赖氨酸乙酰基转移酶2A通过经典Wnt通路影响牙周膜干细胞成骨分化

doi:10.7518/hxkq.2018.01.008

华西口腔医学杂志 ›› 2018, Vol. 36 ›› Issue (1): 39-45.doi: 10.7518/hxkq.2018.01.008

赖氨酸乙酰基转移酶2A通过经典Wnt通路影响牙周膜干细胞成骨分化

郭武城¹(), 程洁莉¹, 杨征毅¹, 张忆¹, 何恩亮¹, 钱钧¹, 宋晶晶¹, 孙晋², 袁林¹()

1.广州医科大学附属第一医院口腔科,广州 510120
2.南方医科大学附属深圳市妇幼保健院口腔病防治中心,深圳 518048

收稿日期:2017-06-25 修回日期:2017-10-12 出版日期:2018-02-07 发布日期:2018-02-01
作者简介:
郭武城,硕士研究生,E-mail:574143236@qq.com
基金资助:
[基金项目] 广州市科信局科技惠民专项基金（2014Y2-00058）;广州市属高校科研计划科技类一般项目（2012C096）

K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway

Wucheng Guo¹(), Jieli Cheng¹, Zhengyi Yang¹, Yi Zhang¹, Enliang He¹, Jun Qian¹, Jingjing Song¹, Jin Sun², Lin Yuan¹()

1. Dept. of Stomatology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China
2. Stomatological Disease Center, Shenzhen Maternal and Child Health Care Hospital, Southern Medical University, Shenzhen 518048, China

Received:2017-06-25 Revised:2017-10-12 Online:2018-02-07 Published:2018-02-01
Supported by:
Supported by: Science and Technology Huimin Special Fund of Guangzhou Municipal Science and Information Bureau (2014Y2-00058);General Project of Scientific Research Program of University of Guangzhou (2012C096).

摘要/Abstract

摘要：

目的探讨赖氨酸乙酰基转移酶2A（KAT2A）调控牙周膜干细胞（PDLSCs）成骨分化的机制。方法分离培养来自健康志愿者的PDLSCs（H-PDLSCs）和牙周炎患者的PDLSCs（P-PDLSCs）,比较传代细胞中KAT2A基因的表达水平。采用KAT2A基因干扰H-PDLSCs,检测KAT2A基因干扰对H-PDLSCs成骨分化的影响;同时检测KAT2A干扰后对经典Wnt通路及其配体的影响,确定KAT2A基因与经典Wnt通路的上下游关系。结果与H-PDLSCs相比,P-PDLSCs中KAT2A基因的表达下降,差异有统计学意义（P<0.05）。KAT2A基因被干扰后,PDLSCs成骨能力下降（P<0.05）,同时经典Wnt通路被激活,拮抗剂Dickkopf-1（DKK-1）表达下调;加入DKK-1后,被干扰的PDLSCs成骨分化功能恢复,而KAT2A的表达不受影响,仍维持较低水平。结论牙周炎可导致PDLSCs中KAT2A基因表达下降,激活经典Wnt通路,抑制细胞的成骨分化。

关键词: 牙周膜干细胞, 牙周炎, 乙酰基转移酶, 成骨分化, 经典Wnt通路

Abstract:

Objective This study aims to investigate the mechanism of K (lysine) acetyltransferase 2A (KAT2A) regulation and control on the osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Methods The expression levels of KAT2A in PDLSCs were compared from each generation of the normal (H-PDLSCs) and periodontitis tissues (P-PDLSCs). The influences of KAT2A gene interference on the osteogenic differentiation of PDLSCs were also detected. In addition, the influences of the KAT2A gene interference to the canonical Wnt pathway and ligands were detected. The upstream and down-stream relationships between KAT2A and canonical Wnt pathway were also determined. Results The decreased expression of KAT2A in PDLSCs from the inflammatory tissue in each generation was compared with that in PDLSCs from the healthy tissue, and the difference was statistically significant (P<0.05). When the KAT2A gene was disrupted, the osteogenesis ability of PDLSC was declined, and the difference was statistically significant (P<0.05). The canonical Wnt pathway was activated, and the antagonist Dickkopf-1 (DKK-1) was reduced. After the DKK-1 addition, the osteogenic differentiation of the disturbed PDLSCs was recovered, and KAT2A was unaffected. Conclusion The KAT2A expression in PDLSCs was decreased because of perio-dontitis. The classical Wnt pathway was activated to inhibit the osteogenic differentiation of the cells.

Key words: periodontal ligament stem cells, perio-dontitis, acetyltransferase, osteogenic differentiation, classical Wnt pathway

中图分类号:

R781

郭武城, 程洁莉, 杨征毅, 张忆, 何恩亮, 钱钧, 宋晶晶, 孙晋, 袁林. 赖氨酸乙酰基转移酶2A通过经典Wnt通路影响牙周膜干细胞成骨分化[J]. 华西口腔医学杂志, 2018, 36(1): 39-45.

Wucheng Guo, Jieli Cheng, Zhengyi Yang, Yi Zhang, Enliang He, Jun Qian, Jingjing Song, Jin Sun, Lin Yuan. K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway[J]. West China Journal of Stomatology, 2018, 36(1): 39-45.

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参考文献 17

[1]	Pittenge MF, Macka AM, Bec SC, et al.Multilineage potential of adult human mesenchymal stem cells[J]. Science, 1999
	284(5411):143-147.
[2]	Uccelli A, Pistoia V, Moretta L.Mesenchymal stem cells: a new strategy for immunosuppression[J]. Trends Immunol, 2007, 28(5):219-226.
[3]	Liu Y, Liu W, Hu C, et al.MiR-17 modulates osteogenic differentiation through a coherent feed-forward loop in me-senchymal stem cells isolated from periodontal ligaments of patients with periodontitis[J]. Stem Cells, 2011, 29(11): 1804-1816.
[4]	Liu N, Shi S, Deng M, et al.High levels of β-catenin sig-naling reduce osteogenic differentiation of stem cells in inflammatory microenvironments through inhibition of the noncanonical Wnt pathway[J]. J Bone Miner Res, 2011, 26(9):2082-2095.
[5]	孔祥伟, 叶瑞东, 刘文佳, 等. 炎症微环境中经典Wnt信号通路对牙周膜干细胞成骨分化的调控作用[J]. 口腔生物医学, 2015, 6(3):129-136.
	Kong XW, Ye RD, Liu WJ, et al.Wnt/β-catenin signaling pathway mediates the impaired osteogenic differentiation of periodontal ligament stem cells in inflammatory microen-vironment[J]. Oral Biomed, 2015, 6(3):129-136.
[6]	崔晓霞, 杨琨, 伍燕, 等. 糖基化终末产物介导的牙周膜干细胞成骨分化过程中Wnt经典信号通路相关作用机制的研究[J]. 牙体牙髓牙周病学杂志, 2013, 23(9):564-568, 680.
	Cui XX, Yang K, Wu Y, et al.Effects of advanced glyca-tion end products on the osteogenic influence of wnt classic pathway in human periodontal ligament stem cells[J]. Chin J Conserv Dent, 2013, 23(9):564-568, 680.
[7]	谢金芳, 张泽兵. 自噬的表观遗传修饰在肿瘤发生发展中的作用[J]. 现代肿瘤医学, 2016, 24(13):2156-2159.
	Xie JF, Zhang ZB. The role of epigenetic modifications on autophagy in tumor[J].J Mod Oncol, 2016,24(13):2156-2159
[8]	Li C, Li B, Dong Z, et al.Lipopolysaccharide differentially affects the osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells through Toll-like receptor 4 mediated nuclear factor κB pathway[J]. Stem Cell Res Ther, 2014, 5(3):67.
[9]	Timmermann S, Lehrmann H, Polesskaya A, et al.Histone acetylation and disease[J]. Cell Mol Life Sci, 2001, 58(5):
	728-736.
[10]	Grant PA, Duggan L, Côté J, et al.Yeast Gcn5 functions in two multisubunit complexes to acetylate nucleosomal his-tones: characterization of an Ada complex and the SAGA (Spt/Ada) complex[J]. Genes Dev, 1997, 11(13):1640-1650.
[11]	王露霏, 白丁, 韩向龙. Wnt信号通路拮抗剂Dkk1调节骨代谢的最新进展[J]. 中国组织工程研究, 2013, 17(2):337-341-
12	Wang LF, Bai D, Han XL.Progress in dickkopf-1-mediated bone metabolism[J]. J Clin Rehabil Tissue Eng Res, 2013, 17(2):337-341.
[12]	Pihlstrom BL, Michalowicz BS, Johnson NW.Periodontal diseases[J]. Lancet, 2005, 366(9499):1809-1820.
[13]	Gomez RS, Dutra WO, Moreira PR.Epigenetics and perio-dontal disease: future perspectives[J]. Inflamm Res, 2009, 58(10):625-629.
[14]	Ye L, Fan Z, Yu B, et al.Histone demethylases KDM4B and KDM6B promotes osteogenic differentiation of human MSCs[J]. Cell Stem Cell, 2012, 11(1):50-61.
[15]	Liu W, Konermann A, Guo T, et al.Canonical Wnt signaling differently modulates osteogenic differentiation of mesen-chymal stem cells derived from bone marrow and from periodontal ligament under inflammatory conditions[J]. Biochim Biophys Acta, 2014, 1840(3):1125-1134.
[16]	Semënov MV, Tamai K, Brott BK, et al.Head inducer Dick-kopf-1 is a ligand for Wnt coreceptor LRP6[J]. Curr Biol, 2001, 11(12):951-961.
[17]	Mao B, Wu W, Davidson G, et al.Kremen proteins are Dick-kopf receptors that regulate Wnt/beta-catenin signalling[J]. Nature, 2002, 417(6889):664-667.

基因名称	引物序列（5’—3’）
GAPDH	F：CTGCAAGAACAGCATTGCAT
	R：GACCACCTGGTCCTCAGTGT
KAT2A	F：CCAAATCAAGTCTCACCCCAGT
	R：GGAAGCGGATGACCTCGTAG
WNT1	F：CGGGCAACAACCAAAGTCG
	R：CGTTCACAATACCCCACCAT
WNT2	F：ACAGCAGGCCGTGTGTGTGCAA
	R：AGGCAGTCCTGACAGCGCAC
WNT2B	F：GACCGGGACCACACCGTCTTTGG
	R：GGTGGAGGGTGGAGGAAGGTG
WNT3A	F：CTCGGATACTTCTTACTCCTCTGC
	R：CCTGGATGCCAATCTTGATG
WNT8A	F：GATGCCAGAGCCCTGATGAA
	R：GCCTGGTCATACTTGGCCTT
WNT10A	F：TTCTTCCTACTGCTGCTGGC
	R：TTAGGCACACTGTGTTGGCA
WNT10B	F：CAACACCGTGTGCTTGACG
	R：CAGCCAGCATGGAGAAGGAA
DKK-1	F：GGGAATTACTGCAAAAATGGAATA
	R：ATGACCGGAGACAAACCAGAAC
骨硬化蛋白	F：AGCTGGAGAACAACAAGACCA
	R：CATCGGTCACGTAGCGGG
Sfrp1	F：GATGCAGGAGGCTCAGGTGAT
	R：GCTGGCAACAGGTCAGAACG

干细胞标记物	H-PDLSCs	P-PDLSCs
Ctr-PE（PE通道对照组）	0.11	0.13
Ctr-FITC（FITC通道对照组）	0.08	0.05
STRO-1	31.24	37.61
CD146	74.92	77.17
CD105	80.52	78.34
CD90	98.35	99.08
CD34	0.31	0.26
CD14	0.21	0.14

细胞传代数	H-PDLSCs	P-PDLSCs	P值
P0	1.00±0.15	0.62±0.13	<0.01
P2	1.00±0.18	0.62±0.05	<0.01
P4	1.00±0.11	0.68±0.06	<0.05
P6	1.00±0.09	0.71±0.07	<0.05

赖氨酸乙酰基转移酶2A通过经典Wnt通路影响牙周膜干细胞成骨分化

K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway

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