lncRNA SNHG1 受 SP1 诱导通过海绵吸附 miR-181c-5p 并调节 SFRP1/Wnt 信号通路来调节骨重塑和血管生成。
lncRNA SNHG1 induced by SP1 regulates bone remodeling and angiogenesis via sponging miR-181c-5p and modulating SFRP1/Wnt signaling pathway.
机构信息
Department of Orthopedics, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo, 315000, Zhejiang Province, China.
Ningbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, No. 41 Xibei Street, Ningbo, 315000, Zhejiang Province, China.
出版信息
Mol Med. 2021 Nov 3;27(1):141. doi: 10.1186/s10020-021-00392-2.
BACKGROUND
We aimed to investigate the functions and underlying mechanism of lncRNA SNHG1 in bone differentiation and angiogenesis in the development of osteoporosis.
METHODS
The differential gene or proteins expressions were measured by qPCR or western blot assays, respectively. The targeted relationships among molecular were confirmed through luciferase reporter, RIP and ChIP assays, respectively. Alkaline phosphatase (ALP), alizarin red S (ARS) and TRAP staining were performed to measure the osteoblast/osteoclast differentiation of BMSCs. The viability, migration and angiogenesis in BM-EPCs were validated by CCK-8, clone formation, transwell and tube formation assays, respectively. Western blot and immunofluorescence detected the cytosolic/nuclear localization of β-catenin. Ovariectomized (OVX) mice were established to confirm the findings in vitro.
RESULTS
SNHG1 was enhanced and miR-181c-5p was decreased in serum and femoral tissue from OVX mice. SNHG1 directly inhibited miR-181c-5p to activate Wnt3a/β-catenin signaling by upregulating SFRP1. In addition, knockdown of SNHG1 promoted the osteogenic differentiation of BMSCs by increasing miR-181c-5p. In contrast, SNHG1 overexpression advanced the osteoclast differentiation of BMSCs and inhibited the angiogenesis of BM-EPCs, whereas these effects were all reversed by miR-181c-5p overexpression. In vivo experiments indicated that SNHG1 silencing alleviated osteoporosis through stimulating osteoblastogenesis and inhibiting osteoclastogenesis by modulating miR-181c-5p. Importantly, SNHG1 could be induced by SP1 in BMSCs.
CONCLUSIONS
Collectively, SP1-induced SNHG1 modulated SFRP1/Wnt/β-catenin signaling pathway via sponging miR-181c-5p, thereby inhibiting osteoblast differentiation and angiogenesis while promoting osteoclast formation. Further, SNHG1 silence might provide a potential treatment for osteoporosis.
背景
我们旨在研究 lncRNA SNHG1 在骨质疏松症发展过程中的骨分化和血管生成中的功能和潜在机制。
方法
通过 qPCR 或 Western blot 测定差异基因或蛋白表达。通过荧光素酶报告、RIP 和 ChIP 测定分别验证分子之间的靶向关系。通过碱性磷酸酶（ALP）、茜素红 S（ARS）和 TRAP 染色测定 BMSCs 的成骨/破骨细胞分化。通过 CCK-8、克隆形成、transwell 和管形成测定分别验证 BM-EPCs 的活力、迁移和血管生成。Western blot 和免疫荧光检测 β-连环蛋白的胞质/核定位。建立去卵巢（OVX）小鼠模型以验证体外研究结果。
结果
OVX 小鼠血清和股骨组织中 SNHG1 升高，miR-181c-5p 降低。SNHG1 通过上调 SFRP1 直接抑制 miR-181c-5p 以激活 Wnt3a/β-catenin 信号通路。此外，SNHG1 的敲低通过增加 miR-181c-5p 促进 BMSCs 的成骨分化。相反，SNHG1 的过表达促进 BMSCs 的破骨细胞分化并抑制 BM-EPCs 的血管生成，而过表达 miR-181c-5p 则逆转了这些作用。体内实验表明，SNHG1 沉默通过调节 miR-181c-5p 来刺激成骨生成和抑制破骨细胞生成，从而缓解骨质疏松症。重要的是，SP1 可在 BMSCs 中诱导 SNHG1。
结论
总之，SP1 诱导的 SNHG1 通过海绵吸附 miR-181c-5p 调节 SFRP1/Wnt/β-catenin 信号通路，从而抑制成骨细胞分化和血管生成，同时促进破骨细胞形成。此外，SNHG1 沉默可能为骨质疏松症提供一种潜在的治疗方法。
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