PAMD Alters The Expression Profiles Of Micrornas In BxPC-3 Human Pancreatic Carcinoma Cells

Purpose:The present paper attempted to explore that PAMD alters the expression profiles of miRNAs in BxPC-3 human pancreatic carcinoma cells.Methods:BxPC-3 human pancreatic carcinoma cells were cultured with PAMD final concentration of 80μg/ml(P80 group).The blank control group was cultured in normal culture medium.The following experiments were carried out:1.To verify the anti-cancer effects of PAMD via a miRNAs-dependent mechanism,comprehensive miRNAs expression profiling of PAMD-treated BxPC-3 human pancreatic cancer cells Homogenization and miRNAs labeling.2.To ensure that reagents the hybridization mix containing PAMD treatment group and control group mixing in a PCR tube.3.Microarray assay scanning and analysis based on the Axon GenePix 4000B microarray scanner,to read the raw intensity of the image by GenePix pro V6.0.4.MicroRNAs target prediction and Gene Ontology and KEGG pathway analysis were performed to investigate possible pathways involved.5.Using real-time quantitative PCR method to verify the results of microarray.Results:1.The results showed that 301 miRNAs were significantly differentially expressed(FC>2 or FC<0.5,P<0.05)in PAMD-treated BxPC-3 human pancreatic cancer cells.2.The prediction of miRNAs targets was performed using the online software TargetScan,PicTar and miRanda.The intersection of the results from these three types of software was taken as the final target genes of significantly differentially expressed miRNAs.Inferring PAMD alters the expression profiles of miRNAs in BxPC-3 human pancreatic carcinoma cells.3.GO and KEGG pathway analyses were performed on the target genes of the significantly differentially expressed miRNAs.The results presented revealed that the influence of BxPC-3 pancreatic cancer cells by PAMD may be related to neuroactive ligand-receptor interactions,pathways involved in cancer,MAPK signalling pathways,focal adhesion,calcium signalling pathways and other factors.Conclusion:Our data indicates that PAMD inhibits BxPC-3 cells probably through regulating the expression of miRNAs.Interruption of miRNAs profiling may provide new therapeutic methods for the clinical treatment of pancreatic cancer.