Zhang Meng, Chen Delong, Zhang Fan, Zhang Gangyu, Wang Yueqi, Zhang Qingwen, He Wei, Wang Haibin, Chen Peng
The First School of Clinical Medicine, Guangzhou University of Chinese Medicine Guangzhou 510405, China.
Lingnan Medical Research Center, Guangzhou University of Chinese Medicine Guangzhou 510405, China.
Am J Transl Res. 2020 May 15;12(5):2136-2154. eCollection 2020.
Accumulating studies have demonstrated serum exosomal microRNAs (miRNAs) represent novel biomarkers for various diseases. In this study, we aimed to explore the feasibility of using serum exosomal miRNAs as novel serological biomarkers for steroid-induced osteonecrosis of femoral head (SONFH). We identified the characters of exosomes which were obtained from fresh serum of 5 systemic lupus erythematosus (SLE) patients without SONFH, 5 SLE patients with SONFH (SLE-SONFH) and 5 healthy ones. Comprehensive exosomal miRNA sequencing was performed to profile the differentially expressed miRNAs in the three groups. We then validated the expression levels of selected miRNAs by qRT-PCR. Furthermore, KEGG pathway, GO annotation, protein-protein interaction (PPI) network, module analysis and miRNAs-mRNAs interaction network were built to analyze the potential targets and mechanism. Sequencing data conveyed that hsa-miR-135b-5p, hsa-miR-150-5p, hsa-miR-509-3-5p, hsa-miR-514a-3p and hsa-miR-708-5p were significantly differentially expressed in the three groups. The results of qRT-PCR for the first time confirmed that the expression of hsa-miR-135b-5p was strikingly up-regulated in SLE-SONFH group which were consistent with miRNA sequencing results. In addition, bioinformatics analysis indicated that the enriched functions and pathways of the most differentially expressed miRNAs including Wnt, MAPK as well as Hippo signaling pathway. The top five hub genes (FGF2, PTEN, HACE1, VAMP2, and CBL) were part of module of the PPI network, which consisted of 713 nodes and 2191 edges. In conclusion, this study provides a novel and fundamental serum exosomal miRNAs profile of SONFH and hsa-miR-135b-5p may be identified as a unique diagnostic biomarker for SONFH.
越来越多的研究表明,血清外泌体微小RNA(miRNA)是多种疾病的新型生物标志物。在本研究中,我们旨在探讨使用血清外泌体miRNA作为类固醇诱导的股骨头坏死(SONFH)新型血清生物标志物的可行性。我们鉴定了从5例无SONFH的系统性红斑狼疮(SLE)患者、5例患有SONFH的SLE患者(SLE-SONFH)和5例健康者的新鲜血清中获得的外泌体特征。进行了全面的外泌体miRNA测序,以分析三组中差异表达的miRNA。然后,我们通过qRT-PCR验证了所选miRNA的表达水平。此外,构建了KEGG通路、GO注释、蛋白质-蛋白质相互作用(PPI)网络、模块分析和miRNAs-mRNAs相互作用网络,以分析潜在的靶点和机制。测序数据表明,hsa-miR-135b-5p、hsa-miR-150-5p、hsa-miR-509-3-5p、hsa-miR-514a-3p和hsa-miR-708-5p在三组中显著差异表达。qRT-PCR结果首次证实,hsa-miR-135b-5p在SLE-SONFH组中的表达显著上调,这与miRNA测序结果一致。此外,生物信息学分析表明,差异表达最显著的miRNA所富集的功能和通路包括Wnt、MAPK以及Hippo信号通路。前五个枢纽基因(FGF2、PTEN、HACE1、VAMP2和CBL)是PPI网络模块的一部分,该网络由713个节点和2191条边组成。总之,本研究提供了一种新型的、基础的SONFH血清外泌体miRNA图谱,hsa-miR-135b-5p可能被确定为SONFH的独特诊断生物标志物。