标题：miR-217在hADSCs增殖与成骨分化中的作用及其机制研究

      作者：韦宗勇 1，肖展宏 1，黎裕明 2    南宁市第一人民医院康复医学科 1、骨科 2，广西 南宁 530001

      卷次： 2020年31卷13期

      【摘要】 目的 探索microRNA-217 (miR-217)在调控人脂肪组织源性间充质干细胞(hADSCs)增殖与成骨分化中的作用机制。方法 分离、纯化培养人hADSCs，体外诱导成骨分化，基因芯片检测hADSCs向成骨细胞分化过程中microRNA表达。向hADSCs转染miR-217模拟物或miR-217抑制剂，CCK8检测细胞增殖，实时定量PCR检测成骨细胞特异转录因子(Osterix)、骨钙蛋白(OCN)、Runt相关转录因子 2 (Runx2)、碱性磷酸酶(ALP)的表达变化。结果 基因芯片显示hADSCs成骨细胞分化过程中多个microRNA表达上调，其中miR-412-5p、miR-19a-3p、miR-217升高趋势最明显，qPCR显示在hADSCs向成骨细胞分化过程中miR-217升高最显著，差异有统计学意义(P<0.05)；miR-217 模拟物能够促进 hADSCs细胞增殖、而miR-217 抑制剂能抑制增殖；miR-217模拟物促进hADSCs内Osterix、OCN、Runx2、ALP mRNA的表达，相反miR-217抑制剂显著抑制上述基因表达，差异均有统计学意义(P<0.05)。结论 miR-217能促进hADSCs增殖，通过正向调控成骨相关基因从而促进hADSCs向成骨细胞分化。
      【关键词】 miR-217；人脂肪组织源性间充质干细胞；细胞增殖；成骨分化；作用机制
      【中图分类号】 R336 【文献标识码】 A 【文章编号】 1003—6350（2020）13—1633—05

Role and mechanism of miR-217 in the proliferation and osteogenic differentiation of hADSCs.
WEI Zong-yong 1,XIAO Zhan-hong 1, NI Yu-ming 2. Department of Rehabilitation Medicine 1, Department of Orthopaedics 2, Nanning FirstPeople's Hospital, Nanning 530001, Guangxi, CHINA
【Abstract】 Objective To investigate the molecular mechanisms of microRNA-217 (miR-17) in the prolifera-tion and osteogenic differentiation of human adipose-derived mesenchymal stem cells (hADSCs). Methods hADSCswere isolated, purified, and induced in vitro for osteogenic differentiation. Gene chip was used to detect the expression ofmicroRNA during the osteogenic differentiation of hADSCs. miR-217 mimic or inhibitor was transfected into hADSCs,and the cell proliferation was detected by CCK8. The expression of osteoblast specific transcription factor (Osterix), os-teocalcin (OCN), Runt-related transcription factor-2 (Runx2), alkaline phosphatase (ALP) was detected by real-timequantitative PCR. Results Gene chip showed the expression of miR-412-5p, miR-19a-3p, miR-217 were unregulat-ed in the process of hADSCs osteogenic differentiation. qPCR showed that miR-217 was elevated most significantlyduring osteogenic differentiation of hADSCs (P<0.05). miR-217 mimic promoted the proliferation of hADSCs cells,while miR-217 inhibitor suppressed the proliferation. The expression of Osterix, ALP, OCN, Runx2 mRNA were in-duced by miR-217 mimic, and were inhibited by miR-217 inhibitor. Conclusion MiR-217 can promote the prolifer-ation of hADSCs, and promote the differentiation of hADSCs into osteoblasts by positively regulating osteogenic relat-ed genes.
      【Key words】 miR-217; hADSCs; Cell proliferation; Osteogenic differentiation; Mechanism