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miR-let-7b Lentiviral Vector Promote the Differentiation of Bone Marrow Mesenchymal Stem Cells into Nerve Cells
Zhou Xueying1, Li Shuangyue1, Qu Shuxian2, Qu Yanhui3, Shao Ying4, Sun Jingsong4*, Piao Fengyuan1*
1The Department of Public Health, Dalian Medical University, Dalian 116044, China; 2Tumor Stem Cell Research Institute of Dalian Medical University, Dalian 116044, China; 3The Second Affiliated Hospital of Dalian Medical University, Dalian 116011, China; 4The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China
Abstract: MicroRNAs play an important role in the process of growth and senescence of living organisms， and participate in the important biological processes of MSCs， including proliferation， differentiation， signal transduction and death. The effect of miR-let-7 family members on the differentiation of bone marrow mesenchymal stem cells into neurons can promote stem cell transplantation. To investigate the role of miR-let-7b in promoting the differentiation of rat bone marrow mesenchymal stem cells into nerve cells， the rat miR-let-7b lentiviral vector and Anti-rno-miR-let-7b Inhibitor vector were transfected into rat bone marrow mesenchymal stem cells in vitro. The experiment was divided into control group (MiR-let-7b lentiviral) and miR-let-7b-group (transfected with Anti-rno-miR-let-7b Inhibitor). Furthermore， the effects of all-trans retinoic acid (RA)， basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were used to induce bone marrow. The expression of miR-let-7b in three groups of cells was compared by Real-time quantitative PCR (RT-qPCR). The expression of NSE was detected by immunocytochemical staining. The expression of MAP-2 mRNA was detected by RT-qPCR. The expression of CD90 and CD44 was more than 90% and the expression of CD45 was less than 2%. The results showed that the cells were MSCs. RT-qPCR results showed that the miR-let-7b expression level in the miR-let- 7b+ group was higher than that in the control group， and miR-let-7 was hardly detected in the miR-let-7b- group， suggesting that miR-let-7b vector and miR-let-7b Inhibitor vector were successfully transfected with MSCs. The expression of SIM312， Gap43， MBP and NSE protein in miR-let-7b+ group was significantly higher than these in control group (P<0.05). Meanwhile， The expression of SIM312， Gap43， MBP and NSE protein had significantly differences in the groups with or without miR-let-7 (P<0.05). The results suggest that miR-let-7b can promote the differentiation of MSCs into neurons and regulate the differentiation rate of MSCs by controlling the level of miRlet- 7b.
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