Study On The Expression Level Of MiRNA Markers And Their Target Genes From Peripheral Blood Leukocyteof Patients With Esophageal Cancer

Objective:By screening and validating the peripheral blood microRNA markers of esophageal cancer patients,predicting their target genes,and detecting the expression of the corresponding mRNA and protein in esophageal cancer patients,to explore the possible relationship between their abnormal expression and esophageal cancer.Methods:Taking the patients with esophageal carcinoma as the resources,and taking the healthy group as the normal control group,using miRNA chip technology and real time PCR,to determine the peripheral blood leukocyte miRNA of patients with esophageal carcinoma,Online Software like TargetScan,miRDB,PicTar are used to predict the possible target gene of miRNA.Their function related to tumor,immune,cell cycle,cell proliferation and cell differentiation is retrieved from the Pubmed database,the expression level of target genes are detected by qPCR and Western Blot.Results:The results of microarray screening showed that 184 miRNAs were up-regulated and 143 miRNA were down-regulated in esophageal carcinomacompared with the normal control group.Compared with the normal control group,the qPCR results showed that the expression level of mir-181b-5p and mir-4695-3p in patients with esophageal carcinoma was down-regulated(p<0.05)and the expression level of miR-4725-5p was up-regulated(p<0.001).Through on-line prediction and offline verification of the targrt genes of above three kinds ofmiRNAs,we found that compared with normal control group(1.4±0.31),the mRNA expression level of YWHAH gene was up-regulated in esophageal carcinoma group(4.73±1.17),and there was statistical difference between them(P < 0.01);The mRNA expressionlevel of COL1A1 gene was up-regulated in esophageal carcinoma group(P <0.05),while its expression in normal group(1.82±0.39),esophageal carcinoma group(7.76±1.86);Compared with normal group(2.20±0.50),the mRNA expression level ofHINT1 gene was down-regulated in esophageal cancer group(0.93±0.17),there was statistical difference(P<0.05).And the result of WB shows that compared with normal group,the expression level of YWHAH and COL1A1 were up-regulated in esophageal cancer group.Conclusion:1.According to RT-qPCR result,MiR-181b-5p,miR-4695-3p and miR-4725-5p may be the potential molecular markers for early diagnosis of esophagealcancer.2.There might be a regulatory relationships between miR-181b-5p,miR-4695-3p and target genes YWHAH,COL1A1.