补骨脂素诱导BMSCs成骨分化中的LncRNA表达谱分析  

Analysis of the expression profile of LncRNA in the differentiation of osteoblasts by psoralea-induced BMSCs

作　　者：杨锋[1] 李文雄[1] 康武林[1] 董博[1] 袁普卫[1] YANG Feng;LI Wenxiong;KANG Wulin;DONG Bo;YUAN Puwei(The Young Creation Team of Shannxi Universities, Shannxi Chinese Medical University, Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712046, China)

机构地区：[1]陕西中医药大学陕西高校青年创新团队,陕西中医药大学附属医院,陕西咸阳712046

出　　处：《中国骨质疏松杂志》2021年第3期385-391,共7页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金项目(81973889);陕西省重点科技创新团队项目(2013KCT-26)。

摘　　要：目的观察补骨脂诱导骨髓间充质干细胞(BMSCs)成骨分化过程中长链非编码RNA(lncRNA)的表达谱及相关靶基因和信号通路。方法取P2代BMSCs分为三组:空白组(含10%FBS的L-DMEM)、成骨诱导组(含10%FBS及成骨诱导剂的L-DMEM)、补骨脂素组(含10%FBS的L-DMEM及10μmol/L补骨脂素),同一环境不同诱导条件干预21 d;利用Agilent lncRNA芯片筛选出成骨诱导分化过程中表达差异2倍以上的lncRNA,并通过荧光定量PCR对芯片结果进行验证。选取筛选结果中表达差异较大的lncRNA进行GO和KEGG分析。结果成骨诱导分化21 d后持续表达超过2倍的lncRNA共有446个差异表达的lncRNAs。在BMSCs成骨分化过程中,共筛选出5个lncRNA显著上调,4个lncRNA显著下调。其中XR009483上调最显著,而XR007366下调最显著,经PCR验证与芯片结果相符。经GO分析富集度较高的为骨及软骨发育、干细胞分化等生物进程,以及胶原合成、骨化和钙化等生物功能。KEGG分析主要富集于15个生物学通路,其中富集评分较高的是TGF-beta、Wnt、NF-kappa B及Calcium等生物学通路。结论补骨脂素诱导BMSCs成骨分化过程中lncRNA表达谱发生显著变化,提示差异表达的lncRNA可能与BMSCs成骨分化密切相关,为研究补骨脂素促BMSCs成骨分化机制奠定了一定基础。Objective To observe the expression profile of long-chain non-coding RNA(lncRNA) and its related target genes and signal pathways during the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) induced by psoralen.Methods The second generation of BMSCs were divided into three groups:blank group(1-DMEM containing 10 % FBS)osteogenic induction group(L-DMEM containing 10 % FBS and osteogenic inducers),and psoralen group(L-DMEM containing10 μmol/L psoralen and 10 % FBS).After 21-day intervention,the difference of 2-fold variation of lncRNA in the process of osteogenic differentiation was screened using Agilent lncRNA chip.Real-time fluorescent quantitative PCR was used to verify the result of the chip.The lncRNA,which was highly differentially expressed in screening result was analyzed with Go and KEGG.Results There were 446 differentially expressed lncRNAs which expressed more than 2 times continuously after 21 days of osteogenic differentiation.In the process of osteogenic differentiation of BMSCs,5 up-regulated lncRNAs and 4 down-regulated lncRNAs were screened.Among those,XR009483 and XR007366 were most significantly up-regulated and down-regulated,respectively,and the result were confirmed with RT-PCR.The higher concentrations of GO were related to bone and cartilage development,stem cell differentiation,collagen synthesis,ossification,and calcification.Fifteen biological pathways was found in KEGG pathway analysis including TGF-beta,Wnt,NF-Kappa B,and calcium pathways.Conclusion The lncRNA expression profile of BMSCs changes significantly during osteogenic differentiation induced by psoralen,suggesting that the differential expression of lncRNA may be closely related to the osteogenic differentiation by BMSCs.

关 键 词：骨髓间充质干细胞 补骨脂素 成骨分化 长链非编码RNA 

分 类 号：R274.9[医药卫生—中西医结合]