DNMT3A对骨关节炎软骨细胞SOX9基因DNA甲基化修饰的影响  

Effect of DNMT3A on DNA methylation modification of SOX9 gene in osteoarthritis chondrocytes

作　　者：霍少川 杨俊兴 刘付懿斐 郭富明 程文昌 温刘莹 钟嘉漫 蒋子云 Huo Shaochuan;Yang Junxing;Liufu Yifei;Guo Fuming;Cheng Wenchang;Wen Liuying;Zhong Jiaman;Jiang Ziyun(Department of Orthopedics&Traumatology,Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen Guangdong,518000;Guangzhou University of Chinese Medicine,Guangzhou Guangdong,510006,China)

机构地区：[1]广州中医药大学深圳医院(福田)骨伤科,广东深圳518000 [2]广州中医药大学,广东广州510006

出　　处：《生物骨科材料与临床研究》2023年第4期29-34,共6页Orthopaedic Biomechanics Materials and Clinical Study

基　　金：国家自然科学基金青年项目(82004395);中国博士后科学基金(2018M633088)及出站博士后科研资助;深圳市福田区卫生公益性科研项目(FTWS2020059)

摘　　要：目的探讨DNMT3A对SOX9基因DNA甲基化修饰的影响。方法培养正常软骨细胞和OA软骨细胞,显微镜下观察软骨细胞形态,甲苯胺蓝染色、Ⅱ型胶原染色鉴定软骨细胞,qRT-PCR、Western Blot检测DNMT3A、SOX9 mRNA和蛋白表达水平,亚硫酸氢盐测序检测SOX9基因DNA甲基化状态,慢病毒沉默DNMT3A观察SOX9基因DNA甲基化修饰的变化。结果正常软骨细胞形态规则,体积较大,数量较多,OA软骨细胞形态不规则,体积较小,数量较少。与正常软骨细胞比较,OA软骨细胞中DNMT3A mRNA和蛋白表达量增高(P<0.05),SOX9 mRNA和蛋白表达量明显降低(P<0.05),SOX9基因DNA甲基化百分比增高。在OA软骨细胞中沉默DNMT3A后,SOX9基因DNA甲基化百分比明显降低。结论DNMT3A调节SOX9基因DNA甲基化修饰参与OA软骨细胞病变。Objective To investigate the effect of DNMT3A on DNA methylation modification of the SOX9 gene.Methods Normal chondrocytes and OA chondrocytes were cultured.The morphology of chondrocytes was observed by microscope.Toluidine blue staining and typeⅡcollagen staining were used to identify chondrocytes.DNMT3A,SOX9 mRNA and protein expression levels were detected by qRT-PCR and Western Blot.The methylation status of SOX9 promoter was detected by bisulfite sequencing.Dnmt3A was silenced by lentivirus to observe the changes of DNA methylation modification of SOX9 gene.Results Normal chondrocytes had regular morphology,large volume and large number,while OA chondrocytes had irregular morphology,small volume and small number.Compared with normal chondrocytes,the expression of DNMT3A mRNA and protein in OA chondrocytes increased(P<0.05),the expression of SOX9 mRNA and protein decreased significantly(P<0.05),and the percentage of DNA methylation of SOX9 gene increased.After DNMT3A silencing in OA chondrocytes,the percentage of DNA methylation of SOX9 gene was significantly decreased.Conclusion DNA methylation modification of SOX9 gene regulated by DNMT3A is involved in OA chondrocyte lesions.

关 键 词：骨关节炎 软骨细胞 DNMT3A SOX9 DNA甲基化 

分 类 号：R318.08[医药卫生—生物医学工程]