Exploration of the role of LncRNA PVT1 targeting miR-18b-5p in the differentiation of dental pulp mesenchymal stem cells into dentin

doi:10.19439/j.sjos.2026.01.006

Abstract

Abstract: PURPOSE: To analyze on the role of long-chain non coding ribonucleic acid (LncRNA) plasmacytoma variant translocation 1 (PVT1) in the differentiation of dental pulp mesenchymal stem cells (DPMSCs) into dentin by targeting microribonucleic acid 18b-5p(miR-18b-5p). METHODS: Human DPMSCs were isolated and cultured, then identified by staining with oil red O, Alizarin red, and Alisin blue, and detected cell surface marker molecules by flow cytometry. Inoculate cells onto a 24 well plate, 2×10⁴/cm² density, grouping, including PVT1 upregulation+miR-18b-5p downregulation group (transfected with pcDNA-PVT1 and antomiR-18b-5p), PVT1 downregulation+miR-18b-5p upregulation group (transfected with si-PVT1 and agomiR-18b-5p), empty 1 group (transfected with NC pcDNA), empty 2 group (transfected with ago-NC), PVT1 upregulation + empty 2 group (transfected with pcDNA-PVT1 and empty 2), empty1+miR-18b-5p downregulation group (transfected with empty1 and antomiR-18b-5p) and a blank group. After 7 days, Alizarin red staining was used to observe the calcium mineralization of cells, and alkaline phosphatase (ALP) activity was detected using a reagent kit. Real time fluorescence quantitative polymerase chain reaction(RT-qPCR) was used to detect the expressions of LncRNA PVT1, miR-18b-5p, dentin salivary phosphoprotein(DSPP), osteocalcin(OCN), and Runt related transcription factor 2 (RUNX2) genes in each group. Western blot(WB) was used to detect the expressions of DSPP, OCN, and RUNX2 proteins in each group. Dual luciferase reporter gene detection experiment was used to explore the targeting relationship between LncRNA PVT1 and miR-18b-5p. RESULTS: This study successfully isolated human DPMSCs and identified them by staining with oil red O, Alizarin red, Alishin blue and flow cytometry. The PVT1 upregulation+miR-18b-5p downregulation group had the strongest dentin differentiation activities (P<0.05), while that of PVT1 downregulation+miR-18b-5p upregulation group was the weakest(P<0.05). The LncRNA PVT1, DSPP, OCN and RUNX2 expressions were highest in the PVT1 upregulation+miR-18b-5p downregulation group(P<0.05), while miR-18b-5p expression was lowest (P<0.05). The PVT1 downregulation+miR-18b-5p upregulation group showed the opposite trend(P<0.05). There were binding sites between LncRNA PVT1 and miR-18b-5p, and LncRNA PVT1 could negatively feedback and target miR-18b-5p. CONCLUSIONS: Upregulation of LncRNA PVT1 can target the inhibition of miR-18b-5p and promote the differentiation of human DPMSCs into dentin, which may be related to the upregulation of DSPP, OCN and RUNX2 expressions.

Key words: Long-chain non coding ribonucleic acid, Plasmacytoma variant translocation 1, Microribonucleic acid 18b-5p, Dental pulp mesenchymal stem cells, Odontogenic differentiation

CLC Number:

R781.3

Zheng Yu, Zhai Tong, Cui Min, Liu Fei. Exploration of the role of LncRNA PVT1 targeting miR-18b-5p in the differentiation of dental pulp mesenchymal stem cells into dentin[J]. Shanghai Journal of Stomatology, 2026, 35(1): 35-41.

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