Expression of miRNA of serum glycosylated exosomes in pancreatic ductal adenocarcinoma and its clinical significance

MIAO Yongli; LIANG Juan; ZHANG Qian

doi:10.3969/j.issn.1009-0460.2025.06.007

Abstract：Objective To investigate the expression of miRNA in serum glycosylated exosomes of pancreatic ductal adenocarcinoma(PDAC)and its relationship with clinicopathological characteristics and prognosis.Methods From March 2020 to July 2023,a total of 118 patients with PDAC(PDAC group),98 patients with benign pancreatic diseases(BDG group),and 85 healthy volunteers(HC group)were included and assigned to the discovery,screening,and validation cohors.Fucosylated exosomes were isolated from serum by the lentil lectin magnetic bead method.miRNA sequencing was performed on the serum samples in the discovery set(n=24).Twelve differentially expressed mirnas(DEMirs)were selected for screening in the screening set(n=60)by real-time fluorescence quantitative PCR(qPCR),and two candidate mirnas were selected for further verification in the validation set(n=216).Analyze the relationship between miRNA expression and the clinicopathological characteristics and overall survival period of patients.Results Serum glycosylated exosomes were successfully isolated through the fucose capture strategy based on lentil lectin magnetic beads.miRNA sequencing was performed on the samples of the discovery set,and a total of 36 overlapping DEMirs were identified in the three groups.The screening set confirmed that only the levels of miR-199a-3p and miR-421 in exosomes differed among the three groups(P<0.001).In the validation set,these two candidate mirnas were the highest in the PDAC group[miR-199a-3p:2.76(1.74,4.36);miR-421:2.35(1.38,4.49)],followed by the BDG group[miR-199a-3p:1.94(0.99,2.52);miR-421:1.44(0.78,1.77)],and the HC group was the lowest[miR-199a-3p:0.75(0.58,1.09);miR-421:0.97(0.46,1.49)].miR-199a-3p+miR-421+CA19-9 can significantly improve the accuracy of differential diagnosis or prediction of PDAC vs.BDG and PDAC vs.BDG+HC,and the areas under the receiver operating characteristic curves are 0.927 and 0.958,respectively.miR-199a-3p was associated with smoking history(P=0.048),TNM stage(P=0.005),and lymph node metastasis(P=0.002).miR-421 was associated with body mass index(P=0.020),lymph node metastasis(P=0.030),distant metastasis(P=0.011),and degree of differentiation(P=0.004).The median follow-up time was 12.0 months,and the 1-year mortality rate was 53.49%.The median overall survival time in the miR-199a-3p high-expression group(9.21 months vs.23.52 months)or the miR-421 high-expression group(8.05 months vs.24.02 months)was shorter than that in the low-expression group(P<0.001).Analysis by the Cox proportional hazards regression model showed that serum exosome miR-199a-3p and miR-421,as well as history of alcohol consumption,history of diabetes,TNM stage,tumor diameter,and distant metastasis were independent factors affecting overall survival(P<0.05).Conclusion Serum glycosylated exosomes miR-199a-3p and miR-421 are promising non-invasive biomarkers for PDAC.