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

作　　者：陈翌 蒋丽花 黄旋平[1,2,3] CHEN Yi;JIANG Lihua;HUANG Xuanping(Department of Oral and Maxillofacial Surgery,College&Hospital of Stomatology,Guangxi Medical University,Nanning 530021,China;Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Nanning 530021,China;Guangxi Clinical Research Center for Craniofacial Deformity,Guangxi Medical University,Nanning 530021,China)

机构地区：[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 [2]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 [3]广西颅颌面畸形临床医学研究中心,广西南宁530021

出　　处：《海南医学院学报》2024年第11期811-816,823,共7页Journal of Hainan Medical University

基　　金：国家自然科学基金资助项目(82360187);广西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)。

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。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 

分 类 号：R782.4[医药卫生—口腔医学]