Zhenjie Qin1, Yuanye Li2, Yuanteng Li3, Guangyun Liu4. 1. Department of Stomatology, Zoucheng People's Hospital, Zoucheng, Shandong, People's Republic of China. 2. Office of Management of Hospital Infection, Jining No. 1 People's Hospital, Jining City, Shandong, People's Republic of China. 3. Department of Pharmacy, Zoucheng People's Hospital, Zoucheng, Shandong, People's Republic of China. 4. Department of Obstetrics and Gynecology, Zoucheng People's Hospital, Zoucheng, Shandong, People's Republic of China. Electronic address: zcskqkqzj@163.com.
Abstract
INTRODUCTION: It has been widely accepted that dental pulp stem cells (DPSCs), which are a class of self-renewal and differentiation potential of adult stem cells, play an important role in the repair procession of pulp's inflammation. We investigated whether tumor necrosis factor alpha (TNF-α) could induce the proliferation of DPSCs and clarified the potential mechanism of this proliferation. METHODS: Cell Counting Kit-8 assay (Dojindo Laboratories, Mashiki-machi, Kumamoto, Japan) and 5-ethynyl-2'-deoxyuridine-based proliferation assays were determined to investigate various concentrations or hours of TNF-α inducing a cell number change of DPSCs. Next, flow cytometry analysis was performed to investigate the main cell cycle phase process of DPSCs. Furthermore, the signaling pathway of TNF-α-induced proliferation of DPSCs was analyzed using Western blot analysis. Then, inhibitors were added to confirm the mechanism of this signaling pathway. RESULTS: TNF-α induced the proliferation of DPSCs in a dose- and time-dependent manner. Cyclin D1, which controlled the cell cycle process from the G1 to the S phase, was up-regulated by TNF-α in a time-dependent manner, whereas its overexpression alone increased DPSC proliferation. Furthermore, TNF-α was capable of inducing Akt/GSK-3β signaling pathway activation. Blockage of phosphoinositide 3-kinase/Akt by their kinase or genetic inhibitors could significantly reduce TNF-α-induced proliferation of DPSCs. CONCLUSIONS: This study confirmed that TNF-α induced the proliferation of DPSCs by regulating the Akt/GSK-3β/cyclin D1 signaling pathway and then provided a suitable number for the requirements of cell differentiation.
INTRODUCTION: It has been widely accepted that dental pulp stem cells (DPSCs), which are a class of self-renewal and differentiation potential of adult stem cells, play an important role in the repair procession of pulp's inflammation. We investigated whether tumor necrosis factor alpha (TNF-α) could induce the proliferation of DPSCs and clarified the potential mechanism of this proliferation. METHODS: Cell Counting Kit-8 assay (Dojindo Laboratories, Mashiki-machi, Kumamoto, Japan) and 5-ethynyl-2'-deoxyuridine-based proliferation assays were determined to investigate various concentrations or hours of TNF-α inducing a cell number change of DPSCs. Next, flow cytometry analysis was performed to investigate the main cell cycle phase process of DPSCs. Furthermore, the signaling pathway of TNF-α-induced proliferation of DPSCs was analyzed using Western blot analysis. Then, inhibitors were added to confirm the mechanism of this signaling pathway. RESULTS: TNF-α induced the proliferation of DPSCs in a dose- and time-dependent manner. Cyclin D1, which controlled the cell cycle process from the G1 to the S phase, was up-regulated by TNF-α in a time-dependent manner, whereas its overexpression alone increased DPSC proliferation. Furthermore, TNF-α was capable of inducing Akt/GSK-3β signaling pathway activation. Blockage of phosphoinositide 3-kinase/Akt by their kinase or genetic inhibitors could significantly reduce TNF-α-induced proliferation of DPSCs. CONCLUSIONS: This study confirmed that TNF-α induced the proliferation of DPSCs by regulating the Akt/GSK-3β/cyclin D1 signaling pathway and then provided a suitable number for the requirements of cell differentiation.