[Differential expression profile of microRNAs between human bone marrow mesenchymal stem cells and sweat gland-like cells].

ABSTRACT

OBJECTIVE: To explore the differential microRNAs (miRNAs) expression profiles between human bone marrow mesenchymal stem cells (hBM-MSCs) and sweat gland-like cells. METHODS: The hBM-MSCs were induced to differentiate into osteogenic and adipogenic cells by related osteogenic differentiation basal medium and adipogenic differentiation basal medium respectively. And the results were assayed by oil red O and alizarin red stains respectively. When reaching 70%-80% confluence, primarily cultured sweat gland cells were heat-shocked and co-cultured with hBM-MSCs by a Transwell plate to induce the differentiation of hBM-MSCs. After 10 days, the total RNAs of hBM-MSCs and sweat gland-like cells were extracted, purified, labeled, hybridized and analyzed for predicting miRNAs target genes separately. The microarray results were confirmed by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: There were 68 miRNAs of >double differentiation on expression level between hBM-MSCs and sweat gland-like cells. And 19 miRNAs were up-regulated and 49 miRNAs down-regulated. Moreover, there were 13 miRNAs with >5 folds of differential expression level between hBM-MSCs and sweat gland-like cells, including 5 up-regulated miRNAs (miRNA-132-3p, -4467, -4484, -146a-5p and -6126) and 8 down-regulated miRNAs (miRNA-708-5p, -138-5p, -6812-5p, -138-1-3p, -1281, -3157-3p, -4298 and -4459). There were 18 miRNAs related to regulation of the signaling pathways of sweat gland development, including nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase/extracellular regulated protein kinase (MAPK/Erk), Wnt/beta-Catenin (Wnt/ß-Catenin) and ectodysplasin-A1/ectodysplasin-A1 receptor (EDA/EDAR). The results of RT-PCR on miRNA-146a-5p and miRNA-6812-5p had a high concordance with the results of microarray. CONCLUSIONS: There are obvious differentiation miRNAs expression profiles between hBM-MSCs and sweat gland-like cells. And 13 miRNAs >5 folds of differential expression and 18 miRNAs related to sweat gland development may play essential roles in regulated differentiation of hBM-MSCs into sweat gland-like cells.