人脂肪间充质干细胞成软骨分化过程中差异表达长链非编码RNA鉴定及功能  

Identification and function of differentially expressed long noncoding RNAs in the chondrogenesis of human adipose-derived mesenchymal stem cells

作　　者：孙红 邓进 彭国璇 庄勇 刘淼 宁旭 杨华 Sun Hong;Deng Jin;Peng Guoxuan;Zhuang Yong;Liu Miao;Ning Xu;Yang Hua(Department of Orthopedics,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Department of Emergency Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;School of Clinical Medicine,Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学附属医院骨科,贵州省贵阳市550004 [2]贵州医科大学附属医院急诊外科,贵州省贵阳市550004 [3]贵州医科大学临床医学院,贵州省贵阳市550004

出　　处：《中国组织工程研究》2023年第15期2325-2332,共8页Chinese Journal of Tissue Engineering Research

基　　金：贵州省卫生健康委科学技术基金(gzwjkj2020-1-120),项目负责人:孙红;贵州省卫生健康委科学技术基金(gzwkj2021-261),项目负责人:杨华;贵州省科技厅基础研究计划(黔科合基础-ZK[2021]一般391),项目负责人:杨华;贵州医科大学附属医院国家自然科学基金青年基金培育计划项目(gyfynsfc-2021-12),项目负责人:孙红;贵州省研究生科研基金立项课题(黔教合YJSKYJJ[2021]157),项目负责人:孙红;贵阳市科技局计划(筑科合同[2018]1-78),项目负责人:庄勇。

摘　　要：背景:人脂肪间充质干细胞成软骨分化过程受多种因素影响。研究证实,长链非编码RNAs在表观遗传调控、转录以及转录后水平调控等过程中扮演着重要角色,而目前其在人脂肪间充质干细胞成软骨分化过程中表达谱的改变以及调控作用鲜有报道。目的:探讨人脂肪间充质干细胞成软骨诱导分化过程中差异表达长链非编码RNAs及其功能。方法:以人脂肪间充质干细胞成软骨诱导分化0 d(未诱导组)、14 d(诱导组)细胞为研究对象,利用转录组测序技术筛选成软骨诱导分化前后差异表达倍数变化2倍及以上的长链非编码RNAs和mRNAs,并通过qRT-PCR对测序结果进行验证。采用生物信息学分析对差异表达基因行GO功能分析和KEGG通路富集分析,并筛选长链非编码RNAs邻近基因以及共表达基因。结果与结论:①与未诱导组相比,诱导组中显著差异表达的长链非编码RNAs共有816条,mRNAs共有5138条;②GO功能和KEGG信号通路分析发现,差异表达基因富集的主要生物学过程包括细胞进程、生物调节和代谢过程等;主要通路包括黏附斑激酶、胰岛素信号通路、Wnt信号通路等;③对差异表达长链非编码RNAs行邻近基因和共表达基因分析发现,部分长链非编码RNAs如SNHG16、XLOC_003886可能在脂肪间充质干细胞成软骨分化和软骨退变中发挥重要作用;④结果表明,长链非编码RNA的表达谱在脂肪间充质干细胞诱导成软骨分化过程中发生了明显改变;差异表达长链非编码RNAs的生物学功能可能与邻近基因和共表达基因功能密切相关,从而调控脂肪间充质干细胞成软骨分化。然而,其具体调控作用和分子机制有待实验进一步证实。BACKGROUND:The chondrogenic differentiation of human adipose-derived mesenchymal stem cells is affected by many factors.Studies have confirmed that long noncoding RNAs(lncRNAs)play important roles in epigenetic regulation,transcription and post-transcriptional regulation.However,there are few reports on the changes of its expression profile and its regulatory role during the chondrogenic differentiation of human adipose-derived mesenchymal stem cells.OBJECTIVE:To preliminarily analyze the differentially expressed lncRNAs and their potential functions during the chondrogenesis of human adipose-derived mesenchymal stem cells.METHODS:The human adipose-derived mesenchymal stem cells at 0 day(undifferentiated group)and 14 days(differentiated group)after chondrogenic differentiation were used as research objects.RNA-seq was used to screen lncRNAs and messenger RNAs(mRNAs)with expressed fold change larger than twice between groups.Quantitative real time PCR(qRT-PCR)was used to verify the sequencing results.Bioinformatics analyses including GO function analysis and KEGG pathway analysis were performed for differentially expressed genes to screen adjacent genes and co-expressed genes of lncRNAs.RESULTS AND CONCLUSION:(1)Compared with the undifferentiated group,a total of 816 lncRNAs and 5138 mRNAs were significantly differentially expressed in the differentiated group.(2)GO function and KEGG signaling pathway analysis showed that differentially expressed genes were enriched in several biological processes including cellular processes,biological regulation,and metabolic processes,and multiple pathways including focal adhesion kinase,insulin signaling pathway,and Wnt signaling pathway.(3)Analysis results of adjacent genes and co-expressed genes of differentially expressed lncRNAs indicated that some lncRNAs such as SNHG16 and XLOC_003886 might play critical roles in the chondrogenic differentiation of adipose-derived mesenchymal stem cells and cartilage degeneration.(4)These findings suggest that the expression profiles of

关 键 词：长链非编码RNA 脂肪间充质干细胞 成软骨分化 转录组测序 生物信息学分析 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R681.3