沉默Versican V0/V1对小鼠牙乳头细胞生物学行为的影响

doi:10.13591/j.cnki.kqyx.2024.05.006

Abstract

Abstract:

Objective To investigate the effect of Versican V0/V1 on biological behaviors of mouse dental papilla cells (mDPCs). Methods mDPCs were isolated from C57BL/6J mice at embryonic day 16.5 (E16.5). A small interfering RNA (siRNA) constructed specifically for Versican V0/V1 was transfected into mDPCs.The silencing efficiency was verified by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) and immunofluorescence staining. The proliferation rate of mDPCs was determined using EdU assay; the migration ability of mDPCs was evaluated by scratch test and transwell assay. Alkaline phosphatase (ALP) staining and alizarin red staining were used to assess the mineralization capability of mDPCs. The molecules related to odontogenic differentiation and mineralization at mRNA levels were measured by qRT-PCR. Results After siRNA transfection,the mDPCs of si-Versican V0/V1 group showed weaker proliferation and migration abilities compared with si-NC group (P<0.01). An enhanced ALP staining intensity,mineralized nodule formation and up-regulations of the molecules related to odontogenic differentiation and mineralization at mRNA levels (P<0.05) were observed in the mDPCs of si-Versican V0/V1 group. Conclusion Versican V0/V1 silencing inhibits the proliferation and migration of mDPCs,but enhances the abilities of odontogenic differentiation and mineralization.

Key words: Versican, mouse dental papilla cell, cell proliferation, cell migration, odontogenic differentiation

CLC Number:

WU Jiayan, HUANG Haiyan, SONG Chenyu, JIANG Beizhan. Effects of Versican V0/V1 gene silencing on biological behaviors of mouse dental papilla cells[J]. Stomatology, 2024, 44(5): 349-355.

Figures/Tables 7

Tab.1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

References 21

[1]	Tavakoli S, Evans A, Oommen OP, et al. Unveiling extracellular matrix assembly: Insights and approaches through bioorthogonal chemistry[J]. Mater Today Bio, 2023, 22: 100768.
[2]	Tziafas D, Kodonas K. Differentiation potential of dental papilla, dental pulp, and apical papilla progenitor cells[J]. J Endod, 2010, 36(5):781-789. doi: 10.1016/j.joen.2010.02.006 pmid: 20416419
[3]	Foulcer SJ, Day AJ, Apte SS. Isolation and purification of versican and analysis of versican proteolysis[J]. Methods Mol Biol, 2022, 2303: 559-578. doi: 10.1007/978-1-0716-1398-6_43 pmid: 34626407
[4]	Islam S, Watanabe H. Versican: A dynamic regulator of the extracellular matrix[J]. J Histochem Cytochem, 2020, 68(11):763-775. doi: 10.1369/0022155420953922 pmid: 33131383
[5]	Xu FF, Cai WP, Chen WT, et al. Expression of different isoforms of versican during the development of mouse mandibular first molars[J]. J Histochem Cytochem, 2019, 67(7):471-480. doi: 10.1369/0022155419846875 pmid: 31034318
[6]	Hernández D, Miquel-Serra L, Docampo MJ, et al. Role of versican V0/V1 and CD44 in the regulation of human melanoma cell behavior[J]. Int J Mol Med, 2011, 27(2):269-275. doi: 10.3892/ijmm.2010.577 pmid: 21136024
[7]	Arslan F, Bosserhoff AK, Nickl-Jockschat T, et al. The role of versican isoforms V0/V1 in glioma migration mediated by transforming growth factor-beta2[J]. Br J Cancer, 2007, 96(10):1560-1568.
[8]	Sutherland TE, Dyer DP, Allen JE. The extracellular matrix and the immune system: A mutually dependent relationship[J]. Science, 2023, 379(6633):eabp8964.
[9]	Santinha D, Vilaça A, Estronca L, et al. Remodeling of the cardiac extracellular matrix proteome during chronological and pathological aging[J]. Mol Cell Proteomics, 2024, 23(1):100706.
[10]	Listik E, Azevedo Marques Gaschler J, Matias M, et al. Proteoglycans and dental biology: The first review[J]. Carbohydr Polym, 2019, 225: 115199.
[11]	Tang FY, Brune JE, Chang MY, et al. Defining the versican interactome in lung health and disease[J]. Am J Physiol Cell Physiol, 2022, 323(2):C249-C276.
[12]	Li HM, Ghorbani S, Zhang RY, et al. Prominent elevation of extracellular matrix molecules in intracerebral hemorrhage[J]. Front Mol Neurosci, 2023, 16: 1251432.
[13]	Yeung TL, Leung CS, Wong KK, et al. TGF-β modulates ovarian cancer invasion by upregulating CAF-derived versican in the tumor microenvironment[J]. Cancer Res, 2013, 73(16):5016-5028.
[14]	Ruggeri A, Orsini G, Mazzoni A, et al. Immunohistochemical and biochemical assay of versican in human sound predentine/dentine matrix[J]. Eur J Histochem, 2009, 53(3):e15.
[15]	Jiang BZ, Yokohama-Tamaki T, Wang ZL, et al. Expression, localisation and synthesis of versican by the enamel organ of developing mouse molar tooth germ: An in vivo and in vitro study[J]. Arch Oral Biol, 2010, 55(12):995-1006.
[16]	Bigoni S, Neri M, Scotton C, et al. Homozygous recessive versican missense variation is associated with early teeth loss in a Pakistani family[J]. Front Genet, 2018, 9: 723. doi: 10.3389/fgene.2018.00723 pmid: 30740127
[17]	Sheng W, Wang GZ, Wang Y, et al. The roles of versican V1 and V2 isoforms in cell proliferation and apoptosis[J]. Mol Biol Cell, 2005, 16(3):1330-1340. doi: 10.1091/mbc.e04-04-0295 pmid: 15635104
[18]	蒋备战, 柴田俊一, 王佐林. Versican在ICR胎鼠下颌第一磨牙牙胚发育过程中的表达[J]. 实用口腔医学杂志, 2009, 25(5):626-629.
[19]	Guo SL, Lim D, Dong ZH, et al. Dentin sialophosphoprotein: A regulatory protein for dental pulp stem cell identity and fate[J]. Stem Cells Dev, 2014, 23(23):2883-2894. doi: 10.1089/scd.2014.0066 pmid: 25027178
[20]	Vimalraj S, Arumugam B, Miranda PJ, et al. Runx2: Structure, function, and phosphorylation in osteoblast differentiation[J]. Int J Biol Macromol, 2015, 78: 202-208. doi: 10.1016/j.ijbiomac.2015.04.008 pmid: 25881954
[21]	Vimalraj S. Alkaline phosphatase: Structure, expression and its function in bone mineralization[J]. Gene, 2020, 754: 144855.

基因	引物序列(5'-3')
Versican V0	F:ATGAAGACGGAGAGGAGGACTGTG
Versican V0	R:CACTGTGGTAACGAGCTGCTTCC
Versican V1	F:GCAGATTTGATGCCTACTGC R:ACGGAGTAGTTGTTACATCCG
Alp	F:CACAGACTCCCTGCTTTTGCT R:TGCCTCAGAAGAGCTGAAAACTT
Dspp	F:GTGGGATCATCAGCCAGTCAG R:TGCCTTTGTTGGGACCTTCA
Runx2	F:ATGCATTTAAGATATGGTTGCC R:TGGAGTTGGGAAACACTTGA
Gapdh	F:GACATCAAGAAGGTGGTGAAGC R:GAAGGTGGAAGAGTGGGAGTT

Effects of Versican V0/V1 gene silencing on biological behaviors of mouse dental papilla cells

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 21

Related Articles 8

Recommended Articles

Metrics

Comments

[1]	. Progress of research on exosomes from dental tissue-derived stem cells [J]. , 2020, 40(12): 1147-1151.
[2]	. Effect of dealing with pure titanium implant surface on adhesion and proliferation of human gingival fibroblast [J]. , 2019, 39(12): 1135-1139.
[3]	. The protective effects of hyaluronic acid against nicotine in human periodontal ligament fibroblasts [J]. , 2018, 38(4): 314-319.
[4]	. Effect of the growth differentiation factor 15 on rat bone marrowderived mesenchymal stem cell proliferation and gene expression [J]. , 2017, 37(9): 778-784.
[5]	FAN Zhipeng. Controlling the odontogenic differentiation of mesenchymal stem cells: the key to dental tissue regeneration [J]. , 2017, 37(2): 97-101.
[6]	. MicroRNA27b inhibits cell proliferation of oral squamous cell carcinoma through targeting FZD7 [J]. , 2016, 36(3): 200-205.
[7]	. The effect of 1064nm Nd:YAG laser irradiation on the migration of oral squamous cell carcinoma CAL-27 cells [J]. , 2016, 36(2): 109-111.
[8]	. Study on effect of Chloride channel blocker NPPB on the proliferation and differentiation of chicken mandibular mesenchymal cells [J]. , 2015, 35(6): 438-441.