A preliminary study on the impact of Notch1 on RANKL/RANK system in osteoclast via RAW264.7 cells
10.3969/j.issn.1674-8115.2018.12.008
- Author:
Jun WANG
1
Author Information
1. Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases with Integrated Chinese-Western Medicine, Shanghai Institute of Traumatology and Orthopaedics, Shanghai Jiao Tong University School of Medicine
- Publication Type:Journal Article
- Keywords:
Bone resorption;
Notch pathway;
Osteoclast;
RANKL/RANK system
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2018;38(12):1440-1446
- CountryChina
- Language:Chinese
-
Abstract:
Objective • To investigate the crosstalk between receptor activator of nuclear factor-κB ligand (RANKL) / receptor activator of nuclear factor-κB (RANK) system and Notch signaling pathways. Methods • Mouse osteoclast precursor cell line RAW264.7 was cultured and stimulated by RANKL (35 ng/L). Real-time PCR and Western blotting analysis were used to determine the expressions of Notch signaling molecules in RANKL-stimulated RAW264.7 cells. Small interfering RNA (siRNA) transfection was used to inhibit the expression of Notch1, Dll1 and Dll3 in Notch signaling pathways, then the mRNA and protein expressions of Rank were detected using real-time PCR and Western blotting analysis, respectively. Results • The expressions of Notch1, Jagged1 and Jagged2 were down-regulated while the expression of Dll4 was up-regulated after RANKL stimulation. The stimulation also inhibited the expression of Hes1 and Hey1. After siRNA transfection, the mRNA expressions of Notch1, Dll1 and Dll3 were suppressed by 71% (P=0.000), 53% (P=0.015) and 70% (P=0.000), respectively. The mRNA and protein expressions of Rank were up-regulated after Notch1 inhibition (P=0.003, P=0.000). Conclusion • The data infers that the impact of Notch1 on RANKL/RANK system might play a key role in bone resorption conducted by osteoclast.
