lncRNA在骨质疏松中作用的初步研究
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E-mail:sunqiang_cn@njmu.edu.cn

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R681

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南京市卫生科技发展专项资金项目(ZKX14032,ZKX19025)


Prelimnary study one the role of lncRNA in osteoporosis
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    摘要:

    目的:探究原发性骨质疏松(osteoporosis,OP)中差异基因及lncRNA RAD51⁃AS1的作用,为原发性骨质疏松的诊断及治疗提供新思路。方法:利用 GEO(Gene Expression Omnibus)数据库中的 GSE35956 芯片数据进行分析,使用 DEseq2 对 mRNA进行差异分析获得差异基因(differentially expressed gene,DEG),使用ClusterProfiler对差异分析的基因进行富集分析; 随后对芯片进行长链非编码RNA(long non⁃coding RNA,lncRNA)重注释以发掘差异lncRNA,运用实时定量PCR(qRT⁃PCR)对正常人及骨质疏松患者的骨髓间充质干细胞(human bone marrow mesenchymal stem cell,hBMSC)进行表达量验证。最后用 MTT、克隆形成和ALP染色观察RAD51⁃AS1对hBMSC增殖及成骨分化的影响。结果:共获得关于mRNA的1 440个DEG,其功能主要富集于受体配体活性和单核细胞分化等过程。同时也获得关于lncRNA的105个DEG,发现RAD51⁃AS1在骨质疏松患者中表达显著下调。沉默RAD51⁃AS1后发现hBMSC的增殖减少和ALP活性下降。结论:PI3K⁃Akt信号转导途径和单核细胞分化在骨质疏松中发挥重要作用。lncRNA RAD51⁃AS1可以调节骨髓间充质干细胞增殖及分化,以缓解骨质疏松进程。

    Abstract:

    Objective:To investigate the role of differential genes and lncRNA RAD51⁃AS1 in primary osteoporosis(OP),which provides new insights for the diagnosis and treatment of primary osteoporosis. Methods:The GSE35956 microarray data from the GEO database was used for analysis. Differentially expressed genes(DEG)were obtained by using the DEseq2,and the ClusterProfiler was used to find the enrichment of DEGs. The microarrays were then reannotated to discover the differential long ⁃ non coding RNA (lncRNA)in OP. The expression of RAD51⁃AS1 in hBMSC was verified by qRT⁃PCR. Finally,the effects of RAD51⁃AS1 on hBMSC proliferation and osteogenic differentiation were examined by MTT,colony formation,and ALP staining. Results:A total of 1 440 DEG was obtained about mRNAs in this study,whose functions were mainly enriched in processes such as receptor ⁃ ligand activity and monocyte differentiation. A total of 105 DEG regarding lncRNAs were also acquired. We found that RAD51 ⁃ AS1 expression was significantly downregulated in osteoporosis. The proliferation and ALP activity of hBMSC was reduced after silencing RAD51⁃AS1. Conclusions:The PI3K ⁃Akt signaling pathway and monocyte differentiation plays important roles in osteoporosis by bioinformatics. LncRNA RAD51 ⁃ AS1 regulates the proliferation and differentiation of bone marrow mesenchymal stem cells to ameliorate the progression of osteoporosisby cytological verification.

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李北辰,徐芳蓉,杨崇正,刘衡,孙强. lncRNA在骨质疏松中作用的初步研究[J].南京医科大学学报(自然科学版),2022,42(8):1100-1106

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  • 在线发布日期: 2022-08-07
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