Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Department of Bone Metabolism, School of Stomatology Shandong University, Jinan, 250012, China.
Department of Developmental Biology of Hard Tissue, Graduate School of Dental Medicine, Hokkaido University, Sapporo, 060-8586, Japan.
Biochem Biophys Res Commun. 2019 Jun 11;513(4):1019-1026. doi: 10.1016/j.bbrc.2019.04.112. Epub 2019 Apr 19.
Dexamethasone (DEX), a widely used glucocorticoid with strong anti-inflammatory and immunosuppressive activities, has been reported to induce apoptosis in osteoblasts, but the underlying mechanisms are still not comprehensively investigated. FOXO1 plays an important role in the regulation of cell proliferation and apoptosis. Our study aims to explore the role of FOXO1 in DEX-induced apoptosis of osteoblastic MC3T3-E1 cells through bioinformatics and experiments. We first employed bioinformatics to identify DEX-related genes and revealed their functions by GO enrichment analysis including FOXO1 associated biological processes. Expression level of FOXO1 was validated by GEO data. Then, experiments were performed to verify the hypothesis. CCK8 was used to detect cell viability and apoptosis was detected by flow cytometry. SiRNA was used to silence FOXO1 and western-blot was employed to detect protein expression. Results demonstrated DEX-related genes involved in cell proliferation, apoptosis and angiogenesis and FOXO1 was a regulator of apoptosis. DEX could up-regulate FOXO1 expression, inhibit cell viability, promote apoptosis. SiRNA-FOXO1 could attenuate DEX-induced apoptosis in MC3T3-E1. These findings suggested DEX could affect some vital biological processes of MC3T3-E1 and FOXO1 played an essential role in DEX-induced apoptosis in MC3T3-E1.
地塞米松（DEX）是一种广泛应用的糖皮质激素，具有很强的抗炎和免疫抑制作用，据报道可诱导成骨细胞凋亡，但其中的机制尚不完全清楚。FOXO1 在细胞增殖和凋亡的调节中起着重要作用。我们的研究旨在通过生物信息学和实验探讨 FOXO1 在 DEX 诱导的成骨细胞 MC3T3-E1 细胞凋亡中的作用。我们首先利用生物信息学方法识别 DEX 相关基因，并通过 GO 富集分析揭示其功能，包括与 FOXO1 相关的生物学过程。通过 GEO 数据验证 FOXO1 的表达水平。然后进行实验验证假设。CCK8 用于检测细胞活力，流式细胞术检测细胞凋亡。使用 siRNA 沉默 FOXO1，Western blot 检测蛋白表达。结果表明，DEX 相关基因参与细胞增殖、凋亡和血管生成，FOXO1 是凋亡的调节因子。DEX 可上调 FOXO1 表达，抑制细胞活力，促进凋亡。siRNA-FOXO1 可减弱 DEX 诱导的 MC3T3-E1 细胞凋亡。这些发现表明 DEX 可能影响 MC3T3-E1 的一些重要生物学过程，FOXO1 在 DEX 诱导的 MC3T3-E1 细胞凋亡中起着重要作用。
