2025, Vol. 29 ›› Issue (36): 7776-7782

Long noncoding RNA TP53TG1 promotes odontogenic and osteogenic differentiation of stem cells from the apical papilla

Li Tingyue1, 2, 3, Guo Qian2, He Wenxi3, Wu Jiayuan1   

Abstract: BACKGROUND: Long noncoding RNA TP53TG1 (lncRNA TP53TG1) is involved in regulating the proliferation, migration, invasion, and apoptosis of various cancer cells, but there are few reports on its role in other cells.
OBJECTIVE: To investigate the effects and pathways of lncRNA TP53TG1 on the proliferation and differentiation of human stem cells from the apical papilla.
METHODS: Human stem cells from the apical papilla were isolated and cultured, and then transfected with lncRNA TP53TG1 overexpression lentivirus. RT-qPCR was used to detect the overexpression efficiency of lncRNA TP53TG1. Western blot assay was used to detect the relative expression levels of PI3K, AKT, ERK, P38, Smad3, and their phosphorylated proteins. Human stem cells from the apical papilla were divided into the empty lentiviral vector transfection group and the lncRNA TP53TG1 overexpression group. CCK-8 assay was used to measure the cell proliferation. Alkaline phosphatase activity was detected by alkaline phosphatase staining on day 5 of osteogenic induction. Formation of mineralized nodules was detected by alizarin red staining on day 21 of osteogenic induction. RT-qPCR was used to detect the mRNA expression levels of dentin sialophosphoprotein, Runt-related transcription factor 2, dentin matrix protein 1, and bone sialoprotein on days 3, 7, and 14 of osteogenic induction. Western blot assay was used to detect the protein expression levels of dentin sialophosphoprotein and Runt-related transcription factor 2 on days 3, 7, and 14 of osteogenic induction.
RESULTS AND CONCLUSION: (1) RT-qPCR results showed that the lentivirus was successfully integrated into the genome of stem cells from the apical papilla. Western blot assay results showed that overexpression of lncRNA TP53TG1 up-regulated the protein levels of p-PI3K and p-AKT without affecting the expression of phosphorylated proteins in other pathways. (2) Starting from day 3 of cell culture, overexpression of lncRNA TP53TG1 significantly promoted the proliferation of stem cells from the apical papilla. (3) In the process of inducing odontogenic differentiation of stem cells from the apical papilla, overexpression of lncRNA TP53TG1 promoted the expression of odontogenic and osteogenic differentiation-related genes and proteins, significantly increased alkaline phosphatase activity and mineralized nodule formation. (4) The results show that lncRNA TP53TG1 may promote the odontogenic and osteogenic differentiation of stem cells from the apical papilla by activating the PI3K/AKT signaling pathway.

Key words: stem cell from the apical papilla, long noncoding RNA, lncRNA TP53TG1, virus transfection, dentinogenesis, PI3K/AKT signaling pathway, engineered stem cell