Department of Orthopaedic Trauma, Honghui Hospital, Xi'an Jiaotong University, Xi'an, China.
Orthop Surg. 2021 Feb;13(1):285-295. doi: 10.1111/os.12866. Epub 2020 Dec 6.
To screen serum microRNAs (miRNAs) which could discriminate fracture status in postmenopausal women with or without diabetes.
The miRNA expression profile dataset GSE70318 was downloaded from Gene Expression Omnibus (GEO) database. This dataset composed of 74 samples, among these, 55 postmenopausal women was selected for bioinformatics analysis, including 19 osteoporotic fracture patients with type-2 diabetes, 19 osteoporotic fracture patients without type-2 diabetes, and 17 healthy control subjects. These samples were divided into two groups: fracture patients with diabetes vs healthy subjects (FH group) and fracture patients without diabetes vs healthy subjects (DFH group). Then, the differentially expressed miRNA (DEMs) in FH group and DFH group were respectively identified. The target genes of DEMs were predicted, followed by functional enrichment analysis. Furthermore, DEMs related to long non-coding RNAs (lncRNAs) were screened, and DEMs-lncRNA-target genes network was constructed. Subsequently, principal component analysis (PCA) of DEMs was performed to further explore the expression characteristics of the selected miRNAs in different types of fracture samples. Finally, the expression level of significant DEMs was calculated by quantitative real-time polymerase chain reaction (qPCR) to verify the accuracy of the results of bioinformatics analysis.
A total of 18 and 23 DEMs were identified in FH and DFH groups, respectively. Gene ontology (GO) analysis showed that genes in FH group were significantly enriched in regulation of transcription (GO: 0045449) and genes in DFH group were mainly enriched in cellular response to hormone stimulus (GO: 0032870). Meanwhile, pathway analysis indicated that genes in FH group were primarily enriched in T cell receptor signaling pathway (hsa04660) and genes in DFH group were mainly implicated in neurotrophin-signaling pathway (hsa04722). Moreover, the miRNA-lncRNA analysis revealed that miR-155-5p regulated by lncRNA MIR155HG was up-regulated in FH group; in addition, the miR-181c was significantly up-regulated and miR-375 was observably down-regulated in DFH group. Furthermore, PCA analysis suggested that the screened miRNAs were able to differentiate these two types of fractures in postmenopausal women. The miR-181c and miR-375 might be regarded as potential predictors for fracture, while miR-155-5p might be a candidate diagnostic biomarker for diabetic fracture. Finally, the results of qPCR were consistent with that of microarray data.
Overall, these three miRNAs might be regarded as potential diagnostic biomarkers to discriminate fracture status in postmenopausal women with and or without diabetes, and they served a putative role in the pathogenesis of these two diseases. However, these findings were only observed in serum samples and further clinical trials are urgently demanded to validate our results.
筛选能够区分绝经后女性伴或不伴糖尿病骨折状态的血清微小 RNA（miRNA）。
从基因表达综合数据库（GEO）下载 miRNA 表达谱数据集 GSE70318。该数据集由 74 个样本组成，其中选择了 55 名绝经后女性进行生物信息学分析，包括 19 名 2 型糖尿病骨质疏松性骨折患者、19 名无 2 型糖尿病骨质疏松性骨折患者和 17 名健康对照者。这些样本分为两组：伴糖尿病的骨折患者与健康受试者（FH 组）和不伴糖尿病的骨折患者与健康受试者（DFH 组）。然后，分别鉴定 FH 组和 DFH 组中差异表达的 miRNA（DEMs）。预测 DEMs 的靶基因，然后进行功能富集分析。此外，筛选与长链非编码 RNA（lncRNA）相关的 DEMs，并构建 DEMs-lncRNA-靶基因网络。随后，对 DEMs 进行主成分分析（PCA），以进一步探讨不同类型骨折样本中所选 miRNA 的表达特征。最后，通过实时定量聚合酶链反应（qPCR）计算显著 DEMs 的表达水平，以验证生物信息学分析结果的准确性。
FH 组和 DFH 组分别鉴定出 18 个和 23 个 DEMs。基因本体（GO）分析表明，FH 组中的基因主要富集在转录调控（GO：0045449），DFH 组中的基因主要富集在激素刺激的细胞反应（GO：0032870）。同时，通路分析表明，FH 组中的基因主要富集在 T 细胞受体信号通路（hsa04660），DFH 组中的基因主要涉及神经递质信号通路（hsa04722）。此外，miRNA-lncRNA 分析表明，FH 组中由 lncRNA MIR155HG 调控的 miR-155-5p 上调；此外，DFH 组中 miR-181c 显著上调，miR-375 明显下调。此外，PCA 分析表明，筛选出的 miRNAs 能够区分绝经后女性的这两种骨折。miR-181c 和 miR-375 可能是骨折的潜在预测因子，而 miR-155-5p 可能是糖尿病骨折的潜在诊断生物标志物。最后，qPCR 的结果与微阵列数据一致。
总体而言，这三种 miRNA 可能被视为区分绝经后女性伴或不伴糖尿病骨折状态的潜在诊断生物标志物，它们在这两种疾病的发病机制中可能发挥作用。然而，这些发现仅在血清样本中观察到，迫切需要进一步的临床试验来验证我们的结果。
