miR-103对牙髓干细胞增殖及其向成牙本质细胞分化的影响

The effects of miR-103 on the proliferation and differentiation of human dental pulp stem cells

目的:探讨miR-103对牙髓干细胞向成牙本质细胞分化的影响。方法:从即刻拔除的成人第三磨牙中分离培养人牙髓干细胞,将培养的细胞分为对照组、miRNA阴性对照组、miR-103过表达组和miR-103降低表达组,利用CCK-8法检测人牙髓细胞的增殖能力变化,利用q-PCR和Western Bolt技术检测人牙髓细胞中Runt相关转录因子2(Runx2)、成骨细胞特异性转录因子(Osx)和碱性磷酸酶(ALP)等分化指标的含量变化,用茜素红染色法检测人牙髓细胞的矿化能力。结果:过表达miR-103后人牙髓细胞的增殖能力下降(P<0.05),分化指标含量减少(P<0.05),矿化结节数量减少(P<0.05);降低miR-103后人牙髓细胞的增殖能力增强(P<0.05),分化指标含量增多(P<0.05),矿化结节数量增多(P<0.05)。结论:miR-103抑制牙髓干细胞的增殖,且抑制其向成牙本质细胞的分化。

Objective:To investigate the effects of miR-103 on the proliferation and differentiation of dental pulp stem cells(DPSCs).Methods:Human DPSCs were isolated from extracted adult third molars immediately after extraction.The cultured cells were divided into control group,miRNA negative control group(miR-NC),miR-103 overexpression group(miR-mimic)and miR-103 knock down group(anti-miR).Transfection technology was used to overexpress and reduce the content of miR-103 in DPSCs.CCK-8 method was used to detect the proliferation of DPSCs.q-PCR and Western bolt were used to detect the content of runt-related transcription factor 2(Runx2),Osterix(Osx)and alkaline phosphatase(ALP)in DPSCs.The mineralization ability of DPSCs was detected by alizarin red staining.Results:Over expression of miR-103 promoted the proliferation of DPSCs(P<0.05),decreased the content of Runx2,Osx and ALP,decreased the number of mineralized nodules of DPSCs(P<0.05).Knocking down miR-103 increased the proliferation of DPSCs(P<0.05),increased the content of Runx2,Osx and ALP and increased in the number of mineralized nodules of DPSCs(P<0.05).Conclusion:miR-103 inhibits the proliferation and differentiation of DPSCs into odontoblasts.