MicroRNA-16Affects Cell Motility In Laryngeal Squamous Cell Carcinoma By Targeting Zyxin

Objective:In order to screen out the differentially expressed miRNA in laryngeal squamous cell carcinoma and to understand what the roles they play in tumor process. Methods:The miRNA expressing profiles consisting of210miRNA probes and the mRNA expressing profiles consisting of7267genes of laryngeal carcinoma and normal tissues were compared, and the expression level of interested miRNA was confirmed through real-time PCR. Then, MTT assay, aggregation assay, monolayer wound assay and transwell migration or invasion assay were used to analyze the effect of miR-16on HEp-2cell by blocking miR-16. At the same time, treating HEp-2cell with ASO-16, the target genes of miRNA predicted by bioinformation and cDNA array were confirmed by EGFP reporter system and the level variations of mRNA and protein were detected by real-time PCR and Western blot. At last, we can find out the pathway that miR-16functions in HEp-2cell through overexpression of direct target gene.Results:In this research, we found13miRNAs expressed differentially in laryngeal carcinoma, with6up-regulated and7down-regulated. In mRNA expression profiles,90genes were expressed repeatedly and differentially in3cases of laryngeal carcinoma. Among these genes, up-regulated were29and down-regulated were61. And miR-16-1was the most significantly increased among up-regulated miRNAs. In addition, ASO-16could increase the zyxin expression in both mRNA level and protein level, overexpressed miR-16could decrease the zyxin expression in both mRNA level and protein level, and ASO-16could release the repression of miR-16on EGFP expression intensity in EGFP reporter system and enhance the capability of cell-cell adhesion, inhibit the migration of HEp-2. These results consistented with those obtained from overexpression of zyxin.Conclusion:Microarray was used to screen out the differentially expressed miRNA and mRNA in laryngeal squamous carcinoma. miR-16could reduce the capability of cell-cell adhesion and enhance the migration of cell by targeting zyxin. In this research, it indicated that miRNA took part in the progression of laryngeal carcinoma. And it was helpful for us to understand the mechanism of laryngeal carcinoma biogenesis and development.