MiR-203 regulates neuroblastoma cell migration by targeting CAMTA1

Objective To explore the expression level of miR-203 in neuroblastoma tissue and elucidate its mechanism in neuroblastoma cell migration process. Methods In neuroblastoma cells transfected with miR-203 inhibitor, the impact of miR-203 upon migratory capacity was studied through Transwell assay. The bioinformatics was used to predict the target of miR-203. And the verification methods included luciferase reporter assay, real-time quantitative PCR and Western blot. The target gene CAMTA1 of miR-203 was confirmed. SiRNA interference technology was adopted to suppress the expression of target gene CAMTA1. Through co-transfections of miR-203 inhibitor and CAMTA1 siRNA, we determined whether miR-203 affected migration directly by targeting CAMTA1. And the expression of miR-203 was detected in metastatic and primary neuroblastoma specimens. Results Compared with ASO control, the numbers of migratory neuroblastoma cells SH- SY-SY and SK-N-SH decreased by 41.34% and 37. 860% after transfection with miR-203 ASO （P〈 0. 05）. MiR-203 could directly target CAMTA1 mRNA wild type 3＂UTR. After transfection with miR-203 ASO, the endogenous expression levels of CAMTA1 increased by 2. 13 and 1.78 folds in SH- SY-5Y and SK-N-SH cells compared with ASO control （P〈0. 05, P〈0. 05）. After transfection with CAMTA1 cDNA plasmid, the number of migration in SH-SY-fY and SK-N-SH decreased by 25.27% and 19. 480% respectively （P〈0. 05）. After co-transfections with miR-203 inhibitor and CAMTA1 siRNA, the number of migratory SH-SY5Y cell increased by 19. 37% compared with miR-203 inhibitor （P〈0. 05） and its number showed no significance compared with inhibitor control （P〈 0. 05）. MiR-203 had a higher expression level in metastatic neuroblastoma specimen than primary neuroblastoma specimen （P〈0. 05）. Conclusions MiR-203 is correlated with migratory capacity and it promotes the migration of neuroblastoma cell directly by targeting CAMTA1. Key words: Neuroblastoma;  RNA, small interfering;  Cell migration assays