Journal of Oral Science Research ›› 2021, Vol. 37 ›› Issue (5): 448-452.DOI: 10.13701/j.cnki.kqyxyj.2021.05.014

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DDIT3 Regulates Chondrogenic Differentiation of ATDC5 Cells Under TNF-α Stimulation

YANG Chang, XU Xiaoxiao, DONG Xiaofei, DONG Wei, WANG Jiawei*   

  1. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China
  • Received:2021-01-08 Published:2021-05-17

Abstract: Objective: To investigate the function of DDIT3 in the regulation of chondrogenic differentiation of ATDC5 cells under TNF-α stimulation. Methods: Wild-type ATDC5 cells were induced to chondrogenic differentiation for 7 days under the stimulation of various concentrations of TNF-α (0, 1, 10, 20 ng/mL), and then lentivirus-transfected ATDC5 cells were treated with chondrogenic induction medium contained 10ng/mL TNF-α for 7 days. qRT-PCR was used to detect mRNA changes of inflammation-related genes COX2, IL-6, NOS2, and MMP-13. Western blot was utilized to evaluate the protein expression levels of DDIT3 and chondrogenic related markers Sox9, Col10a1 p-AKT, and AKT. Alcian blue staining was conducted to detect extracellular matrix. Results: The expression of DDIT3 and inflammation-related factors COX2, IL-6, NOS2, and MMP-13 were increased in ATDC5 cells under chondrogenic induction in various concentrations of TNF-α after 7 days, while the chondrogenic differentiation markers Sox9 and Col10a1 were decreased. Knockdown of DDIT3 partly reversed the decreased expression levels of Sox9 and Col10a1 and extracellular matrix formation in ATDC5 cells under TNF-α stimulation, while promoted the expression level of p-AKT. Conclusion: Under TNF-α induced inflammatory microenvironment, DDIT3 could regulate chondrogenic differentiation of ATDC5 cells

Key words: DDIT3, TNF-α, ATDC5 cells, chondrogenic differentiation