LncRNA LINC00311 Promotes the Proliferation and Differentiation of Osteoclasts in Osteoporotic Rats Through the Notch Signaling Pathway by Targeting DLL3 (Retracted article. See vol. 55, pg. 240, 2021)
LncRNA LINC00311 Promotes the Proliferation and Differentiation of Osteoclasts in Osteoporotic Rats Through the Notch Signaling Pathway by Targeting DLL3 (Retracted article. See vol. 55, pg. 240, 2021)
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DOI：
10.1159/000491539
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发表时间：
2018-01-01
影响因子：
--
通讯作者：
Cui, Zhi-Qiang
中科院分区：
文献类型：
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作者：
Wang, Yu;Luo, Tian-Bao;Cui, Zhi-Qiang
背景/目的：骨质疏松症是一种以骨密度丢失为特征的常见疾病。先前的证据强调了microRNA作为疾病潜在治疗工具所发挥的作用。目前，长链非编码RNA（lncRNA）对骨质疏松症进展的影响在很大程度上仍不清楚。因此，结合一系列生物信息学方法，对已报道在骨质疏松症中高度表达的LINC 00311和参与Notch信号传导途径的δ样3（DLL 3）之间的靶向关系进行了研究。通过卵巢切除术（OVX）建立骨质疏松大鼠模型，以评估DLL 3结合LINC 00311通过Notch信号通路对破骨细胞的影响。研究方法：从骨质疏松大鼠中提取破骨细胞，并用LINC 00311-载体、shRNA-LINC 00311、Notch激活剂或Notch激活剂和LINC 00311-载体的组合转染。采用Western blotting和RT-qPCR技术检测LINC 00311、DLL 3、Notch 1、Notch 2、Jagged 1、Hes-1和TRAP在组织和细胞中的表达水平，MTT法检测细胞活性。流式细胞仪检测细胞周期和凋亡率。将造模成功的大鼠分为OVX siRNA组、OVX LINC 00311组和OVX对照组，观察LINC 00311对破骨细胞增殖和分化的影响。结果如下：用LINC 00311-载体转染的细胞表现出Notch 2和TRPA的表达水平增加以及细胞活性增加，同时观察到DLL 3、Notch 1、Jagged 1和Hes-1的表达水平降低，沿着细胞凋亡率降低。在用shRNA-LINC 00311处理的细胞中观察到这些参数的相反趋势。当Notch信号通路被激活时，进行了关键的观察，即细胞活性降低，而凋亡率增加。与OVX对照组相比，OVX siRNA组LINC 00311、Notch 2和TRAP的表达水平以及TRAP的阳性表达率均降低，DLL 3、Jagged 1和Notch 1的表达水平升高。与假手术组相比，OVX对照组和OVX LINC 00311组LINC 00311表达增加，Notch 2和TRAP表达增加，DLL 3、Jagged 1和Notch 1表达减少。结论：综上所述，本研究的关键发现表明，LINC 00311通过抑制DLL 3表达来调节Notch信号通路，从而诱导破骨细胞增殖并抑制其凋亡，最终证明LINC 00311及其靶基因DLL 3可能是骨质疏松症的独立因素。(C)2018作者（S）由S发布。Karger AG，巴塞尔
Background/Aims: Osteoporosis is a commonly occurring condition marked by a loss of bone density. Previous evidence has highlighted the roles played by microRNAs as potential treatment tools for the disease. At present, the influence of long non-coding RNAs (lncRNAs) on the progression of osteoporosis remains largely unclear. Thus, an investigation was conducted into the target relationship between LINC00311, which has been reported to be highly expressed in osteoporosis, and delta-like 3 (DLL3), which is involved in the Notch signaling pathway, in connection with a series of bioinformatic methods. An osteoporotic rat model was established by means of ovariectomy (OVX) to evaluate the influence exerted by DLL3-binding LINC00311 on osteoclasts through the Notch signaling pathway. Methods: Osteoclasts were extracted from osteoporotic rats and transfected with the LINC00311-vector, shRNA-LINC00311, Notch activator, or a combination of the Notch activator and LINC00311-vector. Western blotting and RT-qPCR techniques were applied to determine the expression levels of LINC00311, DLL3, Notch1, Notch2, Jagged1, Hes-1 and TRAP in tissues and cells, while cell activity was detected by MTT assay. The cell cycle as well as the rate of apoptosis was detected by flow cytometry. The successfully established osteoporotic rats were designated into the OVX-siRNA, OVX-LINC00311 and OVX-control groups to observe the effects of LINC00311 on the proliferation and differentiation of osteoclasts. Results: Cells transfected with the LINC00311-vector exhibited increased expression levels of Notch2 and TRPA as well as increased cell activity, while decreased expression levels of DLL3, Notch1, Jagged1 and Hes-1, along with a decreased cell apoptosis rate, were observed. The opposite tendencies of these parameters were observed in the cells treated with shRNA-LINC00311. A key observation was made when the Notch signaling pathway was activated, in that the cell activity was decreased while the rate of apoptosis increased. In comparison with the OVX-control group, the expression levels of LINC00311, Notch2 and TRAP as well as the positive expression rate of TRAP all exhibited reductions, while those of DLL3, Jagged1 and Notch1 were elevated in the OVX-siRNA group. Compared with those in the sham group, in the OVX-control and OVX-LINC00311 groups, LINC00311 and the expression levels of Notch2 and TRAP were increased; however, decreased levels of DLL3, Jagged1 and Notch1 were noted. Conclusions: Taken together, the key findings of the present study suggest that LINC00311 induces proliferation and inhibits apoptosis of osteoclasts via the regulation of the Notch signaling pathway by inhibiting DLL3 expression, ultimately demonstrating that LINC00311 and its target gene DLL3 may serve as independent factors in cases of osteoporosis. (C) 2018 The Author(s) Published by S. Karger AG, Basel