长链非编码 RNA HAGLR 通过负调控 miRNA-19a-3p 加速股骨颈骨折愈合。

LncRNA HAGLR accelerates femoral neck fracture healing through negatively regulating miRNA-19a-3p.

机构信息

Department of Orthopedics, Ningbo Medical Center Lihuili Eastern Hospital, Ningbo, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Apr;24(8):4080-4087. doi: 10.26355/eurrev_202004_20984.

Abstract

OBJECTIVE

This study aims to uncover the function of long non-coding RNA (lncRNA) HAGLR in the healing process of femoral neck fracture and the underlying mechanism.

PATIENTS AND METHODS

Expression levels of HAGLR, microRNA-19a-3p (miRNA-19a-3p) and TGFBR2 in fractured femoral neck tissues and adjacent normal tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Regulatory effects of HAGLR on viability, apoptosis, migration, and protein levels of BALP and Osteocalcin in MC3T3-E1 cells were determined. Dual-Luciferase reporter gene assay was conducted to assess the binding in HAGLR/miRNA-19a-3p/TGFBR2. In addition, relative levels of TGFBR2, p-smad2, p-smad3, and RUNX2 in MSCs influenced by HAGLR were detected.

RESULTS

HAGLR was downregulated in fractured femoral neck tissues. Knockdown of HAGLR reduced viability and migration, enhanced apoptotic rate, as well as downregulated BALP and Osteocalcin in MC3T3-E1 cells. HAGLR served as miRNA-19a-3p sponge, and miRNA-19a-3p directly targeted 3'-untranslated region (3'-UTR) of TGFBR2. Knockdown of HAGLR downregulated expressions of TGFBR2, p-smad2, p-smad3, and RUNX2 in MC3T3-E1 cells, indicating the inhibited TGF-β pathway.

CONCLUSIONS

LncRNA HAGLR/miRNA-19a-3p/TGFBR2 regulatory loop accelerates the healing process of femoral neck fracture by inhibiting the TGF-β pathway.

摘要

目的

本研究旨在揭示长链非编码 RNA(lncRNA)HAGLR 在股骨颈骨折愈合过程中的作用及其潜在机制。

方法

采用实时定量聚合酶链反应(qRT-PCR)检测骨折股骨颈组织及相邻正常组织中 HAGLR、微小 RNA-19a-3p(miRNA-19a-3p)和 TGFBR2 的表达水平。检测 HAGLR 对 MC3T3-E1 细胞活力、凋亡、迁移及碱性磷酸酶和骨钙素蛋白水平的影响。采用双荧光素酶报告基因实验评估 HAGLR/miRNA-19a-3p/TGFBR2 的结合情况。此外,还检测了 HAGLR 对 MSCs 中 TGFBR2、p-smad2、p-smad3 和 RUNX2 相对水平的影响。

结果

HAGLR 在骨折股骨颈组织中表达下调。敲低 HAGLR 降低了 MC3T3-E1 细胞的活力和迁移能力,促进了细胞凋亡,并下调了碱性磷酸酶和骨钙素蛋白的表达。HAGLR 作为 miRNA-19a-3p 的海绵,miRNA-19a-3p 直接靶向 TGFBR2 的 3'-非翻译区(3'-UTR)。敲低 HAGLR 下调了 MC3T3-E1 细胞中 TGFBR2、p-smad2、p-smad3 和 RUNX2 的表达,表明 TGF-β 通路受到抑制。

结论

lncRNA HAGLR/miRNA-19a-3p/TGFBR2 调控环通过抑制 TGF-β 通路加速股骨颈骨折的愈合过程。

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