miR-98-TXLNG1 (FIAT)/Sp7 function loop mediates osteoblast mineralization.
miR-98-TXLNG1 (FIAT)/Sp7 function loop mediates osteoblast mineralization.
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DOI：
10.26355/eurrev_201806_15246
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发表时间：
2018-06
影响因子：
3.3
通讯作者：
L. Yang;L-Y Huang;L-B Li;Z. Tang;K. Chen;L.-P. Chen;H-Y Luo;J. Luo;X.-L. Zhao
中科院分区：
文献类型：
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作者：
L. Yang;L-Y Huang;L-B Li;Z. Tang;K. Chen;L.-P. Chen;H-Y Luo;J. Luo;X.-L. Zhao
目的探讨microRNAs （miRNAs）在成骨细胞矿化中的作用及其机制。材料与方法采用实时聚合酶链反应（Real-time polymerase chain reaction， PCR）、Northern Blot和Western Blot检测调节因子的表达模式。我们通过过表达和下调实验来研究miR-98的作用以及调节因子之间的相互作用。利用生物信息学计算和荧光素酶报告基因实验对靶基因进行验证。电泳迁移率转移实验（EMSA）、染色质免疫沉淀（CHIP）和启动子荧光素酶报告子实验证实了miR-98的调控因子和启动子之间的关系。结果MiR-98在成骨细胞矿化过程中表达上调。过表达miR-98促进成骨细胞矿化。因子抑制激活转录因子4 （ATF4）介导的转录（FIAT）是成骨细胞分化的负调节因子，被证实是miR-98的靶标。作为成骨细胞矿化的激励因子，Sp7转录因子7 （Sp7）通过在启动子区域的组合促进miR-98的转录。我们的研究表明，miR-98是成骨细胞矿化的重要调节因子，miR-98通过一个新的miR-98- fiat /Sp7调节环发挥其功能。这为mirna在成骨细胞矿化中的作用提供了新的见解。
OBJECTIVE To investigate the role of microRNAs (miRNAs) and its mechanism in osteoblast mineralization. MATERIALS AND METHODS Real-time polymerase chain reaction (PCR), Northern Blot, and Western Blot were used to identify the expression mode of regulators. Overexpression and down-regulation experiments were carried out to study the role of miR-98 and interactions between regulators. Bioinformatics calculation and luciferase reporter assay were used to prove the target gene. Electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (CHIP), and promoter luciferase reporter assay confirmed the relationship between the regulator and the promoter of miR-98. RESULTS MiR-98 was up-regulated during osteoblast mineralization. Overexpression of miR-98 promoted osteoblast mineralization. Factor inhibiting activating transcription factor 4 (ATF4)-mediated transcription (FIAT), a negative regulator of osteoblast differentiation, was confirmed to be a target of miR-98. As a motivator in osteoblast mineralization, Sp7 transcription factor 7 (Sp7) promoted miR-98 transcription by a combination on the promoter region. CONCLUSIONS Our study showed that miR-98 was an important regulator in osteoblast mineralization and miR-98 carried out its function through a novel miR-98-FIAT/Sp7 regulatory loop. It provides new insights into the roles of miRNAs in osteoblast mineralization.