敲降人组织特异性lncRNA MEK6-AS1改善衰老间充质干细胞成骨分化能力  

Knockdown of human-specific lncRNA MEK6-AS1 improves osteogenic differentiation of senescent mesenchymal stem cells

作　　者：陈运华 李迪 赵晓妍 陈明 李红凌[1] 赵春华[1] CHEN Yunhua;LI Di;ZHAO Xiaoyan;CHEN Ming;LI Hongling;ZHAO Chunhua(Center for Excellence in Tissue Engineering,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,组织工程研究中心,北京100005

出　　处：《基础医学与临床》2025年第5期568-574,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(82474345,82471598);中国医学科学院医学与健康科技创新工程(2022-I2M-1-012);高等学校学科创新引智计划(B18007);全国重点实验室专项经费(2060204)。

摘　　要：目的旨在探讨一条新发现的人组织特异性长链非编码RNA(lncRNA)MEK6-AS1在改善人间充质干细胞(hMSCs)衰老后的成骨分化能力中的作用及分子机制。方法建立体外hMSCs复制性衰老细胞模型,利用RT-qPCR检测MEK6-AS1在衰老过程中的表达差异;采用慢病毒敲降技术抑制MEK6-AS1的表达,利用转录组测序技术分析MEK6-AS1对成骨相关转录组的影响;通过诱导细胞中成骨分化标志物的mRNA及蛋白质表达水平以及碱性磷酸酶(ALP)染色检测ALP活性,评估验证以MEK6-AS1为干预靶点对人衰老间充质干细胞成骨分化能力的改善效果。结果随着衰老的进展,hMSCs的成骨分化能力明显下降,MEK6-AS1的表达显著上调(P<0.0001)。在成骨诱导过程中,与衰老的对照组细胞相比,敲降MEK6-AS1的衰老细胞中成骨标志物的表达更高,诱导细胞的ALP活性更强(P<0.001)。结论敲降人组织特异性lncRNA MEK6-AS1可提升衰老MSCs的成骨分化能力,lncRNA MEK6-AS1可作为衰老相关骨质疏松防治的新靶点。Objective To investigate the role of a novel human-specific long noncoding RNA(lncRNA),MEK6-AS1,and its potential molecular mechanism in improving the osteogenic differentiation of senescent mesenchymal stem cells(MSCs).Methods An in vitro human MSCs replicative senescence model was established by sequential passaging,and then senescence was identified byβstaining and senescence-related gene expression.RT-qPCR was used to detect the expression of MEK6-AS1 during senescence and osteogenic differentiation of human MSCs(hMSCs);Lentiviral knockdown technology was used to regulate the expression of MEK6-AS1 in the MSCs replicative senescence model.Transcriptome sequencing technology was utilized to analyze the effects of MEK6-AS1 on the transcriptome of hMSCs especially on genes and pathways related to osteogenic differentiation.The effect of MEK6-AS1 as an intervention target at osteogenic differentiation of human senescent hMSCs was evaluated with alkaline phosphatase staining microscopy.PCR technology was used to detect osteogenic gene expression levels in cells.Results With aging process,the osteogenic differentiation ability of hMSCs decreased significantly while the expression level of MEK6-AS1 was enhanced(P<0.0001).Knockdown of MEK6-AS1 significantly enhanced the osteogenic differentiation of MSCs by the up-regulating expression of osteogenic markers and increasing mineralization capacity(P<0.001).Conclusions Knockdown of human-specific lncRNA MEK6-AS1 improves osteogenic differentiation of senescent MSCs,providing a new target and theoretical basis for the treatment of senescence-associated osteoporosis.

关 键 词：间充质干细胞(MSCs) 衰老 成骨分化 长链非编码RNA(lncRNA) MEK6-AS1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]