LSD1 knockdown confers protection against osteoclast formation by reducing histone 3 lysine 9 monomethylation and dimethylation in ITGB3 promoter

IF 2.3 4区 生物学 Q4 CELL BIOLOGY
Dongping Yu , Zhen Li , Jie Cao , Guowen Wei , Feng Shen
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Abstract

ITGB3, an osteoclast marker, is involved in osteoclast formation. Nevertheless, its related mechanism remains poorly characterized. Herein, this study examines the mechanisms affecting osteoclast formation with the involvement of ITGB3. Osteoclast formation was induced with macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-kappa B ligand (RANKL), followed by measurement of the mRNA and protein expression of ITGB3 and LSD1. After gain- and loss-of-function assays, cell viability and the expression of osteoclast marker genes (NFATc1, ACP5, and CTSK) were assessed, and osteoclast formation was evaluated with TRAP staining. ChIP assays were used to examine histone 3 lysine 9 (H3K9) monomethylation (H3K9me1) and H3K9 dimethylation (H3K9me2) modifications and LSD1 protein enrichment in the ITGB3 promoter. During osteoclast formation, ITGB3 and LSD1 were gradually augmented. Knockdown of LSD1 or ITGB3 curbed cell viability, the expression of osteoclast marker genes, and osteoclast formation. Moreover, overexpression of ITGB3 nullified the suppressive impact of LSD1 knockdown on osteoclast formation. Mechanistically, LSD1 promoted ITGB3 expression by reducing H3K9 levels in the ITGB3 promoter. LSD1 enhanced ITGB3 expression by decreasing H3K9me1 and H3K9me2 levels in ITGB3 promoter to boost osteoclast formation.

LSD1敲低通过减少ITGB3启动子中的组蛋白3赖氨酸9单甲基化和二甲基化来提供对破骨细胞形成的保护。
ITGB3是一种破骨细胞标志物,参与破骨细胞的形成。然而,其相关机制的特点仍然很差。在此,本研究探讨了ITGB3参与破骨细胞形成的机制。巨噬细胞集落刺激因子(M-CSF)和核因子κB受体激活剂配体(RANKL)诱导破骨细胞形成,然后测量ITGB3和LSD1的mRNA和蛋白表达。在功能获得和丧失测定后,评估细胞活力和破骨细胞标记基因(NFATc1、ACP5和CTSK)的表达,并用TRAP染色评估破骨细胞的形成。ChIP测定用于检测组蛋白3赖氨酸9(H3K9)单甲基化(H3K9me1)和H3K9二甲基化(H3K9me2)修饰以及ITGB3启动子中的LSD1蛋白富集。在破骨细胞形成过程中,ITGB3和LSD1逐渐增强。LSD1或ITGB3的敲除抑制了细胞活力、破骨细胞标记基因的表达和破骨细胞的形成。此外,ITGB3的过表达抵消了LSD1敲低对破骨细胞形成的抑制作用。从机制上讲,LSD1通过降低ITGB3启动子中的H3K9水平来促进ITGB3的表达。LSD1通过降低ITGB3启动子中的H3K9me1和H3K9me2水平来增强ITGB3的表达,以促进破骨细胞的形成。
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来源期刊
Acta histochemica
Acta histochemica 生物-细胞生物学
CiteScore
4.60
自引率
4.00%
发文量
107
审稿时长
23 days
期刊介绍: Acta histochemica, a journal of structural biochemistry of cells and tissues, publishes original research articles, short communications, reviews, letters to the editor, meeting reports and abstracts of meetings. The aim of the journal is to provide a forum for the cytochemical and histochemical research community in the life sciences, including cell biology, biotechnology, neurobiology, immunobiology, pathology, pharmacology, botany, zoology and environmental and toxicological research. The journal focuses on new developments in cytochemistry and histochemistry and their applications. Manuscripts reporting on studies of living cells and tissues are particularly welcome. Understanding the complexity of cells and tissues, i.e. their biocomplexity and biodiversity, is a major goal of the journal and reports on this topic are especially encouraged. Original research articles, short communications and reviews that report on new developments in cytochemistry and histochemistry are welcomed, especially when molecular biology is combined with the use of advanced microscopical techniques including image analysis and cytometry. Letters to the editor should comment or interpret previously published articles in the journal to trigger scientific discussions. Meeting reports are considered to be very important publications in the journal because they are excellent opportunities to present state-of-the-art overviews of fields in research where the developments are fast and hard to follow. Authors of meeting reports should consult the editors before writing a report. The editorial policy of the editors and the editorial board is rapid publication. Once a manuscript is received by one of the editors, an editorial decision about acceptance, revision or rejection will be taken within a month. It is the aim of the publishers to have a manuscript published within three months after the manuscript has been accepted
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