DC’s Proliferation And Differentiation And The Influence Of Expression For MiRNA-146a、c-jun And NF-κB In The Microenvironment Of Pancreatic Cancer

Objective: Pancreatic carcinoma is a malignancy with extreme poor prognosis,and the incidence and mortality are increasing every year. At present, treatmentcombined surgery, chemotherapy and radiotherapy is the main, but the effect is poor.DC are the most powerful antigen presenting cells, playing an important role in antigensubmission, T cell activation and immune tolerance. According to research findings,DC’s function and quantity of Pancreatic cancer patient’s peripheral blood are obviouslylimited, which are closely related to the cancer subtypes and degree of malignancy.NF-kB is a transcription factor widely existed in eukaryotic cell, playing an importantrole in immune, tumor, inflammation and many other diseases. In the transcriptionregulation, it can be combined with a sequence of kB of a variety of protein genepromoter and enhancer, inducing these protein expression enhancement, and producinga series of biological effect. As an important nucleoprotein, NF-kB’s abnormal activityincreases the happening and developing of the neoplastic disease. As an earlyprotocarcinogenic gene, c-jun’s protein product is part of the AP-1(activating protein-1)transcription factor, playing a role in gene transcription of a variety of cell factors andgrowth factors, regulating the cell growth, proliferation and apoptosis. DC vaccine isone of the hot spot for studying the tumor. Many studies show that miRNA-146a plays arole in many signal transduction, participating in the origin and development of thetumor. Also studies show that c-jun and NF-kB play an important role in DC’s processof maturation. To study the DC’s proliferation and differentiation and the influence ofexpression for miRNA-146a, c-jun and NF-kB in the microenvironment of Bxpc-3Pancreatic cancer, and explore the microenvironment of Bxpc-3pancreatic cancer toDC’s mechanism of action, providing a theoretical basis for DC vaccine treatingPancreatic cancer. Methods: We cultured the Bxpc-3cells and got the supernatants fluid to simulatethe microenvironment of pancreatic cancer tumor. Peripheral blood of healthyvolunteers collected sterile, with heparin, mononuclear cells were isolated withlymphocyte separation medium. To isolate the peripheral blood mononuclear cells andculturing, adding CSF, IL-4, TNF-α to get the DC. We divided the DC into four groupsincluded imDC, imDC+supernatants fluid of Bxpc-3, mDC and mDC+supernatantsfluid of Bxpc-3. Phenotype (CD1a,CD14,CD80,CD86) were identified by flowcytometry. The expression of miRNA-146a was examined by q-PCR. The expression ofc-jun and NF-kB were determined by western blot.Results: The expression of CD1a in supernatants fluid of Bxpc-3group, as humanperipheral blood mononuclear cells sign, was significantly fall. As mature DC’s sign,CD80and CD86in supernatants fluid of Bxpc-3group were significantly fall. Asimmature DC’s sign, CD14was conspicuously high. We examined the expression ofmiRNA-146a by q-PCR, the level of miRNA-146a in supernatants fluid of Bxpc-3groups was overexpressed. We determined the expression of c-jun and NF-kB bywestern blot, the expression level of c-jun and NF-kB in supernatants fluid of Bxpc-3groups was all reduced(P＜0.05).Conclusion: The supernatants fluid of Bxpc-3in DC can increase the expressionlevel, decline the c-jun and NF-kB protein expression, then inhibiting the maturationprocess of DC, and reducing DC’s damagment for tumor cell.