转录因子SP3在人牙髓干细胞成骨向分化中的作用研究

doi:10.13701/j.cnki.kqyxyj.2026.03.009

摘要/Abstract

摘要： 目的:本研究旨在探讨转录因子特异性蛋白3(specificity protein 3,SP3)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)成骨向分化的调控作用。方法:体外分离并培养hDPSCs,通过体外多向分化诱导,对其多向分化能力进行鉴定;通过定量实时逆转录聚合酶链反应(quantitative real-time reverse transcription polymerase chain reaction,qRT-PCR)检测SP3在hDPSCs成骨向分化不同时期的动态表达;利用小干扰RNA(small interfering RNA,siRNA)技术下调hDPSCs中SP3的表达;在成骨诱导培养条件下,通过qRT-PCR和Western blot检测Runt相关转录因子2(runt-related transcription factor 2,RUNX2)和骨桥蛋白(osteopontin,OPN)等成骨相关标志物的表达情况。同时,通过碱性磷酸酶(alkaline phosphatase,ALP)染色和茜素红S(alizarin red S,ARS)染色,对hDPSCs的成骨向分化潜能进行评估。结果:SP3在hDPSCs成骨向分化过程中呈时间依赖性上调;成骨向诱导后,抑制SP3表达的hDPSCs中,RUNX2和OPN的基因及蛋白表达水平均下调,ALP和ARS染色强度也均减弱。结论:抑制表达SP3可降低hDPSCs的成骨向分化潜能。

关键词: SP3, 人牙髓干细胞, 成骨向分化

Abstract: Objective: To investigate the role of the transcription factor SP3 in the osteogenic differentiation of human dental pulp stem cells (hDPSCs) in vitro. Methods: hDPSCs were isolated and cultured in vitro, and their multipotent differentiation potential was identified through induction of multilineage differentiation. The expression pattern of SP3 was analyzed using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). SP3 expression was knocked down by small interfering RNA (siRNA) transfection, followed by osteogenic induction. The osteogenic differentiation potential was analyzed by alkaline phosphatase (ALP) and alizarin red S (ARS) staining, as well as by qRT-PCR and Western blot analysis for osteogenic markers including runt-related transcription factor 2 (RUNX2) and osteopontin (OPN). Results: SP3 expression was found to increase in a time-dependent fashion during the osteogenic induction process of hDPSCs. Knockdown of SP3 in hDPSCs significantly reduced ALP and ARS staining and downregulated the expression of RUNX2 and OPN. Conclusion: Suppression of SP3 expression inhibits the osteogenic differentiation of hDPSCs.

Key words: specificity protein 3, human dental pulp stem cells, osteogenic differentiation

肖爽, 李炅, 刘馨杨, 宋帅, 张莉, 张杰. 转录因子SP3在人牙髓干细胞成骨向分化中的作用研究[J]. 口腔医学研究, 2026, 42(3): 225-229.

XIAO Shuang, LI Jiong, LIU Xinyang, SONG Shuai, ZHANG Li, ZHANG Jie. Role of Transcription Factor SP3 in Osteogenic Differentiation of Human Dental Pulp Stem Cells[J]. Journal of Oral Science Research, 2026, 42(3): 225-229.

参考文献

[1] Zhao RS, Yang RJ, Cooper PR, et al. Bone grafts and substitutes in dentistry: a review of current trends and developments[J]. Molecules, 2021, 26(10):3007.
[2] Zhang J, Zhang WH, Yue WJ, et al. Research progress of bone grafting: A comprehensive review[J]. Int J Nanomedicine, 2025, 20:4729-4757.
[3] Sui B, Wu D, Xiang L, et al. Dental pulp stem cells: from discovery to clinical application[J]. J Endod, 2020, 46(9S): S46-S55.
[4] 夏亮,江文欣,邹多宏.牙髓干细胞多项分化潜能的研究及应用进展[J].医学综述, 2022, 28(11):2086-2091.
[5] Papineni S, Chintharlapalli S, Abdelrahim M, et al. Tolfenamic acid inhibits esophageal cancer through repression of specificity proteins and c-Met[J]. Carcinogenesis, 2009, 30(7):1193-1201.
[6] Wang X, Sun K, Xu Z, et al. Roles of SP/KLF transcription factors in odontoblast differentiation: From development to diseases[J]. Oral Dis, 2024, 30(6):3745-3760.
[7] Zhou W, Fang J, Jia Q, et al. Transcription factor specificity protein (SP) family in renal physiology and diseases[J]. PeerJ, 2025, 13: e18820.
[8] Göllner H, Dani C, Phillips B, et al. Impaired ossification in mice lacking the transcription factor Sp3[J]. Mech Dev, 2001, 106(1-2):77-83.
[9] He YL, Chen SJ, Xia DS, et al. Neural crest-derived mesenchymal stem cells: Fates and perspectives[J]. World J Stem Cells, 2025, 17(9):107689.
[10] 于乐蓉,李祥伟,艾虹.牙髓干细胞干性维持的研究进展[J].国际口腔医学杂志,2023,50(4):463-471.
[11] Huang L, Chen X, Yang XX, et al. Elucidating epigenetic mechanisms governing odontogenic differentiation in dental pulp stem cells: an in-depth exploration[J]. Front Cell Dev Biol, 2024, 12:1394582.
[12] Zhong L, Sun Y, Wang C, et al. SP1 regulates BMSC osteogenic differentiation through the miR-133a-3p/MAPK3 axis: SP1 regulates osteogenic differentiation of BMSCs[J]. J Orthop Surg Res, 2024, 19(1):396.
[13] Lang J, Morya VK, Kwak MK, et al. Molecular crosstalk in SP7-mediated osteogenesis: Regulatory mechanisms and therapeutic potential[J]. Osteoporos Sarcopenia, 2025, 11(2):31-37.
[14] McCoy AM, Lakhdari O, Shome S, et al. Sp3 is essential for normal lung morphogenesis and cell cycle progression during mouse embryonic development[J]. Development, 2023, 150(5): dev200839.
[15] Harris S, Anwar I, Baksh SS, et al. Skeletal muscle differentiation induces wide-ranging nucleosome repositioning in muscle gene promoters[J]. Sci Rep, 2024, 14(1):9396.
[16] Zhu Q, Liao K. Differential expression of the adipocyte amino acid transporter is transactivated by SP1 and SP3 during the 3T3-L1 preadipocyte differentiation process[J]. Biochem Biophys Res Commun, 2000, 271(1):100-106.
[17] Baksh SS, Pratt RE, Gomez J, et al. A novel Cbx1, PurB, and Sp3 complex mediates long-term silencing of tissue- and lineage-specific genes[J]. J Biol Chem, 2022, 298(6):102053.
[18] Kim HJ, Shin HR, Yoon H, et al. Peptidylarginine deiminase 2 plays a key role in osteogenesis by enhancing RUNX2 stability through citrullination[J]. Cell Death Dis, 2023, 14(8):576.
[19] Khanna-Jain R, Mannerström B, Vuorinen A, et al. Osteogenic differentiation of human dental pulp stem cells on β-tricalcium phosphate/poly (l-lactic acid/caprolactone) three-dimensional scaffolds[J]. J Tissue Eng, 2012, 3(1):2041731412467998.
[20] Cianferotti L. Osteomalacia is not a single disease[J]. Int J Mol Sci, 2022, 23(23):14896.
[21] Zheng BY, Geng YQ, Li Y, et al. Specificity protein 1/3 regulate T-cell acute lymphoblastic leukemia cell proliferation and apoptosis through β-catenin by acting as targets of miR-495-3p[J]. Ann Hematol, 2024, 103(8):2945-2960.
[22] Lu H, Yuan P, Ma X, et al. Angiotensin-converting enzyme inhibitor promotes angiogenesis through Sp1/Sp3-mediated inhibition of notch signaling in male mice[J]. Nat Commun, 2023, 14(1):731.