Identification of MicroRNAs by Microarray Analysis and Prediction of Target Genes Involved in Osteogenic Differentiation of Human Periodontal Ligament Stem Cells
Identification of MicroRNAs by Microarray Analysis and Prediction of Target Genes Involved in Osteogenic Differentiation of Human Periodontal Ligament Stem Cells
复制标题
微阵列分析鉴定MicroRNA并预测人牙周膜干细胞成骨分化相关靶基因
DOI：
10.1902/jop.2017.170079
复制
发表时间：
2017-10-01
影响因子：
4.3
通讯作者：
Fang, Fuchun
中科院分区：
文献类型：
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作者：
Hao, Yilin;Ge, Yihong;Fang, Fuchun
背景
microRNAs（miRNAs）在人牙周膜干细胞（hPDLSCs）成骨分化中的作用尚不清楚。在这项研究中，在hPDLSCs成骨分化的潜在分子机制进行了研究，使用miRNA分析。
方法
利用微阵列分析成骨分化过程中的miRNA表达谱。通过生物信息学方法预测出表达量变化至少2倍（P <0. 05）的miRNAs靶基因。通过定量逆转录-聚合酶链反应（qRT-PCR）验证了6种具有成骨相关靶基因的miRNA。
结果
116个miRNAs的表达被发现在骨诱导后改变，其中30个上调，86个下调。其中31个（26.7%）有成骨相关的靶基因。通过qRT-PCR验证了31种miRNA中的6种（miR-654- 3 p、miR-4288、miR-34 c-5 p、miR-218- 5 p、miR-663 a和miR-874- 3 p）表达水平的变化。
结论
在hPDLSC的成骨分化过程中观察到miRNA表达谱的显著改变。这些结果表明，miRNAs可能通过靶向成骨相关基因对这一过程产生调节作用。
BACKGROUND
The roles of microRNAs (miRNAs) in osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) remain largely unexplored. In this study, the underlying molecular mechanism of osteogenic differentiation in hPDLSCs is investigated using miRNA profiling.
METHODS
The miRNA expression profile during osteogenic differentiation was analyzed using a microarray. Target genes of miRNAs with at least two-fold change in expression (P <0.05) were predicted by bioinformatics. Six miRNAs with osteogenesis-related target genes were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
RESULTS
Expression of 116 miRNAs was found to be altered after osteoinduction, with 30 upregulated and 86 downregulated. Thirty-one of these miRNAs (26.7%) had osteogenesis-related target genes. Changes in expression levels of six of the 31 miRNAs (miR-654-3p, miR-4288, miR-34c-5p, miR-218-5p, miR-663a, and miR-874-3p) were validated by qRT-PCR.
CONCLUSIONS
Significant alterations in miRNA expression profiles were observed during osteogenic differentiation of hPDLSCs. These results imply that miRNAs may have regulatory effects on this process by targeting osteogenesis-related genes.