The Function Of MiR-25 In Invasion And Metastasis Of Esophageal Squamous Carcinoma And The Bioinformatics Analysis Of MiR-25

Objective: To investigate the role mi R-25 plays in invasion and metastasis of esophageal squmous carcinoma, and analyse its bioinformatic messages.Methods: To inhibit and overexpress mi R-25 by transfecting mi RNA inhibitor and mi RNA mimic into human esophageal carcinoma cells KYSE-150, EC109,then study the function mi R-25 plays in different groups of cells through migration assay, invasion assay and wound healing assay. After that, mi R-25 targeted genes were predicted via Target Scan, Pic Tar, mi Randa and mi RTarbase, the function of the target genes were predicted via Gene Ontology(GO) analysis while the enriched KEGG pathway analyses of mi R-25 targeted genens were performed by bioinformatics tool DAVID 6.7 and mi RNA target gene prediction database mirfocus 3.0. Then protein-protein interaction networks of the targeted gene were produced by STRING.Results: The expression level of mi R-25 was significantly different in three groups of cells(P <0.05), then the overexpression, high expression and low expression model of mi R-25 in esophageal squamous cell carcinoma cells have been down.(1)KYSE-150: Transwell cell invasion assay showed that the number of cells invated into the endercell of the group mimic was significently more than that of the group inhibitor(p<0.01), and the number of cells invated into the undercell of the group inhibitor was significantly less than that of the group negative control(p<0.01); Transwell cell migration assay showed that the number of cells migrated into the undercell of the group mimic was significantly more than that of the group inhibitor(p<0.01), and the number of cells migrated into the endercell of the group inhibitor was significently less than that of the group negative control(p<0.01); cells transfected with mimic reagent closed the scratchwounds more quickly than cells that were transfected with inhibitor reagent or negative control reagent which meant that migration of group mimic was significantly enhanced compared with group inhibitor( p<0.01),and migration of group inhibitor was significantly decreased compared with group negative control(p<0.01)after 12 h, the differences were significant as well after 24h(p<0.05).(2) EC109: the results of group mimic of Transwell cell migration and invasion assay are the same as KYSE-150, but the results of group inhibitor are different, the numbesr of cells of group inhibitor are close to the number of cells of negative control(p>0.05).(3)four online bioinformatics databases(Target Scan6.1, Pic Tar, mi Randa, and mi RTarbase) to select plausible targets and validated targets of mi R-25, and finally obtained 54 target genes : ZNF512B、ERC2、MIER3、DMXL1、EXOC5、PRKCE、PCDH11Y、PDS5B、LHFPL2、COL1A2, et al. Gene Ontology and DAVID 6.7. were used for the next analysis aiming at the 54 target genes, the results showed that the predicted target genes mainly enriched in regulation of cellular metabolic process, positive regulation of cellular metabolic process, regulation of primary metabolic process, et al(p<0.001), and they mainly enriched in protein binding, ubiquition protein ligase binding, small conjugating protein ligasse binding, etal(p<0.001),the only significant cellular component was nucleus with a p value of 7.904е-03. Enriched KEGG pathways by DAVID 6.7 displayed in KEGG pathway database showed that, the predicted target genes of mi R-25 were significantly enriched in the p53 signaling pathway(p53 pathway) and tumor signaling pathways(pathways in cancer), both of them are tumor-related signaling pathway(p <0.05). STRING showed that the protein interaction existed in total 26 protein targeted by the predicted genes, which together formed the target gene interaction network. TP53, KAT2 B and MDM2 whose connections were very close in the network may incoded important downstream target proteins.Conclusion: mi R-25 promotes invasion and metastasis of esophageal squmous cancer; the influence of inhibiting the expression of mi R-25 in ESCC cell invasion and metastasis may be relevant with cell differentiation: lower the degree of differentiation, more significant the inhibition. The predicted target genes TP53, MDM2 and KAT2 B may be downstream target genes of mi R-25, the proteins network system interact with other relevant roles in cell metabolism, protein binding and tumor-related pathway, resulting in tumor formation and development.