The Regulation Of MiR-1and Mir-184on The Expression Of DAT Gene In SH-SY5Y Cell

Objective:To examine the expression of miR-1、miR-184and Dopamine transporter (DAT) in SH-SY5Y cell, and to explore the effect of miR-1, miR-184on DAT gene expressionMethod:Searched the miRNAs, of which there were the effect on the DAT geneexpression, via the on-line tools such as microRNA.org, miRBase and TargetScan an so on. MiRNAs mimics and inhibitor were chemically synthesized, and transfected into the SH-SY5Y cell. Total RNA and protein were collected after transfecting12h and24h Respectively; miRNAs and DAT mRNA expression in negative control (NC) mimics group, NC inhibitor group, miRNAs" mimics group, miRNAs inhibitor group of SH-SY5Y cell were examined by Real-Time PCR;DAT protein expression in groups of SH-SY5Y cell were detected by westerning blot.Result:(1)Bioinformatics analysis indicated candidate microRNAs which effect the expression of DAT were miR-1and miR-184;(2)The expression of miRNAs:The expression of miR-1between miR-1mimics12h and miR-1mimics24h treatment group cell was significantly higher than that of NC group (Ps<0.001), however there was no significant difference of miR-1expression between in miR-1inhibitor 12h treatment group cell and NC cell, as well as miR-1inhibitor24h treatment group (P>0.05); The expression of miR-184in miR-184mimics24h treatment group cell were significantly different compare with NC group (P<0.01), there was no significant difference of miR-184expression in miR-184inhibitor12h, miR-184mimics12h and and miR-184inhibitor24h treatment groups compared with NC group (Ps>0.05)(3)The expression of DAT mRNAs:the expression of DAT mRNA were significantly different between miR-1mimics12h treatment group cell and NC group (P<0.01), but there were no significant difference of DAT mRNA expression in miR-1mimics24h, miR-1inhibitor12h and miR-1inhibitor24h treatment group cells compare with NC group (Ps>0.05); The expression of DAT mRNA in miR-184mimics24h treatment group cell was significantly different compared with NC group (P<0.01), but not significantly different in miR-184inhibitor12h miR-184mimics12h and miR-184inhibitor24h treatment groups cell (Ps>0.05)(4)The expression of DAT protein:there was significant difference of DAT protein expression between miR-1mimics12h treatment group cells and NC group (P<0.05), however the expression of DAT protein in miR-1mimics24h、miR-1inhibitor12h and miR-1inhibitor24h treatment group cells was significantly different compare with NC group (Ps>0.05); The expression of DAT protein in miR-184mimics24h treatment group cells were significantly different compare with NC group (P<0.05), but not in miR-184inhibitor12h,miR-184mimics12h and miR-184inhibitor24h treatment groups cells (Ps>0.05)Conclusion:MiR-1and miR-184played the role on the regulation of DAT gene expression in SH-SY5Y cell. Over expression of miR-1, miR-184reduced the expression of DAT protein in SH-SY5Y cells.