安徽医科大学学报2015，Vol.50Issue(10)：1381-1385,5.
miR-210 促进人牙周膜干细胞血管向分化作用的初步研究
The preliminary research of miR-210 promoting human periodontal ligament stem cells the differentiation of angiogenesis
摘要
Abstract
Objective To construct the lentiviral vector Lenti-miR-210-Luciferase, and to detect angiogenic factors HIF-1α and VEGF expression in hPDLSCs transduced by Lenti-miR-210-Luciferase. Methods hPDLSCs were iso-lated and cultured, and according to human miR-210 gene sequence(NC_000011. 9), its primer was designed and amplified through PCR. The PCR products of the target gene were connected to the vector pLVX-EGFP-3FLAG-EF1-Luc. To identify the plasmid, target gene PCR product and the purpose vector were digested by EcoRⅠand XbaⅠ. Lenti-miR-210-Luciferase ( the control group was Lenti-LacZ-Luciferase) was constructed using the LR re-combination system. After hPDLSCs was transduced by Lenti-miR-210-Luciferase, the analysis of HIF-1α and VEGF expression was done with qPCR and the immunohistochemistry examination. Results The results of plasmid sequencing and digestion confirmed that the vector Lenti-miR-210-Luciferase was successfully constructed. After Lenti-miR-210-Luciferase was transduced to hPDLSCs on 0, 1, 4, 7 and 14 d, the results of qPCR showed that the over-expression of HIF-1αand VEGF was detected on 4 d, and continued until 21 d. Immunohistochemical results showed that after hPDLSCs were transduced by Lenti-miR-210-Luciferase, hPDLSCs were positive for HIF-1α and VEGF antibody, and the control group was negative. Conclusion The Lenti-miR-210-Luciferase is successfully constructed, and miR-210 can promote hPDLSCs the differentiation of angiogenesis.关键词
牙周膜干细胞/miR-210/低氧诱导因子-1α/血管内皮生长因子/慢病毒载体Key words
periodontal ligament stem cells/miR-210/hypoxia inducible factor-1α/vascular endothelial growth factor/lentiviral vector分类
生物科学引用本文复制引用
古月,王银龙..miR-210 促进人牙周膜干细胞血管向分化作用的初步研究[J].安徽医科大学学报,2015,50(10):1381-1385,5.基金项目
安徽省科技厅重点课题(编号:10021303003) （编号:10021303003）
