Atrophic gastritis and gastric cancer tissue miRNome analysis reveals hsa-miR-129-1 and hsa-miR-196a as potential early diagnostic biomarkers

摘要

BACKGROUND Gastric cancer(GC)is one of the most frequently diagnosed tumor globally.In most cases,GC develops in a stepwise manner from chronic gastritis or atrophic gastritis(AG)to cancer.One of the major issues in clinical settings of GC is diagnosis at advanced disease stages resulting in poor prognosis.Micro RNAs(mi RNAs)are small noncoding molecules that play an essential role in a variety of fundamental biological processes.However,clinical potential of mi RNA profiling in the gastric cancerogenesis,especially in premalignant GC cases,remains unclear.AIM To evaluate the AG and GC tissue mi RNomes and identify specific mi RNAs’potential for clinical applications(e.g.,non-invasive diagnostics).METHODS Study included a total of 125 subjects:Controls(CON),AG,and GC patients.All study subjects were recruited at the Departments of Surgery or Gastroenterology,Hospital of Lithuanian University of Health Sciences and divided into the profiling(n=60)and validation(n=65)cohorts.Total RNA isolated from tissue samples was used for preparation of small RNA sequencing libraries and profiled using next-generation sequencing(NGS).Based on NGS data,deregulated mi RNAs hsa-mi R-129-1-3 p and hsa-mi R-196 a-5 p were analyzed in plasma samples of independent cohort consisting of CON,AG,and GC patients.Expression level of hsa-mi R-129-1-3 p and hsa-mi R-196 a-5 p was determined using the quantitative real-time polymerase chain reaction and 2-ΔΔCt method.RESULTS Results of tissue analysis revealed 20 differentially expressed mi RNAs in AG group compared to CON group,129 deregulated mi RNAs in GC compared to CON,and 99 altered mi RNAs comparing GC and AG groups.Only 2 mi RNAs(hsa-mi R-129-1-3 p and hsa-mi R-196 a-5 p)were identified to be step-wise deregulated in healthy-premalignant-malignant sequence.Area under the curve(AUC)-receiver operating characteristic analysis revealed that expression level of hsa-mi R-196 a-5 p is significant for discrimination of CON vs AG,CON vs GC and AG vs GC and resulted in AUCs:88.0%,93.1%and 66.3%,respectively.Comparing results in tissue and plasma samples,hsa-mi R-129-1-3 p was significantly down-regulated in GC compared to AG(P=0.0021 and P=0.024,tissue and plasma,respectively).Moreover,analysis revealed that hsa-mi R-215-3 p/5 p and hsa-mi R-934 were significantly deregulated in GC based on Helicobacter pylori(H.pylori)infection status[log2 fold change(FC)=-4.52,P-adjusted=0.02;log2 FC=-4.00,P-adjusted=0.02;log2 FC=6.09,P-adjusted=0.02,respectively].CONCLUSION Comprehensive mi RNome study provides evidence for gradual deregulation of hsa-mi R-196 a-5 p and hsa-mi R-129-1-3 p in gastric carcinogenesis and found hsami R-215-3 p/5 p and hsa-mi R-934 to be significantly deregulated in H.pylori carrying GC patients.