中国组织工程研究2016，Vol.20Issue(45)：6693-6699,7.DOI:10.3969/j.issn.2095-4344.2016.45.001
miR-195对人骨髓间充质干细胞成骨分化的影响
miR-195 effect on osteogenic differentiation of human bone marrow mesenchymal stem cells
宋鹏 1荆凯 1薛建华 1魏鹏飞1
作者信息
- 1. 河南省中医院，河南省郑州市 450002
- 折叠
摘要
Abstract
BACKGROUND:Previous studies have found that the expression level of miR-195 in differentiated human bone marrow mesenchymal stem cel s (hBMMSCs) is significantly higher than that in undifferentiated hBMMSCs. However, miR-195 effect during this differentiation process and possible mechanism remain unclear. <br> OBJECTIVE:To explore the effect of miR-195 on osteogenic differentiation of hBMMSCs and possible mechanism. <br> METHODS:hBMMSCs were isolated and cultured in vitro. Alkaline phosphatase activity, Runx2, osteopontin and SMAD7 protein expression and miR-195 expression level during osteogenic differentiation of hBMMSCs were determined by alkaline phosphatase kit, western blot and real-time PCR, respectively. miR-195-downexpressed hBMMSCs were constructed by lipofection transfection, and were used to investigate the effect of miR-195 was on osteogenic differentiation of hBMMSCs. Dual luciferase reporter assay was used to identify whether the 3’UTR of SMAD7 mRNA was a binding target of miR-195. In addition, we transfected hBMMSCs with SMAD7 cDNA (pcDNA-SMAD7), and investigated the effect of SMAD7 on osteogenic differentiation of hBMMSCs. <br> RESULTS AND CONCLUSION:The isolated and cultured hBMMSCs had good osteogenic differentiation ability in vitro. Expression level of miR-195 was increased with the increasing of induction time, and the expression level of SMAD7 was reversed. miR-195 promoted osteogenic differentiation of hBMMSCs. Luciferase assay confirmed that miR-195 targeted SMAD7 directly, and overexpression of SMAD7 inhibited the osteogenic differentiation of hBMMSCs. Taken together, miR-195 promotes osteogenic differentiation of hBMMSCs by targeting SMAD7.关键词
干细胞/骨髓干细胞/成骨分化/miR-195/人骨髓间充质干细胞/骨质疏松/SMAD7分类
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宋鹏,荆凯,薛建华,魏鹏飞..miR-195对人骨髓间充质干细胞成骨分化的影响[J].中国组织工程研究,2016,20(45):6693-6699,7.
