海南医学院学报2024,Vol.30Issue(11) :811-816,823.DOI:10.13210/j.cnki.jhmu.20240307.001

甲基转移酶METTL3介导m6A甲基化调控犬JBMMSCs成骨分化

Methyltransferase METTL3 regulates osteogenic differentiation of canine JBMMSCs by mediating m6A methylation

陈翌 蒋丽花 黄旋平
海南医学院学报2024,Vol.30Issue(11) :811-816,823.DOI:10.13210/j.cnki.jhmu.20240307.001

甲基转移酶METTL3介导m6A甲基化调控犬JBMMSCs成骨分化

Methyltransferase METTL3 regulates osteogenic differentiation of canine JBMMSCs by mediating m6A methylation

陈翌 1蒋丽花 1黄旋平1
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作者信息

  • 1. 广西医科大学口腔医学院/附属口腔医院口腔颌面外科,广西 南宁 530021;广西口腔颌面修复与重建研究自治区级重点实验室,广西 南宁 530021;广西颅颌面畸形临床医学研究中心,广西 南宁 530021
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摘要

目的:探究甲基转移酶METTL3 对颌骨骨髓间充质干细胞(Jaw bone marrow mesenchymal stem cells,JBMMSCs)增殖、迁移及成骨分化的影响.方法:体外培养JBMMSCs,慢病毒转染构建沉默METTL3基因细胞模型,转染后siMETTL3组JBMMSCs mRNA水平(P<0.001)及蛋白水平(P<0.01)下降.检测两组细胞的增殖、迁移、碱性磷酸酶活性、钙沉积以及成骨相关因子骨钙素(osteocalcin,OCN)mRNA及蛋白水平.结果:与沉默对照(siNC)组相比,siMETTL3组培养24、48、72、96 h的增殖活性降低(P<0.001),24 h后细胞迁移能力下降(P<0.05).成骨诱导7 d后siMETTL3组碱性磷酸酶活性下降,21 d后矿化沉积较siNC组少.qRT-PCR及Western Blot显示,OCN的mRNA(P<0.001)和蛋白(P<0.05)表达水平降低.结论:沉默METTL3可抑制JBMMSCs的增殖、迁移及成骨分化.

Abstract

Objective:To explore the effect of methyltransferase METTL3 on the proliferation,migration and osteogenic dif-ferentiation of jaw bone marrow mesenchymal stem cells(JBMMSCs).Methods:JBMMSCs were cultured in vitro and transfect-ed with lentivirus to construct a cell model of silencing METTL3 gene.After transfection,the mRNA level(P<0.001)and pro-tein level(P<0.01)of JBMMSCs in the siMETTL3 group decreased.The proliferation,migration,alkaline phosphatase activi-ty,calcium deposition,and osteogenesis-related factors osteocalcin(OCN)mRNA and protein level of the two groups of JBMMSCs were detected.Results:Compared with the siNC group,the proliferation activity of the siMETTL3 group decreased after 24,48,72,and 96 h of culture(all P<0.001),and the cell migration ability decreased after 24 h(P<0.05).The activity of alkaline phosphatase in the siMETTL3 group decreased after 7 days of osteogenic induction,and the mineralization deposition af-ter 21 days was less than that in the siNC group.qRT-PCR and Western blot showed that the mRNA level(P<0.001)and protein expression level(P<0.05)of OCN reduced.Conclusion:Silencing METTL3 can inhibit the proliferation,migration and osteo-genic differentiation of JBMMSCs.

关键词

METTL3/颌骨骨髓间充质干细胞/成骨分化/m6A甲基化

Key words

METTL3/Jaw bone marrow mesenchymal stem cells/Osteogenic differentiation/m6A methylation

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基金项目

国家自然科学基金(82360187)

广西壮族自治区医疗卫生适宜技术开发与推广应用项目(S2021085)

广西科技基地和人才专项(2021AC18031)

南宁市青秀区科技计划(2021004)

出版年

2024
海南医学院学报
海南医学院

海南医学院学报

CSTPCD北大核心
影响因子:1.068
ISSN:1007-1237
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