| Background:Pancreatic cancer is a kind of high malignant degree of tumor, and the incidence is increasing, the prognosis is poor, the 5-year survival rate is low, the main reason is the existing diagnosis of pancreatic cancer markers of specificity and sensitivity is not high, the early diagnosis rate is low, cannot be timely effective treatment early on. Reportedly the serum exosomal miRNA (exosomal miRNA) expression level of change and the correlation with clinical pathologic factors such as pancreatic cancer stage, differentiation, is a kind of good serological tumor markers, but in pancreatic cancer on serum exosomal microRNAs reported fewer still. This study aims to through the in vitro experiment, animal experiment and clinical case analysis to find significantly associated with pancreatic cancer in serum exosomal miRNA biomarkers.Objectives:find associated with pancreatic cancer serum exosomal miRNA biomarkers.Methods:(1)experiment in vitro:in vitro pancreatic cancer cell lines (PANC-1),By filtering centrifuge technology to extract the exosome, exosome PCR Array to detect PANC-1 and 84 kinds of pancreatic cancer related miRNA expression level and variance analysis;(2) Animal experiment:Pancreatic cancer nude mouse model is established using the PANC-1, respectively for the experimental group (group A) and control group (group B) serum samples and samples of tumors had tumor model, extract serum exosome, through RT-PCR detection in group A and group B mice exosomal miR 202-3p, miR-101-3p of 9 kinds of high expression levels of serum exosomal microRNAs. Test group 20 serum samples exosomal miR-202-3p expression level, and analyzes its relationship with tumor weight.detection of group A and group B in the mice miR-202-3p, miR-101-3p and other 9 kinds of high expression levels of serum exosome micrornas.In the test group B 20 serum samples exosome miR-202-3p expression level, and analysis Its relationship with tumor weight.(3) the clinical experiment:Clinical experiment:choose two with pancreatic cancer significantly close exosomal serum miR-202-3p and miR-101-3p as test object. Collect patients with pancreatic cancer and pancreatic cancer patients serum, by RT-PCR technique to detect serum exosomal miR-202-3p and expression levels of miR-101-3p, with clinical pathologic factors such as pancreatic cancer staging and differentiation of correlation analysis to evaluate the value of the clinical diagnosis of pancreatic cancer.Results:(1) the exosome PCR Array to detect PANC-1 in 84 pancreatic cancer related micrornas, screening out the miR-202-3p, miR-101-3p of 9 kinds of relatively high expression of exosomal miRAN; (2)Tumor animal model group nine exosome miRAN expression levels were higher than in normal control group, miR-202-3p and miR-101-3p the expression level of 2.33 and 2.03 times higher respectively than control group; And with the increase of the tumor, miR-202-3 p expression level increased; (3) pancreatic cancer group exosomal in serum miR-202-3p and miR-101-3p expression level was significantly higher than that of non pancreatic cancer group, the serum exosomal miR-202-3p expression level and no correlation between pancreatic cancer staging and distant metastasis.Conclusions:(1) Experiments show that serum tests outside secrete tiny RNA method can be used in clinical diagnosis of pancreatic cancer; (2) can be detected in pancreatic cancer animal model was relatively high expression of exosomal microRNAs, miR-202-3p and miR-101-3p abnormally high expression, and with the tumor progression, miR-202-3p expression level increased, is helpful to guide the diagnosis of pancreatic cancer associated with progression of clinical research; (3) in pancreatic cancer has a high expression level of exosomal miR-202-3p and miR-101-3p can be used for clinical diagnosis and differential diagnosis of pancreatic cancer markers, and exosomal miR-202-3p abnormal expression for clinical staging of pancreatic cancer, the grasp of the surgical indications and prognosis judgement has very important guidance value. |
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