Combined Identification of Three miRNAs in Serum As Effective Diagnostic Biomarkers for HNSCC

Abstract

Background: Head and neck squamous cell carcinoma (HNSCC) is a disastrous disease with substantial morbidity and mortality. This study aims to explore the effective diagnostic and prognostic biomarker for HNSCC. Methods: MiRNA expression data and corresponding clinical information of HNSCC from The Cancer Genome Atlas (TCGA) database were analyzed comprehensively to identify the differentially expressed miRNAs. The predictive ability of different classification was analyzed of the three-miRNA combinations. Then, diagnostic and prognostic value were evaluated and verified in clinical patients. Results: A total of 486 HNSCC samples and 44 normal samples were obtained from the TCGA database. After analysis, 128 differentially expressed miRNAs in HNSCC tissues were identified, and 10 miRNAs were finally selected for further study. Classification analysis developed a threemiRNA signature of hsa-mir-383, hsa-mir-615 and hsa-mir-877 with an average AUC (Area Under the Curve) of 99.4% in differentiating tumor tissues from normal tissues. The combination of the three miRNAs in serum yielded an AUC value of 0.986 (95%CI, 0.974-0.998; P＜0.0001) with 97.5% sensitivity and 94.6% specificity, which was superior to that of a single or a two-miRNA signature. One-way ANOVA test results suggested the expression levels of hsa-mir-615 and hsa-mir-877 were significantly changed with different tumor grades, with the p-values of 0.001 and 0.056, respectively, while hsa-mir-383 showed no significant correlation with tumor grade (P=0.18). And no significant correlations were found between hsa-mir-383, hsa-mir-615 or hsa-mir-877 and tumor stage with the p-values of 0.14, 0.95 and 0.81, respectively. Survival analysis indicated different expression levels of hsa-mir-383 led to significantly different survival rates (P<0.022), while no significant survival differences were observed between the high and low expression set of hsamir-615 (P<0.401) or hsa-mir-877 (P<0.403). This result was verified in 81 validation patients. The low expression level of hsa-mir-383 was significantly correlated with poor prognosis (P=0.0404), while the expression level of hsa-mir-615 or hsa-mir-877 showed no significant relationship with the prognosis (hsa-mir-615, P=0.6247; hsa-mir-877, P=0.5199). Conclusion: A three-miRNA signature of hsa-mir-383, hsa-mir-615, and hsamir-877 may serve as an excellent diagnostic biomarker for HNSCC, and therein hsa-mir-383 shows potential prognostic significance for HNSCC patients. Funding Statement: Shandong Provincial Health Commission Foundation (No. 2016WS0410), China postdoctoral Scinence Foundation Grant (No. 2019M652380), Fundamental Research Funds of Shandong University (No.2018CJ047). This work is also supported by Qilu Young Scholar Program of Shandong University. Declaration of Interests: The authors declare that they do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted. Ethics Approval Statement: All patients and normal controls gave informed consent. This study was approved by the Medical Institutional Ethical Committee of the Provincial Hospital Affiliated to Shandong University.