue was 0.73(95%CI:0.68-0.79),and patients with low expression of miR-16-5p had poor prognosis(P=0.013).Therefore,miR-16-5p could be used as a candidate miRNA.In the clinical verification set,the relative expression level of miR-16-5p in BC tissues was significantly lower than that in adjacent normal tissues[0.46(0.29,0.66)vs.0.82(0.56,1.08),P<0.001].The relative expression level of miR-16-5p in serum of BC patients was also significantly lower than that of control group[0.34(0.15,0.50)vs.0.99(0.70,1.20),P<0.001].ROC curve showed that when the relative expression level of serum miR-16-5p was less than 0.520,BC patients could be well diagnosed,and the AUC value was 0.897(95%CI:0.863-0.932).The relative expression level of miR-16-5p was positively correlated between the serum samples of BC patients and the tissue samples of BC patients(r s=0.373,P<0.001).The expression of serum miR-16-5p was correlated with the T stage,N stage and TNM stage(P<0.05).The low expression of serum miR-16-5p group was an independent predictor of local recurrence/distant metastasis and death in BC patients within 3 years(P<0.05).Conclusion Serum miR-16-5p expression is effective in the diagnosis of BC,and low serum miR-16-5p expression predicts worse disease progression and poor prognosis in BC patients within 3 years.