The Function Of Exosomal MicroRNA In Diagnosis And Prognosis Of Pancreatic Cancer Using A Tethered Cationic Lipoplex Nanoparticle Biochip

Background:Due to the lack of reliable and sensitive biomarkers for early diagnosis,most patients with pancreatic cancer(PC)are in an unresectable status and lose the opportunity for surgery once diagnosed,which is considered to be one of the most important reasons for the high mortality of patients with PC.Recently,various studies showed that exosomal micro RNA(exmiR)palyed an important role in the diagnosis and prognosis of malignant tumors.Relevant literatures and our previous studies have shown that miR-21,miR-10 b,miR-212-3p,and miR-18a-5p were highly expressed in tumors and have certain value for tumor diagnosis and prognosis.Objective:(1)To analyze and compare the expression levels of exmiR-21,exmiR-10 b,exmiR-212-3p and exmiR-18a-5p in different stages of PC group,healthy controls group and benign pancreatic disease group;(2)To analyze and compare the value of exmiR in the diagnosis of PC when used alone and in combination,and select exmiR that can be used for the diagnosis of PC;(3)To analyze and compare the relationship between the expression level of exmiR and the prognosis of PC,and select the exmiR that can predict the prognosis of PC.Methods:(1)We evaluated the plasma expression of exmiR-21,exmiR-10 b,exmiR-212-3p,and exmiR-18a-5p in 36 PC patients,36 healthy individuals,and 8 patients with benign pancreatic disease by using the tethered cationic lipoplex nanoparticle(TCLN)biochip.To compare the expression levels in different groups.(2)Using receiver operating characteristic(ROC)analysis,the area under the ROC(AUC)curve was calculated.To compare the value of each exmiR in the diagnosis of PC when used alone or in combination with others.(3)Univariate Cox regression analysis was used to screen the potential clinical factors and exmiRs that affect the prognosis of PC.Multivariate Cox regression analysis was used to evaluate independent risk factors affecting PC prognosis.Results:(1)Compared with healthy individuals,the expression level of exmiR-21 was significantly increased in patients with PC(TNM ?-?)(p <0.0001).The levels of exmiR-21 in patients with TNM stage ? and ? and those in stage ? and ? were significantly higher than those of in healthy controls(p = 0.0115;p = 0.0003).The expression level of exmiR-10 b in PC group was significantly higher than that in healthy group(p = 0.0025).The expression level of plasma exmiR-18a-5p in PC group was significantly higher than that in healthy group(p = 0.0002).There was no statistically significant difference in plasma exmiR-212-3p expression in healthy individuals,benign pancreatic disease group,and PC group.(2)(2)ROC analyses showed that expression level of exmiR-21 had a AUC of 0.82(p <0.0001)in PC group and healthy controls,and its sensitivity was 69.4% and specificity was 100%.In PC group and the pancreatic benign disease group,the AUC was 0.75(p = 0.0370),the sensitivity was 80.5%,and the specificity was 75%.In healthy controls and the benign pancreatic disease group,the AUC was 0.54(p = 0.7150).The AUC of exmiR-18a-5p and exmiR-10 b in PC group and healthy controls were 0.79(p <0.0001)and 0.75(p = 0.0003),and their corresponding sensitivity and specificity were 63.9%,100% and 55.6%,86.1%.ExmiR-212-3p was incapable of identifying the PC patients from the health(AUC =0.56,p = 0.3499),but it could distinguish the PC group and the pancreatic benign disease group(AUC = 0.73,p = 0.0446;sensitivity 52.8%,Specificity 87.5%),benign pancreatic disease group and healthy control group(AUC = 0.73,p = 0.0446;sensitivity:75%,specificity: 77.8%).The participants were further divided into two groups,PC group and non-PC group.The results show that exmiR-21,exmiR-18a-5p,and exmiR-10 b could distinguish patients in PC group and in NPC group(AUC = 0.81,p<0.0001,sensitivity 69.4%,specificity 95.5%;AUC = 0.81,p <0.0001,sensitivity55.6%,specificity 81.8%;AUC = 0.72,p = 0.0007,sensitivity 63.9%,specificity97.7%).ROC analysis of exmiR-21 combined with exmiR-18a-5p showed that AUC =0.84(p <0.0001),sensitivity was 75%,and specificity was 95.5%.ROC analysis was performed for ExmiR-21 combined with exmiR-10 b and exmiR-10 b combined with exmiR-18a-5p.The AUC were 0.84(p <0.0001)and 0.80(p <0.0001),respectively.Its sensitivity was 75% and 63.9%,and specificity was 93.2% and 97.7%,respectively.Analysis of three exmiRs combined with ROC showed that AUC = 0.84(p <0.0001),with a sensitivity of 75% and specificity of 95.5%.(3)Univariate Cox regression analysis showed that age(p=0.031),the TNM stage(p=0.037),surgery(p=0.031),and the expression of exmiR-212-3p(p=0.020)were potential prognostic factors of PC patients.Multivariate Cox regression analysis suggested that TNM stage in ? and ?(p=0.023)and low expression of exmiR-212-3p(p=0.011)were independent risk factors affecting the prognosis of patients with PC,and related to better prognosis.Conclusion:(1)The results of our study showed that exmiR-21,exmiR-10 b,and exmiR-18a-5p levels were increased in patients with PC.ROC analyses showed that exmiR-21 had the highest accuracy and sensitivity as a single diagnostic marker among the four candidate exmiRs,and was a potential biomarker for early diagnosis of pancreatic cancer.ExmiR-21 combined with exmiR-18a-5p could improve the diagnostic efficacy and the sensitivity of pancreatic cancer diagnosis.(2)TNM stage of PC and expression level of exmiR-212-3p were independent risk factors that affect the prognosis of patients withPC,and stage I and II,and low expression level of exmiR-212-3p was related to better prognosis of PC patients.