The Target Reseach Of MiR-103a-3p And Dicer1 In Pancreatic Carcinoma Panc-1 Cell

Objective: Micro RNAs(mi RNAs) have bearing on the occurrence and development of tumors, which lead to degrade or inhibit translation of the target genes by binding to 3’UTR of m RNA targets. In the end, mi RNAs promote or suppress the occurrence and development of tumors. In this study we search for potential target genes of mi R-103a-3p and validate the targeting relationship of mi R-103a-3p and potential target genes in pancreatic carcinoma Panc-1 cell line. On this basis, we probe into the way that mi R-103a-3p act on Dicer1 further. so as to provide the foundation for the vivo tests, and theoretical basis of the effective target for therapy in pancreatic carcinoma.Methods: 1. To screen the potential target genes of mi R-103a-3p by several prediction tools. After a comprehensive analysis, we chose Dicer1 from predicted target genes for further study; 2. The relationship of mi R-103a-3p and Dicer1 was revealed by Dual-Luciferase Reporter Assay System; 3. The mi R-103a-3p inhibitor was transfected transiently into pancreatic carcinoma Panc-1 cell lines. Then, the transfection efficiency would be tested; 4. The expression of mi R-103a-3p and Dicer1 was detected by QPCR and western blot, identifying the Dicer1 as target of mi R-103a-3p by function.Results: 1. The gene of Dicer1 which predicted by eight databases had the high predictive values and popular reseach in pancreatic carcinoma. So we choose the Dicer1 as mi R-103a-3p targets; 2. The result of the detected gene sequence, the GV306-Dicer1-3’UTR vector was successfully constructed. In Panc-1 cell lines, Luciferase reporter system showed that the luciferase activity was increased significantly respectively in Control group compared to Blank or NC group after transfection of Dicer1-3’UTR plasmid and mi R-103a-3p inhibitor(P<0.05); 3. The mi R-103a-3p expression was decreased by transfected the mi R-103a-3p inhibitor into Panc-1(P<0.05); 4. The concentration of mi R-103a-3p was significantly correlated with Dicer1 protein while it was negatively correlated with the Dicer1 m RNA.Conclusions: 1. The Dicer1 as mi R-103a-3p targets, there is binding site of mi R-103a-3p in the 3’-UTR of Dicer1; 2. The pancreatic carcinoma Panc-1 cell lines of the low expression of mi R-103a-3p had been successfully constructed by the mi R-103a-3p inhibitor transfected transiently; 3. In Panc-1 cell lines, mi R-103a-3p regulate the expression of Dicer1 by post-transcriptional level.