Department of Oral and Maxillofacial Surgery, Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital), Guangzhou, Guangdong 510280, P.R. China.
Department of Endodontics, Stomatological Hospital, Southern Medical University (Guangdong Provincial Stomatological Hospital), Guangzhou, Guangdong 510280, P.R. China.
Mol Med Rep. 2019 Jan;19(1):103-114. doi: 10.3892/mmr.2018.9674. Epub 2018 Nov 20.
Long non‑coding RNAs (lncRNAs) are a specific group of RNA molecules that do not encode proteins. They have been shown to serve important regulatory functions in various biological and cell differentiation processes. However, the potential functions and regulatory mechanisms of lncRNAs that are associated with the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) remain to be elucidated. The present study aimed to investigate lncRNAs that are differentially expressed during the osteogenic differentiation of hBMSCs, along with the potential functions of those lncRNAs. To this end, three groups of hBMSCs were stimulated to undergo osteogenic differentiation for 7 days. Known lncRNAs, unknown lncRNAs and mRNAs that demonstrated differential expression prior to and following the osteogenic differentiation of hBMSCs were screened using lncRNA high‑throughput sequencing. In addition, 12 lncRNAs were selected for reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) validation of the accuracy of the sequencing results. The potential functions and possible targets of the differentially expressed lncRNAs were analyzed using bioinformatics technologies (gene ontology, Kyoto Encyclopedia of Genes and Genomes and gene co‑expression network analysis). In total, 64 lncRNAs were differentially expressed by at least two‑fold in hBMSCs prior to and following osteogenic differentiation; these included seven known lncRNAs (two upregulated and five downregulated lncRNAs) and 57 unknown lncRNAs (35 upregulated and 22 downregulated lncRNAs). In addition, 409 mRNAs (257 upregulated and 152 downregulated mRNAs) were differentially expressed by at least two‑fold. The RT‑qPCR results obtained for 12 selected differentially expressed lncRNAs were consistent with the sequencing results. The gene co‑expression network analysis of lncRNAs and mRNAs demonstrated that four lncRNAs (ENSG00000238042, lnc_1269, lnc_1369 and lnc_1708) may serve important roles in the osteogenic differentiation of hBMSCs. In conclusion, during the osteogenic differentiation of hBMSCs, the lncRNA expression profile changed significantly; certain of the observed differentially expressed lncRNAs may be derived from protein‑coding genes and may serve important roles in osteogenic differentiation.
长非编码 RNA（lncRNA）是一组特定的 RNA 分子，它们不编码蛋白质。已经证明它们在各种生物和细胞分化过程中发挥着重要的调节功能。然而，lncRNA 与人类骨髓间充质干细胞（hBMSC）成骨分化相关的潜在功能和调节机制仍有待阐明。本研究旨在探讨在 hBMSC 成骨分化过程中差异表达的 lncRNA 及其潜在功能。为此，将三组 hBMSC 刺激进行 7 天的成骨分化。使用 lncRNA 高通量测序筛选 hBMSC 成骨分化前后差异表达的已知 lncRNA、未知 lncRNA 和 mRNAs。此外，选择 12 个 lncRNA 进行逆转录-定量聚合酶链反应（RT-qPCR）验证测序结果的准确性。使用生物信息学技术（基因本体、京都基因与基因组百科全书和基因共表达网络分析）分析差异表达 lncRNA 的潜在功能和可能的靶标。总共在 hBMSC 成骨分化前后有 64 个 lncRNA 差异表达至少两倍，包括七个已知 lncRNA（两个上调和五个下调 lncRNA）和 57 个未知 lncRNA（35 个上调和 22 个下调 lncRNA）。此外，有 409 个 mRNAs（257 个上调和 152 个下调 mRNAs）差异表达至少两倍。为 12 个选定的差异表达 lncRNA 进行的 RT-qPCR 结果与测序结果一致。lncRNA 和 mRNAs 的基因共表达网络分析表明，四个 lncRNA（ENSG00000238042、lnc_1269、lnc_1369 和 lnc_1708）可能在 hBMSC 成骨分化中发挥重要作用。总之，在 hBMSC 成骨分化过程中，lncRNA 表达谱发生了显著变化；观察到的某些差异表达 lncRNA 可能来自蛋白质编码基因，并可能在成骨分化中发挥重要作用。
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