Study On MiR-15a Mechanism In Influenza Virus Inducing Apoptosis

Objection:the experiment was to study human lung adenocarcinoma epithelial cell model infected by influenza virus. Using Real-time PCR technology, over-expression and antisense technology, and bioinformatics analysis techniques identified correlation mechanism of objection miRNA and influenza virus infection. Anylysis the objection miRNA mechanism in influenza virus infection process.Methods:It is Real-time PCR to detect host cell infected by influenza virus miR-15a expression. Host cell miR-15a expression was influenced by antisense technology and over-expression. And then Cell morphology, measurement of living cells, Virus multiplication and apoptosis detected by flow cytometry discussed the contact of influenza virus pathogenesis and miR-15a. According to relevant research results that had been tested, the study Identified known miR-15a target genes expression and miR-15a new target genes.The study further illustrated influenza virus infection pathogenic mechanismResults:The experiment successfully constructed miR-15a eukaryotic expression vector. PCR and DNA sequence identification was consistent with purpose gene sequences. Real-time PCR tests showed that miR-15a expresseion was significantly higher than the negative control plasmid expression. Difference showed a Statistical significance (P<0.05).Real-time PCR results showed that miR-15a expression was significantly increased after A549 Cells transfecting influenza virus (P<0.05).After Excessive expressions and inhibition miR-15a, CCK-8 method test indicated that Along with miR-15a inhibitor dose reduction, the cell pathologic rates was increasing.the test demonstrate miR-15a could induce apoptosis.Flow cytometric-Annexin V/PI double dyeing method test showed that with the miR-15a inhibitor dose increasing cell lesions rate redeced. Test results showed that miR-15a had participated the progress of the influenza virus induced A549 cells apoptosis.mRNA and protein level expression results of miR-15a target gene showed BCL2,CCND1 and P85 target genes expression gradually dereased as time 0h,12h, 24h.No visible difference of action showed that influenza virus influenced miR-15a expression rising.and then decreased BCL2,CCND1 and P85 target gene. The results induced apoptosis.Bioinformatics analysis of other potential target genes. The finding is that miR-15a can regulate inter-related disease and signal transduction and Significantly enriched pathways are cancer pathways after Application of two calculations to predict the target gene miR-15a, and then Application Biological pathway enrichment analysis and Biocarta pathway database enrichment analysis to analyze Intersection of the result. So the close links may exist between them.Conclusion:the study showing miR-15a take part in biological process of influenza virus inducing apoptosis.With the deepening of the research, we will further understanding the mechanism of miR-15 to influenza virus in order to help the prevention and treatment of influenza virus.