Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway (Retracted article. See vol. 14, 2023)
Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway (Retracted article. See vol. 14, 2023)
复制标题
长非编码 RNA HIF1A-AS2 通过 PI3K/Akt 信号通路通过 miR-665/IL6 轴促进脂肪源性干细胞 (ASC) 成骨分化
DOI：
10.1186/s13287-018-1082-z
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发表时间：
2018-12-13
影响因子：
7.5
通讯作者：
Wu, Qingkai
中科院分区：
文献类型：
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作者：
Wu, Ruoyu;Ruan, Jihao;Wu, Qingkai
本研究旨在通过PI3K/Akt信号通路探讨HIF1a-AS2/miR-665/IL6轴在脂肪来源干细胞成骨分化调控中的作用及其分子机制。利用R的BioConductor进行分析，基因集浓缩分析和京都百科全书的基因和基因组数据集用于识别上调和下调的信号通路。Cytoscape构建了特定的lncRNAs和mRNAs的共表达网络，TargetScan和Miranda预测了lncRNA、mRNAs和miRNA之间的结合位点。ASCs来源于人类脂肪组织，并通过流式细胞仪进行鉴定。茜素红和碱性磷酸酶(ALP)染色检测ASC细胞功能。通过RNAi、细胞转染、Western印迹和qRT-PCR等方法研究HIF1a-AS2/miR-665/IL6轴的分子机制。结果诱导的ASCs中高表达HIF1A-AS2和IL6，低表达miR-665。HIF1a-AS2和IL6促进成骨细胞标志物Runx2、Osterix和骨钙素的表达，并促进钙结节的形成和碱性磷酸酶的活性，而miR-665的作用相反。HIF1a-AS2直接靶向miR-665，而miR-665抑制IL6的表达。此外，HIF1a-AS2/miR-665/IL6调节轴激活了PI3K/Akt信号通路。结论LncRNA HIF1a-AS2可结合miR-665，上调IL6，激活PI3K/Akt信号通路，最终促进ASC成骨分化。
BackgroundThis study was aimed to investigate the role and specific molecular mechanism of HIF1A-AS2/miR-665/IL6 axis in regulating osteogenic differentiation of adipose-derived stem cells (ASCs) via the PI3K/Akt signaling pathway.MethodsRNAs' expression profile in normal/osteogenic differentiation-induced ASCs (osteogenic group) was from the Gene Expression Omnibus database. The analysis was carried out using Bioconductor of R. Gene Set Enrichment Analysis and Kyoto Encyclopedia of Genes and Genomes dataset were applied to identify up- and downregulated signaling pathways. Co-expression network of specific lncRNAs and mRNAs was structured by Cytoscape, while binding sites amongst lncRNA, mRNA, and miRNA were predicted by TargetScan and miRanda. ASCs were derived from human adipose tissue and were authenticated by flow cytometry. ASC cell function was surveyed by alizarin red and alkaline phosphatase (ALP) staining. Molecular mechanism of HIF1A-AS2/miR-665/IL6 axis was investigated by RNAi, cell transfection, western blot, and qRT-PCR. RNA target relationships were validated by dual-luciferase assay.ResultsHIF1A-AS2 and IL6 were highly expressed while miR-665 was lowly expressed in induced ASCs. HIF1A-AS2 and IL6 improved the expression level of osteoblast markers Runx2, Osterix, and Osteocalcin and also accelerated the formation of calcium nodule and ALP activity, yet miR-665 had opposite effects. HIF1A-AS2 directly targeted miR-665, whereas miR-665 repressed IL6 expression. Moreover, the HIF1A-AS2/miR-665/IL6 regulating axis activated the PI3K/Akt signaling pathway.ConclusionsLncRNA HIF1A-AS2 could sponge miR-665 and hence upregulate IL6, activate the PI3K/Akt signaling pathway, and ultimately promote ASC osteogenic differentiation.