Mouse-specific up-regulation of Ccnb1 expression by miR-199a-5p in keratinocyte

Abstract

MicroRNA (miRNA) are a class of single-stranded, small non-coding RNA that regulate various biological processes, including skin and hair cycle regulation, by modulating the expression of specific genes at the post-transcriptional level. Recently, several studies reported that miRNA directly or indirectly up-regulate target genes. Previously, we performed microarray analysis to identify the target genes of miR-199a-5p in a mouse skin keratinocyte cell line and detected more than 200 genes whose expression was significantly increased by miR-199a-5p overexpression (> 1.5-fold). In this study, we further investigated these genes and found that cyclin B1 (Ccnb1) expression was positively regulated by miR-199a-5p in keratinocyte. Moreover, Ccnb1 expression was inversely correlated with miR-199a-5p expression during the mouse hair cycle. Cell cycle analysis showed that the proportion of cells in S phase was slightly increased, while the proportion of cells in G2/M phase decreased by miR-199-5p. Using luciferase assay, we found that the 3′ untranslated region of Ccnb1 was a direct target of miR-199a-5p. We also found that the regulation of Ccnb1 expression by miR-199a-5p is mouse specific. CCNB1 expression was not affected in the human and monkey cell lines. These results provide a new relationship between Ccnb1 and miR-199a-5p in both mouse keratinocyte and miRNA biology.