miRNA expression profile during fluid shear stress-induced osteogenic differentiation in MC3T3-E1 cells
miRNA expression profile during fluid shear stress-induced osteogenic differentiation in MC3T3-E1 cells
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DOI：
10.3760/cma.j.issn.0366-6999.20123137
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发表时间：
2013-04-20
影响因子：
6.1
通讯作者：
Ai Hong
中科院分区：
文献类型：
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作者：
Mai Zhi-hui;Peng Zhu-li;Ai Hong
背景：机械应力在维持骨内环境平衡中起着重要作用。目前的假说表明，间质液体流动是骨组织水平应变放大系统的重要组成部分。本研究旨在验证短期和适当的流体剪应力(FSS)有望促进成骨细胞终末分化的假设，并检测在FSS诱导MC3T3-E1细胞成骨分化过程中microRNAs的表达谱。方法采用平行平板流动系统，以12dyn/cm(2)的速度对MC3T3-E1细胞进行1h的FSS处理。FSS治疗后即刻进行细胞骨架免疫组织化学染色和微RNA(MiRNAs)检测。分别于作用后24、48、72h检测成骨基因表达和I型胶原免疫组织化学染色，检测碱性磷酸酶(ALP)活性，第12天进行茜素红染色。结果12dyn/cm(2)FSS作用1h可诱导MC3T3-E1细胞肌动蛋白应激纤维形成和重排，成骨基因表达上调，ALP活性增强，I型胶原合成和分泌增加，结节形成增加，终末分化加快。在成骨分化过程中，FSS诱导的细胞miR-20a、-21、-19b、-34a、-34c、-140和-200b的表达水平明显下调。结论短期、适当的FSS足以促进成骨前细胞的终末分化，一组miRNAs可能参与了FSS诱导的成骨前细胞的分化。
Background Mechanical stress plays an important role in the maintenance of bone homeostasis. Current hypotheses suggest that interstitial fluid flow is an important component of the system by which tissue level strains are amplified in bone. This study aimed to test the hypothesis that the short-term and appropriate fluid shear stress (FSS) is expected to promote the terminal differentiation of pre-osteoblasts and detect the expression profile of microRNAs in the FSS-induced osteogenic differentiation in MC3T3-E1 cells.Methods MC3T3-E1 cells were subjected to 1 hour of FSS at 12 dyn/cm(2) using a parallel plate flow system. After FSS treatment, cytoskeleton immunohistochemical staining and microRNAs (miRNAs) were detected immediately. Osteogenic gene expression and immunohistochemical staining for collagen type I were tested at the 24th hour after treatment, alkaline phosphatase (ALP) activity assay was performed at 24th, 48th, and 72th hours after FSS treatment, and Alizarin Red Staining was checked at day 12.Results One hour of FSS at 12 dyn/cm(2) induced actin stress fiber formation and rearrangement, up-regulated osteogenic gene expression, increased ALP activity, promoted synthesis and secretion of type I collagen, enhanced nodule formation, and promoted terminal differentiation in MC3T3-E1 cells. During osteogenic differentiation, expression levels of miR-20a, -21, -19b, -34a, -34c, -140, and -200b in FSS-induced cells were significantly down-regulated.Conclusion The short-term and appropriate FSS is sufficient to promote terminal differentiation of pre-osteoblasts and a group of miRNAs may be invovled in FSS-induced pre-osteoblast differentiation.