Overexpression of ID1 reverses the repression of human dental pulp stem cells differentiation induced by TWIST1 silencing
Abstract
Multiple studies showed that the cessation of TWIST1 expression is the prerequisite for osteoblasts’ maturation. However, recent reports revealed that TWIST1 works differently in the dental pulp stem cells (DPSCs), where a high level of TWIST1 expression promoted DPSCs’ differentiation. Objective was to investigate impact of TWIST1 and ID1 on the differentiation process in the human DPSCs.
Methods: TWIST1 and ID1 expression in the DSPCs was modulated by lentivirus transduction. Genes expression was assessed with qRT-PCR. The proteins level was evaluated by Western blot. The DPSCs differentiation was assessed with the proliferation, alkaline phosphatase (ALP) activity, and calcium concentration assays.
Results: TWIST1 silencing caused the suppression of the expression of ID1 and both the early and late markers of odontoblasts’ differentiation detected at the transcripts and proteins level. The forced overexpression of ID1 increased the expression of the late markers of odontoblasts differentiation but diminished the expression of the early markers. DPCSs with the silenced TWIST1 and subsequent ID1 overexpression displayed an increase in the expression of the late markers of odontoblasts differentiation. Cells with silenced TWIST1 and overexpressing ID1 had increased activity of ALP, higher calcium concentration and decreased proliferation rate. The high level of ID1 expression might be a critical factor stimulating DPSCs to differentiation and it might compensate the repressed differentiation of DPSCs caused by TWIST1 silencing. Conclusion: The intriguing interaction between the expression of both the TWIST1 and ID1 might be a critical factor driving the process of the human odontoblasts’ differentiation.
References
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