Huang Guangxin, Kang Yan, Huang Zhiyu, Zhang Zhiqi, Meng Fangang, Chen Weishen, Fu Ming, Liao Weiming, Zhang Ziji
Cell Physiol Biochem. 2017;42(3):1037-1050. doi: 10.1159/000478751. Epub 2017 Jun 28.
BACKGROUND/AIMS: Long noncoding RNAs (lncRNAs) play important roles in stem cell differentiation. However, their role in osteogenesis of human adipose-derived stem cells (ASCs), a promising cell source for bone regeneration, remains unknown. Here, we investigated the expression profile and potential roles of lncRNAs in osteogenic differentiation of human ASCs.
Human ASCs were induced to differentiate into osteoblasts in vitro, and the expression profiles of lncRNAs and mRNAs in undifferentiated and osteogenic differentiated ASCs were obtained by microarray. Bioinformatics analyses including subgroup analysis, gene ontology analysis, pathway analysis and co-expression network analysis were performed. The function of lncRNA H19 was determined by in vitro knockdown and overexpression. Quantitative reverse transcription polymerase chain reaction was utilized to examine the expression of selected genes.
We identified 1,460 upregulated and 1,112 downregulated lncRNAs in osteogenic differentiated human ASCs as compared with those of undifferentiated cells (Fold change ≥ 2.0, P < 0.05). Among these, 94 antisense lncRNAs, 85 enhancer-like lncRNAs and 160 lincRNAs were further recognized. We used 12 lncRNAs and 157 mRNAs to comprise a coding-non-coding gene expression network. Additionally, silencing of H19 caused a significantly increase in expression of osteogenesis-related genes, including ALPL and RUNX2, while a decrease was observed after H19 overexpression.
This study revealed for the first time the global expression profile of lncRNAs involved in osteogenic differentiation of human ASCs and provided a foundation for future investigations of lncRNA regulation of human ASC osteogenesis.
背景/目的：长链非编码RNA（lncRNA）在干细胞分化中发挥重要作用。然而，它们在人脂肪来源干细胞（ASC）成骨过程中的作用尚不清楚，而ASC是一种很有前景的骨再生细胞来源。在此，我们研究了lncRNA在人ASC成骨分化中的表达谱及潜在作用。
体外诱导人ASC分化为成骨细胞，通过微阵列获得未分化及成骨分化ASC中lncRNA和mRNA的表达谱。进行了包括亚组分析、基因本体分析、通路分析和共表达网络分析在内的生物信息学分析。通过体外敲低和过表达确定lncRNA H19的功能。利用定量逆转录聚合酶链反应检测所选基因的表达。
与未分化细胞相比，我们在成骨分化的人ASC中鉴定出1460个上调的lncRNA和1112个下调的lncRNA（倍数变化≥2.0，P<0.05）。其中，进一步识别出94个反义lncRNA、85个增强子样lncRNA和160个长链间核糖核酸。我们使用12个lncRNA和157个mRNA构建了一个编码-非编码基因表达网络。此外，沉默H19导致成骨相关基因（包括碱性磷酸酶和RUNX2）的表达显著增加，而H19过表达后则观察到表达下降。
本研究首次揭示了参与人ASC成骨分化的lncRNA的整体表达谱，为未来研究lncRNA对人ASC成骨的调控提供了基础。
