LncRNA PART1/miR-185-5p/RUNX3 反馈环调节骨髓间充质干细胞的成骨分化。

LncRNA PART1/miR-185-5p/RUNX3 feedback loop modulates osteogenic differentiation of bone marrow mesenchymal stem cells.

机构信息

Department of Orthopedics and Traumatology, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu, China.

Department of Spinal Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu, China.

出版信息

Autoimmunity. 2021 Nov;54(7):422-429. doi: 10.1080/08916934.2021.1966771. Epub 2021 Aug 25.

DOI:10.1080/08916934.2021.1966771

PMID:34431433

Abstract

BACKGROUND

Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is essential for bone formation, and its dysfunction is reported to be associated with osteoporosis (OP). Recent researches have determined that lncRNA PART1 participates in the pathogenesis of multiple diseases. However, its role in modulating osteogenic differentiation of hBMSCs is unclear.

METHODS

PART1, miR-185-5p, and RUNX3 levels were assessed RT-qPCR. The protein levels of OCN, OSN, and COL1A1 were measured by western blotting. The osteoblastic phenotype was evaluated ALP activity and ARS staining. The relationship between miR-185-5p and PART1 or RUNX3 was validated by luciferase reporter, RIP assays.

RESULTS

PART1 and RUNX3 expression were enhanced during hBMSC osteogenic differentiation. PART1 deletion decreased OCN, OSN, and COL1A1 levels and weakened ALP activity, but promoted the apoptosis of hBMSCs. Moreover, PART1 served as a ceRNA to influence the RUNX3 level targeting miR-185-5p. In addition, RUNX3 was verified to activate the transcription of PART1 in hBMSCs. Finally, rescue assays indicated that suppression of miR-185-5p or addition of RUNX3 partially abolished the effects of PART1 knockdown on the levels of OCN, OSX, and COL1A1 levels, ALP activity, and apoptosis.

CONCLUSION

Our study elaborated that PART1/miR-185-5p/RUNX3 feedback contributed to osteogenic differentiation and inhibited the hBMSCs apoptosis, suggesting that PART1 might be a novel target for OP treatment.

摘要

背景

人骨髓间充质干细胞（hBMSCs）的成骨分化对于骨形成至关重要，其功能障碍与骨质疏松症（OP）有关。最近的研究确定长链非编码 RNA PART1 参与多种疾病的发病机制。然而，其在调节 hBMSCs 成骨分化中的作用尚不清楚。

方法

采用 RT-qPCR 检测 PART1、miR-185-5p 和 RUNX3 的水平。采用 Western blot 法检测 OCN、OSN 和 COL1A1 的蛋白水平。通过碱性磷酸酶（ALP）活性和 ARS 染色评估成骨表型。通过荧光素酶报告、RIP 测定验证 miR-185-5p 与 PART1 或 RUNX3 之间的关系。

结果

PART1 和 RUNX3 的表达在 hBMSC 成骨分化过程中增强。PART1 缺失降低了 OCN、OSN 和 COL1A1 水平，并减弱了 ALP 活性，但促进了 hBMSCs 的凋亡。此外，PART1 作为 ceRNA 通过靶向 miR-185-5p 影响 RUNX3 水平。此外，RUNX3 被证实可在 hBMSCs 中激活 PART1 的转录。最后，挽救实验表明，抑制 miR-185-5p 或添加 RUNX3 部分消除了 PART1 敲低对 OCN、OSX 和 COL1A1 水平、ALP 活性和凋亡的影响。