Role Of MicroRNAs In Regulating Parkinson's Disease-related Genes

Parkinson's disease (PD) is a common chronic old age, progressive neurodegenerative disease(ND), which pathogenesis have not yet fully clear. MiRNAs are a class of small non-coding RNAs with 17-25 nucleotides in length and believed to be involved in a broad range of biological function by base-pairing with complementary sites of target transcript to negatively regulate gene expression at the posttranscriptional level. It may play an important role in the process and development of PD. Therefore, in-depth study of the regulation mechanism of Parkinson disease-related gene is important for deeply understanding the genetic molecular mechanism of PD, and finding new drugs and understanding the pathogenesis of Parkinson's disease and regulatory networks.In this study, we used a bioinformatics approach to identify conserved miRNAs which can complement with 3'untranslation region of target transcripts involved in Parkinson's disease, these miRNAs were validated by Dual-Luciferase Reporter system. The following is the summarization of our results:1, Using five kinds of miRNA target predict softwares to analyze the genes involved in Parkinson's disease (snca, 1rrk2, park2, pinkl, pitx3), we found numerous miRNA:mRNA base-pairings in the 3'UTR of these genes:98 miRNAs was predicted by miRanda,46 miRNAs was predicted by miRDB,191 miRNAs was predicted by MicroCosm Targets, only 2 miRNAs was predicted by Pictar,97 miRNAs was predicted by TargetScan. Among these 98 base-pairings of miRNAs and their targets predicted by two softwares,15 base-pairings was found in 1rrk2,38 in park2,7 in pitx3,13 in pinkl,10 in snca,15 in uchll.2, Cloning 3'UTR of five mouse Parkinson's disease associated genes' mRNA (snca, Irrk2, park2, pitx3, uchll) and their pairs of 26 miRNAs predicted by the computer.3, After co-transfecting of miRNAs expression constructs and psiCHECK-2-target 3'UTR constructs to mouse eNPC, the activity of Renilla luciferase was measured by GloMaxTM-Multi+ and firefly luciferase was used as internal control to normalize Renilla luciferase data. The resultes showed that: miR-153, miR-350, miR-141 can down regulate snca; miR-27a/27b, miR-29a, miR-466k can down regulate park2; miR-183, miR-133a/b, miR-30b, miR-211 can down regulate pitx3; miR-92a, miR-466g, miR-9, miR-20a, miR-17 can down regulate uchll; miR-30a, miR-30b, miR-9, miR-30e, miR-384-5p, miR-467b can down regulate 1rrk2, suggesting that miRNAs are involved in the genes of Pakinson's disease.Our works laid the foundation of uncovering of the function and mechanism about miRNA in the Pakinson's disease, and make a new thinking about the development of therapy and the new medicine of Pakinson's disease.