长链非编码 RNA HOTTIP 通过与 TAF15 相互作用稳定 DLX2 促进骨髓间充质干细胞成骨分化并诱导血管生成。
LncRNA HOTTIP facilitates osteogenic differentiation in bone marrow mesenchymal stem cells and induces angiogenesis via interacting with TAF15 to stabilize DLX2.
机构信息
Department of Pathology and Pathophysiology, School of Basic Medicine, Guizhou Medical University, Guiyang, 550002, Guizhou Province, PR China; Department of Prosthodontics, The Affiliated Stomatology Hospital of Guizhou Medical University, Guiyang, 550002, Guizhou Province, PR China; Guizhou Medical University School of Stomatology, Guiyang, 550002, Guizhou province, PR China.
Department of Prosthodontics, The Affiliated Stomatology Hospital of Guizhou Medical University, Guiyang, 550002, Guizhou Province, PR China; Guizhou Medical University School of Stomatology, Guiyang, 550002, Guizhou province, PR China.
出版信息
Exp Cell Res. 2022 Aug 15;417(2):113226. doi: 10.1016/j.yexcr.2022.113226. Epub 2022 May 26.
AIM
The molecular mechanism of differentiation in bone marrow mesenchymal stem cells (BMSCs) preserves to be further elucidated. LncRNA HOTTIP has been proven to accelerate osteogenic differentiation, but the regulation mechanism is still unclear.
METHODS
The human BMSCs (hBMSCs) were isolated and identified by the antigen CD29, CD34, CD44, CD45, and CD90 through flow cytometry. The osteogenic state was determined by the ALP Detection Kit and Alizarin red staining. The tube formation was observed under a microscope. HOTTIP expression level, DLX2 and TAF15, Wnt/β-catenin pathway, and transcriptional markers in osteogenesis and angiogenesis were examined with Western blot and RT-qPCR, respectively. The combination of TAF15 with lncRNA HOTTIP and DLX2 was detected by RNA immunoprecipitation (RIP) and RNA pulldown assays.
RESULTS
The outcomes revealed that HOTTIP was noticeably up-regulated accompanied by the osteogenic transcriptional factor in the process of osteoblast differentiation and angiogenesis. Besides, HOTTIP enhanced alkaline phosphatase (ALP) activity, accelerated osteogenic differentiation and angiogenesis along with up-regulation of osteogenic and angiogenic-related gene expression, by interaction with TAF15 to stabilize DLX2.
CONCLUSION
Taken together, our outcomes reveal that lncRNA HOTTIP accelerated osteogenic differentiation and angiogenesis by interaction with TAF15 to stabilize DLX2.
目的
骨髓间充质干细胞（BMSCs）分化的分子机制仍需进一步阐明。长链非编码 RNA HOTTIP 已被证明能加速成骨分化，但调控机制尚不清楚。
方法
通过流式细胞术检测人骨髓间充质干细胞（hBMSCs）表面抗原 CD29、CD34、CD44、CD45 和 CD90 进行分离和鉴定。通过碱性磷酸酶（ALP）检测试剂盒和茜素红染色来确定成骨状态。在显微镜下观察管形成情况。用 Western blot 和 RT-qPCR 分别检测 HOTTIP 表达水平、成骨和血管生成中的 DLX2 和 TAF15、Wnt/β-catenin 通路以及转录标志物。用 RNA 免疫沉淀（RIP）和 RNA 下拉实验检测 TAF15 与 lncRNA HOTTIP 和 DLX2 的结合情况。
结果
研究结果表明，在成骨细胞分化和血管生成过程中，HOTTIP 明显上调，并伴有成骨转录因子。此外，HOTTIP 通过与 TAF15 相互作用稳定 DLX2，增强碱性磷酸酶（ALP）活性，加速成骨分化和血管生成，并上调成骨和血管生成相关基因的表达。
结论
综上所述，我们的研究结果表明，lncRNA HOTTIP 通过与 TAF15 相互作用稳定 DLX2，从而加速成骨分化和血管生成。
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