Abstract
OBJECTIVES: Esophageal cancer (EC) is a highly aggressive malignancy with a low survival rate. Current detection methods like endoscopy are invasive and expensive, resulting in low compliance rates. This study aimed to develop and validate a novel circulating microRNA (miRNA)-based panel for early EC detection and compare its effectiveness in serum versus plasma samples. METHODS: The study developed a multiplexed miRNA assay to detect eight EC-related miRNAs in both plasma (n=45) and serum (n=148) samples. Expression levels of these miRNAs were compared between EC patients and control subjects. The serum-based panel was further validated in an independent cohort and compared with carcinoembryonic antigen (CEA), a conventional tumor marker. RESULTS: The multiplexed miRNA panel demonstrated excellent PCR amplification efficiency with no cross-interference between miRNAs. Six of the eight miRNAs showed significantly higher expression in EC patients compared to controls in serum samples (area under the curve (AUC) >0.900), while plasma samples showed poor discrimination (AUC<0.700). The optimized 6-miRNA panel in serum achieved perfect discrimination (AUC=1.0) between EC and controls in both training and validation cohorts. This panel also effectively distinguished EC from gastric and colorectal cancers and significantly outperformed CEA (AUC=0.619) in diagnostic accuracy. CONCLUSION: Serum is more suitable than plasma for EC detection using miRNA biomarkers. The optimized 6-miRNA panel demonstrated exceptional accuracy for EC detection, offering a promising non-invasive approach for early diagnosis. This method requires smaller sample volumes than traditional techniques, potentially improving clinical applicability and patient compliance.