1. Academic Validation
  2. SOX2 suppresses osteoblast differentiation of MC3T3-E1 cells through activating the transcription of LGR4

SOX2 suppresses osteoblast differentiation of MC3T3-E1 cells through activating the transcription of LGR4

  • In Vitro Cell Dev Biol Anim. 2022 Dec 22. doi: 10.1007/s11626-022-00740-4.
Sunyu Chen 1 2 3 Zhanhao Xiao 4 Wenjin Jiang 4
Affiliations

Affiliations

  • 1 The Third Clinical Medical College, Fujian Medical University, No. 88, Jiaotong Road, Fuzhou, Fujian Province, 350004, People's Republic of China. hunson2022@163.com.
  • 2 Department of Orthopedics, Fuzhou Second Hospital, Cangshan District, No. 47, Shangteng Road, Fuzhou, Fujian Province, 350007, People's Republic of China. hunson2022@163.com.
  • 3 Fujian Provincial Clinical Medical Research Center for First Aid and Rehabilitation in Orthopaedic Trauma (2020Y2014), Fuzhou, People's Republic of China. hunson2022@163.com.
  • 4 Department of Orthopedics, Fuzhou Second Hospital, Cangshan District, No. 47, Shangteng Road, Fuzhou, Fujian Province, 350007, People's Republic of China.
Abstract

Osteogenic differentiation is a crucial process of new bone formation. This study aimed to explore the roles and mechanism of SRY-Box Transcription Factor 2 (SOX2) on proliferation and osteogenic differentiation of MC3T3-E1 cells. Bone Morphogenetic Protein 2 (BMP2) was used to induce the osteogenic differentiation of MC3T3-E1 cells. The expression of SOX2 was determined by quantitative Real-Time PCR (RT-PCR) at different time points after induction. The SOX2 overexpression plasmids were constructed and transfected into MC3T3-E1 cells. Osteogenic differentiation was evaluated by Alizarin Red S staining and Alkaline Phosphatase (ALP) assay. The expressions of osteogenic differentiation markers including runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) were detected by western blot assay. Luciferase reporter and CHIP assays were used to confirm that SOX2 regulated the transcriptional activation of leucine-rich repeat-containing G protein-coupled receptor 4 (LGR4). We found that SOX2 was down-regulated upon BMP2-induced osteogenic differentiation in MC3T3-E1 cells. Overexpression of SOX2 effectively inhibited osteogenic differentiation with decreased ALP activity, calcification, and osteogenic differentiation markers' expression including Runx2, OPN, and OCN. LGR4 was identified as a target of SOX2, and the inhibitory effect of SOX2 on osteogenic differentiation was reversed by knockdown of LGR4. The present study confirmed that SOX2 suppressed osteogenic differentiation of MC3T3-E1 cells through targeting LGR4, which possesses a therapeutic strategy for bone formation and generation.

Keywords

Bone formation; LGR4; MC3T3-E1; Osteogenic differentiation; SOX2.

Figures
Products