lncRNA-Xist在腰椎间盘退变髓核组织中的表达及临床意义  

Expression and clinical significance of lncRNA-Xist in lumbar disc degeneration nucleus pulposus tissue

作　　者：王凯利[1] Wang Kaili(China Academy of Traditional Chinese Medicine Guang'anmen Hospital South District,Beijing 100053,China)

机构地区：[1]中国中医科学院广安门医院南区,北京100053

出　　处：《国际生物医学工程杂志》2023年第6期536-542,共7页International Journal of Biomedical Engineering

摘　　要：目的探究长链非编码RNA(lncRNA)Xist在腰椎间盘退变(LIDD)髓核组织中的表达及临床意义。方法Trizol提取法提取LIDD髓核组织细胞及体外培养的髓核细胞总RNA,并通过实时荧光定量PCR验证LIDD髓核组织中lncRNA Xist的相对表达量;体外髓核组织细胞系中,通过转染si-lncRNA Xist构建敲低表达模型,实时荧光定量PCR鉴定敲低效率;EdU染色及流式细胞仪检测细胞增殖行为;Western Blot及流式细胞仪检测细胞凋亡。结果与对照组比较,LIDD髓核组织中lncRNA Xist的相对表达量升高(P<0.01)。与正常组比较,TNF-α组EdU阳性细胞数和S期细胞数占比降低(均P<0.01),凋亡细胞数增加(P<0.01),Bax蛋白表达水平升高(P<0.01),Bcl-2水平下降(P<0.05);与TNF-α组比较,TNF-α+siRNA-lncRNA Xist组EdU阳性细胞数和S期细胞数占比增加(均P<0.05),凋亡细胞数下调(P<0.05),Bax蛋白表达水平下降,Bcl-2水平升高。结论lncRNA Xist在LIDD患者髓核组织中高表达,且抑制髓核细胞增殖,促进凋亡,提示lncRNA Xist可作为LIDD的治疗靶点及潜在生物标志物。ObjectiveTo explore the expression and clinical significance of long non-coding RNA(lncRNA)Xist in the nucleus pulposus tissue of lumbar intervertebral disc degeneration(LIDD).MethodsThe Trizol extraction method was used to extract total RNA from LIDD nucleus pulposus tissue cells and in vitro cultured nucleus pulposus cells.The relative expression level of lncRNA Xist in LIDD nucleus pulposus tissue was verified through real-time fluorescence quantitative PCR.In vitro,a knockdown expression model was constructed by transfecting si-lncRNA Xist into a nucleus pulposus tissue cell line.The knockdown efficiency was identified using real-time fluorescence quantitative PCR.Cell proliferation behavior was detected using EdU staining and flow cytometry.Western Blot and flow cytometry were used to detect cell apoptosis.ResultsCompared with control group,lncRNA Xist expression in LIDD nucleus pulposus tissues was increased(P<0.01).Compared with normal group,the number of EdU positive cells and the number of S phase cells in TNF-αgroup were decreased(all P<0.01),cell apoptosis were increased(P<0.01),Bax protein expression was increased(P<0.01),and Bcl-2 protein expression was decreased(P<0.05).Compared with TNF-αgroup,the number of EdU positive cells and the number of S phase cells in TNF-α+siRNA-lncRNA Xist group were increased(all P<0.05),cell apoptosis were decreased(P<0.05),Bax protein expression was decreased,and Bcl-2 protein expression was increased.ConclusionslncRNA Xist is highly expressed in nucleus pulposus tissues of LIDD patients,inhibits the proliferation of nucleus pulposus cells,and promotes apoptosis,which suggests that lncRNA Xist may be used as a therapeutic target and biomarker of LIDD.

关 键 词：长链非编码RNA Xist 腰椎间盘退变 细胞增殖 细胞凋亡 

分 类 号：R681.53[医药卫生—骨科学]