Potential functions of long non‑coding RNAs in the osteogenic differentiation of human bone marrow mesenchymal stem cells

Sun,Xiang; Jia,Bo; Qiu,Xiao‑Ling; Chu,Hong‑Xing; Zhang,Zhao‑Qiang; Wang,Zhi‑Ping; Zhao,Jian‑Jiang

doi:10.3892/mmr.2018.9674

Potential functions of long non‑coding RNAs in the osteogenic differentiation of human bone marrow mesenchymal stem cells

Authors:
- Xiang Sun
- Bo Jia
- Xiao‑Ling Qiu
- Hong‑Xing Chu
- Zhao‑Qiang Zhang
- Zhi‑Ping Wang
- Jian‑Jiang Zhao
View Affiliations

Affiliations: 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
Published online on: November 20, 2018 https://doi.org/10.3892/mmr.2018.9674
Pages: 103-114
Copyright: © Sun et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

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.

View Figures

View References

1	Kinoshita Y and Maeda H: Recent developments of functional scaffolds for craniomaxillofacial bone tissue engineering applications. ScientificWorldJournal. 2013:8631572013. View Article : Google Scholar : PubMed/NCBI
2	Frith JE, Thomson B and Genever PG: Dynamic three-dimensional culture methods enhance mesenchymal stem cell properties and increase therapeutic potential. Tissue Eng Part C Methods. 16:735–749. 2010. View Article : Google Scholar : PubMed/NCBI
3	Zhong ZN, Zhu SF, Yuan AD, Lu GH, He ZY, Fa ZQ and Li WH: Potential of placenta-derived mesenchymal stem cells as seed cells for bone tissue engineering: Preliminary study of osteoblastic differentiation and immunogenicity. Orthopedics. 35:779–788. 2012. View Article : Google Scholar : PubMed/NCBI
4	Yan B and Wang Z: Long noncoding RNA: Its physiological and pathological roles. DNA Cell Biol. 31 Suppl 1:S34–S41. 2012. View Article : Google Scholar : PubMed/NCBI
5	Mercer TR, Dinger ME and Mattick JS: Long non-coding RNAs: Insights into functions. Nat Rev Genet. 10:155–159. 2009. View Article : Google Scholar : PubMed/NCBI
6	Luo S, Lu JY, Liu L, Yin Y, Chen C, Han X, Wu B, Xu R, Liu W, Yan P, et al: Divergent lncRNAs regulate gene expression and lineage differentiation in pluripotent cells. Cell Stem Cell. 18:637–652. 2016. View Article : Google Scholar : PubMed/NCBI
7	Takeda K, Ichijo H, Fujii M, Mochida Y, Saitoh M, Nishitoh H, Sampath TK and Miyazono K: Identification of a novel bone morphogenetic protein-responsive gene that may function as a noncoding RNA. J Biol Chem. 273:17079–17085. 1998. View Article : Google Scholar : PubMed/NCBI
8	Zhu L and Xu PC: Downregulated LncRNA-ANCR promotes osteoblast differentiation by targeting EZH2 and regulating Runx2 expression. Biochem Biophys Res Commun. 432:612–617. 2013. View Article : Google Scholar : PubMed/NCBI
9	Zuo C, Wang Z, Lu H, Dai Z, Liu X and Cui L: Expression profiling of lncRNAs in C3H10T1/2 mesenchymal stem cells undergoing early osteoblast differentiation. Mol Med Rep. 8:463–467. 2013. View Article : Google Scholar : PubMed/NCBI
10	Su JP, Tan LY, Garland SM, Tabrizi SN, Mokany E, Walker S, Bradshaw CS, Read T and Murray GL: Evaluation of the SpeeDx ResistancePlus MG diagnostic test for mycoplasma genitalium on the applied biosystems 7500 fast quantitative PCR platform. J Clin Microbiol. 56:e01245–17. 2017. View Article : Google Scholar : PubMed/NCBI
11	Okamoto H, Matsumi Y, Hoshikawa Y, Takubo K, Ryoke K and Shiota G: Involvement of microRNAs in regulation of osteoblastic differentiation in mouse induced pluripotent stem Cells. PLoS One. 7:e438002012. View Article : Google Scholar : PubMed/NCBI
12	Panetta NJ, Gupta DM, Quarto N and Longaker MT: Mesenchymal cells for skeletal tissue engineering. Panminerva Med. 51:25–41. 2009.PubMed/NCBI
13	Wang KC and Chang HY: Molecular mechanisms of long noncoding RNAs. Mol Cell. 43:904–914. 2011. View Article : Google Scholar : PubMed/NCBI
14	Yang L, Duff MO, Graveley BR, Carmichael GG and Chen LL: Genomewide characterization of non-polyadenylated RNAs. Genome Biol. 12:R162011. View Article : Google Scholar : PubMed/NCBI
15	Niazi F and Valadkhan S: Computational analysis of functional long noncoding RNAs reveals lack of peptide-coding capacity and parallels with 3′ UTRs. RNA. 18:825–843. 2012. View Article : Google Scholar : PubMed/NCBI
16	Ng SY, Johnson R and Stanton LW: Human long non-coding RNAs promote pluripotency and neuronal differentiation by association with chromatin modifiers and transcription factors. EMBO J. 31:522–533. 2012. View Article : Google Scholar : PubMed/NCBI
17	Sunwoo H, Dinger ME, Wilusz JE, Amaral PP, Mattick JS and Spector DL: MEN epsilon/beta nuclear-retained non-coding RNAs are upregulated upon muscle differentiation and are essential components of paraspeckles. Genome Res. 19:347–359. 2009. View Article : Google Scholar : PubMed/NCBI
18	Cesana M, Cacchiarelli D, Legnini I, Santini T, Sthandier O, Chinappi M, Tramontano A and Bozzoni I: A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA. Cell. 147:358–369. 2011. View Article : Google Scholar : PubMed/NCBI
19	Kretz M, Siprashvili Z, Chu C, Webster DE, Zehnder A, Qu K, Lee CS, Flockhart RJ, Groff AF, Chow J, et al: Control of somatic tissue differentiation by the long non-coding RNA TINCR. Nature. 493:231–235. 2013. View Article : Google Scholar : PubMed/NCBI
20	Kikuchi K, Fukuda M, Ito T, Inoue M, Yokoi T, Chiku S, Mitsuyama T, Asai K, Hirose T and Aizawa Y: Transcripts of unknown function in multiple-signaling pathways involved in human stem cell differentiation. Nucleic Acids Res. 37:4987–5000. 2009. View Article : Google Scholar : PubMed/NCBI
21	Jia Q, Jiang W and Ni L: Downregulated non-coding RNA (lncRNA-ANCR) promotes osteogenic differentiation of periodontal ligament stem cells. Arch Oral Biol. 60:234–241. 2015. View Article : Google Scholar : PubMed/NCBI
22	Cao B, Liu N and Wang W: High glucose prevents osteogenic differentiation of mesenchymal stem cells via lncRNA AK028326/CXCL13 pathway. Biomed Pharmacother. 84:544–551. 2016. View Article : Google Scholar : PubMed/NCBI
23	Li Z, Jin C, Chen S, Zheng Y, Huang Y, Jia L, Ge W and Zhou Y: Long non-coding RNA MEG3 inhibits adipogenesis and promotes osteogenesis of human adipose-derived mesenchymal stem cells via miR-140-5p. Mol Cell Biochem. 433:51–60. 2017. View Article : Google Scholar : PubMed/NCBI
24	Wang L, Wu F, Song Y, Li X, Wu Q, Duan Y and Jin Z: Long noncoding RNA associated with periodontitis interacts with miR-182 to upregulate osteogenic differentiation in periodontal mesenchymal stem cells of periodontitis patients. Cell Death Dis. 7:e23272016. View Article : Google Scholar : PubMed/NCBI
25	Hong IS, Lee HY, Choi SW, Kim HS, Yu KR, Seo Y, Jung JW and Kang KS: The effects of hedgehog on RNA binding protein Msi1 during the osteogenic differentiation of human cord blood-derived mesenchymal stem cells. Bone. 56:416–425. 2013. View Article : Google Scholar : PubMed/NCBI
26	Kovacevic A, Hammer A, Stadelmeyer E, Windischhofer W, Sundl M, Ray A, Schweighofer N, Friedl G, Windhager R, Sattler W and Malle E: Expression of serum amyloid A transcripts in human bone tissues, differentiated osteoblast-like stem cells and human osteosarcoma cell lines. J Cell Biochem. 103:994–1004. 2008. View Article : Google Scholar : PubMed/NCBI
27	Granéli C, Thorfve A, Ruetschi U, Brisby H, Thomsen P, Lindahl A and Karlsson C: Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach. Stem Cell Res. 12:153–165. 2014. View Article : Google Scholar : PubMed/NCBI
28	Abdallah BM, Ditzel N, Mahmood A, Isa A, Traustadottir GA, Schilling AF, Ruiz-Hidalgo MJ, Laborda J, Amling M and Kassem M: DLK1 is a novel regulator of bone mass that mediates estrogen deficiency-induced bone loss in mice. J Bone Miner Res. 26:1457–1471. 2011. View Article : Google Scholar : PubMed/NCBI