The preliminary research of miR-210 promoting human periodontal ligament stem cells the differentiation of angiogenesis / 安徽医科大学学报

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.