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miRNA-211 表达在后骨折骨细胞凋亡中的作用涉及转化生长因子-β/磷脂酰肌醇 3-激酶信号通路。

The function of microRNA-211 expression in post-fracture bone cell apoptosis involving the transforming growth factor-β/ phosphoinositide 3-kinase signaling pathway.

机构信息

Department of Orthopedics, Dongying People's Hospital, Dongying, Shandong province, P. R. China.

Department of Orthopedics, Dongying District People's Hospital, Dongying, Shandong province, P. R. China.

出版信息

J Int Med Res. 2020 Jul;48(7):300060520926353. doi: 10.1177/0300060520926353.

DOI:10.1177/0300060520926353
PMID:32720595
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7388126/
Abstract

BACKGROUND

The underlying mechanism of micro (mi)RNA-211 in bone cell apoptosis after fracture remains unclear. This study aimed to determine the effect and function of miRNA-211 in bone cell apoptosis in fracture patients.

METHODS

Serum samples were collected from patients with fractures and healthy controls. Serum miR-211 expression was detected by quantitative PCR. MC3T3-E1 cells were transfected with a transforming growth factor (TGF)-β inhibitor and phosphoinositide 3-kinase (PI3K) inhibitor. The viability of MC3T3-E1 cells was detected by the MTT assay, and apoptosis was detected by flow cytometry. Caspase-3/9 activity and the protein expression of TGF-β, PI3K, and p-Akt were detected by western blot and immunoprecipitation.

RESULTS

In the fracture group, miRNA-211 expression was significantly up-regulated compared with controls. We used miRNA-211 mimics to up-regulate miRNA-211 expression, and observed inhibited cell viability and induced apoptosis and lactate dehydrogenase (LDH) activity. miRNA-211 up-regulation also suppressed the expression of TGF-β, PI3K, and p-Akt proteins. Conversely, miRNA-211 down-regulation increased cell viability and reduced apoptosis and LDH activity, as well as inducing the expression of TGF-β, PI3K, and p-Akt. Inhibiting TGF-β decreased the effect of anti-miRNA-211 on osteocyte apoptosis.

CONCLUSION

Our data indicate that miRNA-211 functions via the TGF-β/PI3K/Akt signaling pathway in patients with fractures.

摘要

背景

微小 RNA-211（miRNA-211）在骨折后骨细胞凋亡中的作用机制尚不清楚。本研究旨在探讨 miRNA-211 在骨折患者骨细胞凋亡中的作用和功能。

方法

收集骨折患者和健康对照者的血清样本。采用实时定量 PCR 检测血清 miR-211 表达。用转化生长因子（TGF）-β抑制剂和磷脂酰肌醇 3-激酶（PI3K）抑制剂转染 MC3T3-E1 细胞。MTT 法检测 MC3T3-E1 细胞活力，流式细胞术检测细胞凋亡。Western blot 和免疫沉淀法检测 caspase-3/9 活性及 TGF-β、PI3K、p-Akt 蛋白表达。

结果

与对照组相比，骨折组中 miRNA-211 表达明显上调。我们用 miRNA-211 模拟物上调 miRNA-211 表达，观察到细胞活力受到抑制，细胞凋亡和乳酸脱氢酶（LDH）活性增加。miRNA-211 上调还抑制了 TGF-β、PI3K 和 p-Akt 蛋白的表达。相反，miRNA-211 下调增加了细胞活力，减少了细胞凋亡和 LDH 活性，同时诱导了 TGF-β、PI3K 和 p-Akt 的表达。抑制 TGF-β 降低了抗 miRNA-211 对成骨细胞凋亡的作用。

结论

我们的数据表明，miRNA-211 通过 TGF-β/PI3K/Akt 信号通路在骨折患者中发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/4c8626668b3c/10.1177_0300060520926353-fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/e5938572ba46/10.1177_0300060520926353-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/434069bbed19/10.1177_0300060520926353-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/66660c14410c/10.1177_0300060520926353-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/40178a54a1d2/10.1177_0300060520926353-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/65df9275fd08/10.1177_0300060520926353-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/4c8626668b3c/10.1177_0300060520926353-fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/e5938572ba46/10.1177_0300060520926353-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/434069bbed19/10.1177_0300060520926353-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/66660c14410c/10.1177_0300060520926353-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/40178a54a1d2/10.1177_0300060520926353-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/65df9275fd08/10.1177_0300060520926353-fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6f1/7388126/4c8626668b3c/10.1177_0300060520926353-fig6.jpg

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