长链非编码 RNA HULC 通过下调 miR-195 促进骨髓间充质干细胞的增殖和成骨分化。
Long non-coding RNA HULC promotes proliferation and osteogenic differentiation of bone mesenchymal stem cells via down-regulation of miR-195.
机构信息
Department of Orthopedics, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 May;22(10):2954-2965. doi: 10.26355/eurrev_201805_15050.
OBJECTIVE
LncRNAs HULC has been reported to be important regulators in the development of various human diseases. However, the role of HULC in bone mesenchymal stem cells (BMSCs) remains unclear. The present study aimed to explore the regulatory effect of HULC on proliferation and osteogenic differentiation of BMSCs and the underlying mechanism.
MATERIALS AND METHODS
The expression of HULC and miR-195 in BMSCs were altered by transfection and measured by qRT-PCR. Cell viability was measured by the CCK-8 assay. Osteogenic differentiation of BMSCs was determined by evaluation of osteogenic markers (Ocn, ALP, Runx2, and Col-1) expression levels using Western blot and qRT-PCR. Furthermore, Western blot was performed to assess the expression of proliferation-related factors, Wnt/β-catenin and p38MAPK pathway-related factors.
RESULTS
HULC overexpression significantly increased cell viability, down-regulated p21 expression but up-regulated CyclinD1 expression, and promoted the levels of osteogenic markers. However, the complete opposite effect was observed in HULC knockdown. Notably, miR-195 expression was negatively regulated by HULC and miR-195 exerted a reversed effect of HULC on BMSCs. Moreover, miR-195 mediated the regulatory effect of HULC on BMSCs proliferation and osteogenic differentiation, as miR-195 mimic abolished the effect of HULC overexpression on BMSCs. We also found that HULC overexpression enhanced the activation of Wnt/β-catenin and p38MAPK pathway through down-regulating miR-195.
CONCLUSIONS
We revealed that HULC promoted proliferation and osteogenic differentiation of BMSCs. The potential mechanism might be involved in its negative regulation on miR-195 and enhanced activation of Wnt/β-catenin and p38MAPK pathway.
目的
已有研究报道长链非编码 RNA（lncRNA）HULC 是多种人类疾病发生发展的重要调控因子。然而，HULC 在骨髓间充质干细胞（BMSCs）中的作用尚不清楚。本研究旨在探讨 HULC 对 BMSCs 增殖和成骨分化的调控作用及其潜在机制。
材料和方法
通过转染改变 BMSCs 中 HULC 和 miR-195 的表达，并采用 qRT-PCR 进行检测。通过 CCK-8 法检测细胞活力。通过 Western blot 和 qRT-PCR 评估成骨标志物（Ocn、ALP、Runx2 和 Col-1）表达水平来确定 BMSCs 的成骨分化情况。此外，通过 Western blot 检测增殖相关因子、Wnt/β-catenin 和 p38MAPK 通路相关因子的表达。
结果
HULC 过表达显著增加细胞活力，下调 p21 表达但上调 CyclinD1 表达，并促进成骨标志物水平。然而，在 HULC 敲低时则观察到完全相反的效果。值得注意的是，HULC 负调控 miR-195 的表达，miR-195 对 BMSCs 发挥与 HULC 相反的作用。此外，miR-195 介导了 HULC 对 BMSCs 增殖和成骨分化的调节作用，因为 miR-195 模拟物消除了 HULC 过表达对 BMSCs 的影响。我们还发现，HULC 过表达通过下调 miR-195 增强了 Wnt/β-catenin 和 p38MAPK 通路的激活。
结论
我们揭示了 HULC 促进 BMSCs 的增殖和成骨分化。其潜在机制可能涉及 HULC 对 miR-195 的负调控以及 Wnt/β-catenin 和 p38MAPK 通路的增强激活。
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