Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3
Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3
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失调的 miR-155 通过靶向 FADD 和 caspase-3 促进人椎间盘退变中 Fas 介导的细胞凋亡
DOI：
10.1002/path.2931
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发表时间：
2011-10-01
影响因子：
7.3
通讯作者：
Luo, Zhuo-Jing
中科院分区：
文献类型：
--
作者：
Wang, Hai-Qiang;Yu, Xiao-Dong;Luo, Zhuo-Jing
细胞凋亡在椎间盘退变（IDD）发病机制中的作用仍然是个谜。越来越多的证据表明，细胞凋亡机制是由miRNA调控的。我们推测miRNAs可能参与IDD的细胞凋亡。我们发现29种miRNAs在退变髓核中差异表达，miR-155在退变髓核中下调。使用实时PCR进一步验证miR-155的失调（0.56倍，p < 0.05）。生物信息学靶点预测将FADD和caspase-3确定为miR-155的推定靶点。此外，miR-155通过直接靶向FADD和caspase-3的3 '-UTR来抑制它们的表达，这通过miR-155结合位点的突变来消除。通过用慢病毒pre-miR-155转染在人NP细胞中miR-155的体外上调导致FADD和半胱天冬酶-3的抑制;而用慢病毒antigomiR-155敲低miR-155导致FADD和半胱天冬酶-3的过表达。此外，Fas介导的细胞凋亡在拮抗miR-155时增加，而在人NP细胞中使用pre-miR-155时减少。此外，我们提出了直接的证据，NP细胞凋亡IDD组织中使用透射电子显微镜分析。此外，原位杂交（ISH）和免疫组织化学（IHC）的组合显示，miR-155在人NP细胞的细胞质中表达，与FADD和caspase-3呈负相关。总之，这是第一个研究IDD在细胞凋亡和miRNA方面的潜在机制。此外，caspase-3被鉴定为miR-155的新靶点。我们的研究结果表明，去调控的miR-155通过靶向FADD和caspase-3促进Fas介导的人IDD细胞凋亡，暗示miR-155在IDD中的病因学和治疗作用。版权所有（C）2011大不列颠和爱尔兰病理学会。由John Wiley & Sons有限公司出版
The role of apoptosis in the pathogenesis of intervertebral disc degeneration (IDD) remains enigmatic. Accumulating evidence has shown that the apoptotic machinery is regulated by miRNAs. We hypothesized that miRNAs might contribute to apoptosis in IDD. We have found that 29 miRNAs were differentially expressed and miR-155 was down-regulated in degenerative nucleus pulposus (NP). The deregulation of miR-155 was further verified using real-time PCR (0.56 fold, p < 0.05). Bioinformatics target prediction identified FADD and caspase-3 as putative targets of miR-155. Furthermore, miR-155 inhibited FADD and caspase-3 expression by directly targeting their 3'-UTRs, which was abolished by mutation of the miR-155 binding sites. In vitro up-regulation of miR-155 in human NP cells by transfection with lentiviral pre-miR-155 resulted in repression of FADD and caspase-3; whereas knockdown of miR-155 with lentiviral antigomiR-155 led to over-expression of FADD and caspase-3. Also, Fas-mediated apoptosis was increased when antagonizing miR-155 and decreased when using pre-miR-155 in human NP cells. In addition, we presented direct evidence of NP cells undergoing apoptosis in IDD tissues using transmission electron microscopy analysis. Moreover, a combination of in situ hybridization (ISH) and immunohistochemistry (IHC) revealed that miR-155 expressed in the cytoplasm of human NP cells with reverse correlation with FADD and caspase-3. In summary, this is the first study addressing the underlying mechanisms of IDD in terms of apoptosis and miRNAs. Furthermore, caspase-3 is identified as a novel target of miR-155. Our results suggest that deregulated miR-155 promotes Fas-mediated apoptosis in human IDD by targeting FADD and caspase-3, implicating an aetiological and therapeutic role of miR-155 in IDD. Copyright (C) 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.