标题：MiR-27a对人牙髓干细胞增殖、迁移及成牙本质分化的作用

      作者：谢双文 1，蒋才丽 2，王伯钧 2    (1.桂林医学院口腔科，广西 桂林 541001；2.广西科技大学第一附属医院口腔科，广西 柳州 545002)

      卷次： 2018年29卷11期

      【摘要】 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。
      【关键词】 MiR-27a；人牙髓干细胞；Satb2；增殖；迁移；成牙本质向分化相关基因
      【中图分类号】 R781.3 【文献标识码】 A 【文章编号】 1003—6350（2018）11—1481—05

Effects of miR-27a on the proliferation, migration and odontoblastic differentiation of human dental pulp stemcells.
XIE Shuang-wen 1, JIANG Cai-li 2, WANG Bo-jun 2. 1. Department of Stomatology, Guilin Medical University, Guilin541001, Guangxi, CHINA; 2. Department of Stomatology, the First Affiliated Hospital of Guangxi University of Science andTechnology, Liuzhou 545002, Guangxi, CHINA
【Abstract】 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 ThemRNA and proteins levels of the relevant genes were detected by qRT-PCR and Western blot. hDPSCs proliferation andmigration 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 theexpression 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 assayshowed that Satb2 was a target of miR-27a, and miR-27a negatively regulated the mRNA and protein expression of Satb2through 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 ofhDPSCs and inhibits the expression of odontoblastic-related genes, which may be related to the regulation of Sabt2 gene.
      【Key words】 MiR-27a; Human dental pulp stem cells (hDPSCs); Satb2; Proliferation; Migration; Odontoblas-tic-related genes·论 著·doi:10.3969/j.issn.1003-6350.2018.11.001