Objective To investigate the role of miR-27a in the proliferation, migration and odontoblastic dif-ferentiation of human dental pulp stem cells (hDPSCs), and explore its potential molecular mechanisms. Methods The mRNA and proteins levels of the relevant genes were detected by qRT-PCR and Western blot. hDPSCs proliferation and migration was measured by CCK-8 and Transwell migration assay. Luciferase reporter assay was used to validate the tar-get of miR-27a. Results The over-expression of miR-27a inhibited the proliferation and migration of hDPSCs, and the expression of odontoblastic-related genes (ALP, DMP1 and OCN). The down-regulation of miR-27a increased the prolif-eration and migration of hDPSCs and the expression of ALP, DMP1 and OCN (P<0.05). Luciferase reporter assay showed that Satb2 was a target of miR-27a, and miR-27a negatively regulated the mRNA and protein expression of Satb2 through the 3'' UTR region (P<0.05). The overexpression of Satb2 attenuated the inhibitory effects of miR-27a on the pro-liferation and migration of hDSPCs. Conclusion MiR-27a has suppressive effects on the proliferation and migration of hDPSCs and inhibits the expression of odontoblastic-related genes, which may be related to the regulation of Sabt2 gene.%目的 探讨miR-27a对人牙髓干细胞的增殖、迁移以及成牙本质分化的影响及其潜在的分子作用机制.方法 利用实时荧光定量PCR检测miR-27a以及相关基因的表达水平;分别利用CCK-8实验以及Transwell细胞迁移实验检测人牙髓干细胞的增殖以及迁移能力;通过采用双萤光素酶报告实验验证miR-27a的靶基因;利用Western blot检测相关蛋白的表达水平.结果 MiR-27a过表达可抑制人牙髓干细胞的增殖以及迁移,同时抑制成牙本质向分化相关基因(ALP,DMP1及OCN)的表达(P<0.05);miR-27a的抑制可促进人牙髓干细胞的增殖及迁移,同时增加成牙本质向分化相关基因(ALP,DMP1及OCN)的表达(P<0.05);双萤光素酶报告实验和miR-27a转染实验结果提示miR-27a可以通过作用于Satb2的3''UTR区,负向调控其mRNA及蛋白的表达水平(P<0.05);Satb2的过表达能够拮抗miR-27a过表达对人牙髓干细胞增殖及迁移的抑制(P<0.05).结论 MiR-27a能够抑制人牙髓干细胞的增殖及迁移,同时抑制成牙本质向分化相关基因的表达,这种作用可能与调控Sabt2基因有关.
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