Mechanism of XCT-790 reducing activity and osteogenesis of osteoblasts

doi:10.3969/j.issn.2095-4344.0600

Abstract

Abstract:

BACKGROUND: Wnt signaling pathway plays an important role in formation, differentiation and mature of osteocytes. As negative regulators of Wnt signaling pathway, DKK1 and Sost play negative roles in regulating bone mass and osteoblast differentiation. XCT-790, an inhibitor of estrogen receptor-related receptor α, can regulate the effect of estrogen signaling pathway on osteoclast function.
OBJECTIVE: To observe the effects of XCT-790 on the proliferation of MG63 cells transfected with DKK 1 and Sost overexpression adenovirus vector and the expression of low-density lipoprotein receptor-related protein (LRP) 5, bone morphologic protein 2, osteopontin, osteoprotegerin, and connective tissue growth factor (CTGF) protein.
METHODS: MG63 cells were divided into eight groups: blank control, overexpressed DKK1 (ad-DKK1), overexpressed Sost (ad-Sost), overexpressed DKK1+Sost (ad-DKK1+Sost), XCT-790-intervened blank adenovirus, XCT-790-intervened ad-Sost, XCT-790-intervened ad-DKK1-Sost groups. The cell viability was detected by MTT assay. The alkaline phosphatase activity was detected by alkaline phosphatase kit. Ca2+ concentration was detected by flow cytometer. Expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF was detected by western blot assay.
RESULTS AND CONCLUSION: Compared with the blank control group, ad-DKK1, ad-Sost, ad-DKK1+Sost and XCT-790-intervened blank adenovirus groups could significantly decrease the cell activity, alkaline phosphatase activity, increase calcium concentration and down-regulate the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF (P < 0.05). In the XCT-790-intervented ad-DKK1, ad-Sost, ad-DKK1+Sost groups, the cell activity, alkaline phosphatase activity, and the expression of LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin and CTGF were significantly reduced, and the Ca2+ concentration was significantly increased (P < 0.05). In summary, XCT-790 can directly regulate Wnt signaling pathway, and can cooperate with DKK1 and Sost to inhibit Wnt signaling pathway, thereby decreasing the activity of osteoblasts, LRP5, bone morphologic protein 2, osteopontin, osteoprotegerin, and CTGF protein expression, and further decreasing differentiation, proliferation and osteogenesis of osteoblasts.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Estrogen Receptor alpha, Adenoviridae, Transfection, Tissue Engineering

CLC Number:

R446

Liu Shaojin1, Wan Lei2, Qiao Rongqin2, Huang Hongxing2, Wang Jili1, Li Zhaozheng1 . Mechanism of XCT-790 reducing activity and osteogenesis of osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(3): 329-334.

Figures/Tables 3

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