doi:10.3969/j.issn.1672-5972.2023.04.007文章编号:swgk2022-03-00080生物骨科材料与临床研究ORTHOPAEDICBIOMECHANICSMATERIALSANDCLINICALSTUDY2023年08月第20卷第4期DNMT3A对骨关节炎软骨细胞SOX9基因DNA甲基化修饰的影响*霍少川1杨俊兴1刘付懿斐1郭富明2程文昌2温刘莹2钟嘉漫2蒋子云1*[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、WesternBlot检测DNMT3A、SOX9mRNA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DNMT3AmRNA和蛋白表达量增高（P＜0.05），SOX9mRNA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。[关键词]骨关节炎；软骨细胞；DNMT3A；SOX9；DNA甲基化[中图分类号]R318.08[文献标识码]AEffectofDNMT3AonDNAmethylationmodificationofSOX9geneinosteoarthritischondrocytesHuoShaochuan1,YangJunxing1,LiufuYifei1,GuoFuming2,ChengWenchang2,WenLiuying2,ZhongJiaman2,JiangZiyun1.1DepartmentofOrthopedics&Traumatology,ShenzhenHospitalofGuangzhouUniversityofChineseMedicine(Futian),ShenzhenGuangdong,518000;2GuangzhouUniversityofChineseMedicine,GuangzhouGuangdong,510006,China[Abstract]ObjectiveToinvestigatetheeffectofDNMT3AonDNAmethylationmodificationoftheSOX9gene.MethodsNormalchondrocytesandOAchondrocyteswerecultured.Themorphologyofchondrocyteswasobservedbymicroscope.ToluidinebluestainingandtypeⅡcollagenstainingwereusedtoidentifychondrocytes.DNMT3A,SOX9mRNAandproteinexpressionlevelsweredetectedbyqRT-PCRandWesternBlot.ThemethylationstatusofSOX9promoterwasdetectedbybisulfitesequencing.Dnmt3AwassilencedbylentivirustoobservethechangesofDNAmethylationmodificationofSOX9gene.ResultsNormalchondrocyteshadregularmorphology,largevolumeandlargenumber,whileOAchondrocyteshadirregularmorphology,smallvol...
