Potential miRNA biomarkers for the diagnosis and prognosis of esophageal cancer detected by a novel absolute quantitative RT-qPCR method

Zhiyuan Lin,Jiajia Wang,Zanxi Fang,Yongquan Chen,Jing Teng,Xiaosong Su,Zhongying Zhang,Huayue Lin,Qingchun Wei,Hongwei Li,Yanling Lin

doi:10.1038/s41598-020-77119-6

Abstract

miRNAs are expected to become potential biomarkers in the diagnosis and prognosis of Esophageal cancer (EC). Through a series of screening, miR-34a-5p, miR-148a-3p and miR-181a-5p were selected as EC-associated miRNAs. Based on AllGlo probe, a novel absolute quantitative RT-qPCR method with high sensitivity, specificity and accuracy was established for detecting miRNAs. Then the clinical significance of these 3 miRNAs was explored with 213 patients (166 cases with EC and 47 cases with benign diseases) and 170 normal controls. Compared with normal controls, the level of miR-34a-5p increased while miR-148a-3p and miR-181a-5p decreased in EC and benign patients (P < 0.001), and the level of miR-181a-5p in early EC patients was significantly lower (P < 0.001). According to logistic regression analysis, combined detection of miR-34a-5p, miR-148a-3p and Cyfra21-1 provided the highest diagnosis efficiency of 85.07% with sensitivity and specificity reaching 85.45% and 84.71%. Compared with preoperative samples, the level of miR-34a-5p decreased while miR-148a-3p and miR-181a-5p increased in postoperative samples (P < 0.001). Collectively, this first developed, novel absolute quantitative RT-qPCR method exhibits high application value in detecting miRNAs, miR-34a-5p, miR-148a-3p and miR-181a-5p may serve as potential biomarkers in the diagnosis and prognosis of EC, and miR-181a-5p probably could serve as a new biomarker for early EC.

Highlights

Esophageal cancer (EC), one of the most common malignant tumors in the clinic, ranks the sixth leading cause of cancer mortality in the world with progressive dysphagia as the main clinical manifestation[1]
Establishment and evaluation of the novel absolute quantitative reverse transcription (RT)‐qPCR method based on AllGlo probe for detecting miRNAs
Using validated specific primers and designed probe sequence, the absolute miRNA quantitative RT-qPCR detection method based on AllGlo probe was constructed

Summary

Introduction

Esophageal cancer (EC), one of the most common malignant tumors in the clinic, ranks the sixth leading cause of cancer mortality in the world with progressive dysphagia as the main clinical manifestation[1]. The invasiveness, high cost and the missed diagnosis of early patients make it unable to be used as a common screening method for physical examination. Studies have shown that the expressions of miRNAs are involved in the developments, invasion and metastasis of several kinds of cancers, including colorectal c ancer[10,11], breast c ancer[12,13], lung c ancer[14], which are hopefully to become potential biomarkers in plasma in the diagnosis and therapy. Considering the expression levels of miRNAs were different in the stages of development, invasion and metastasis of many other cancers, we decided to explore whether the abundance of m iRNAs15,16 was changed in the plasma of EC patients and whether the miRNAs could serve as biomarkers for EC. The clinical diagnosis significance of the screened miRNAs and their combined models in the diagnosis and prognosis of EC were explored

Methods

Results

Conclusion