Abstract
Osteoarthritis (OA) is the most common chronic and degenerative joint disease. Although traditional OA medications can partially relieve pain, these medications cannot completely cure OA. Therefore, it is particularly important to find an effective treatment for OA. This study explored the function of long non-coding RNA (lncRNA)-colorectal neoplasia differentially expressed gene (CRNDE) in the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and the underlying molecular mechanism, aiming to develop a new treatment method for osteoarthritis. BMSCs were isolated from rat bone marrow using the gradient centrifugation method. And BMSC chondrogenic differentiation was induced with chondrogenic medium. The expression of lncRNA-CRNDE was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Silent information regulator factor 2-related enzyme 1 (SIRT1) and cartilage marker genes Aggrecan and collagen 2 (α1) protein expression were researched using western blot. Alcian blue staining was employed to examine the content of cartilage matrix proteoglycan glycosaminoglycan (GAG). The interaction between lncRNA-CRNDE and SIRT1 was detected by RNA pull-down and RNA immunoprecipitation (RIP) assay. Ubiquitination experiments were performed to measure the ubiquitination level of SIRT1. The combination between SMAD ubiquitination regulatory factor 2 (SMURF2) and SIRT1, as well as SRY-related high-mobility-group box 9 (SOX9) and collagen 2 (α1) promoter, was detected by Co-immunoprecipitation or ChIP. With the prolongation of induction time, the expression of lncRNA-CRNDE, SIRT1, cartilage marker genes Aggrecan and collagen 2 (α1) in BMSC osteogenic differentiation was gradually increased. Also, the content of cartilage matrix proteoglycan GAG was gradually elevated with the extension of the induction time. Further increase in the expression of SIRT1, cartilage marker genes Aggrecan and collagen 2 (α1) by overexpression of lncRNA-CRNDE also indicated elevated GAG content. RNA pull-down and RIP assay confirmed the binding between lncRNA-CRNDE and SIRT1. qRT-PCR and western blot showed that interference with lncRNA-CRNDE significantly inhibited the protein expression of SIRT1. BMSCs transfected with si-CRNDE increased ubiquitination levels of SIRT1 mediated by the E3 ligase SMURF2, leading to the reduced protein stability of SIRT1. However, overexpression of lncRNA-CRNDE increased the binding ability of SOX9 and collagen 2 (α1) promoter, which was reversed by the simultaneous transfection of CRNDE overexpression (pcDNA-CRNDE) and SIRT1 small interfering RNA (si-SIRT1). lncRNA-CRNDE regulates BMSC chondrogenic differentiation to promote cartilage repair in osteoarthritis through SIRT1/SOX9.
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References
Li MH, Xiao R, Li JB, Zhu Q (2017) Regenerative approaches for cartilage repair in the treatment of osteoarthritis. Osteoarthr Cartil 25(10):1577–1587
Pap T, Korb-Pap A (2015) Cartilage damage in osteoarthritis and rheumatoid arthritis—two unequal siblings. Nat Rev Rheumatol 11(10):606–615
Qi Y, Feng G, Yan W (2012) Mesenchymal stem cell-based treatment for cartilage defects in osteoarthritis. Mol Biol Rep 39(5):5683–5689
Tian Y, Xu Y, Fu Q, Chang M, Wang Y, Shang X, Wan C, Marymont JV, Dong Y (2015) Notch inhibits chondrogenic differentiation of mesenchymal progenitor cells by targeting Twist. Mol Cell Endocrinol 403:30–38
Li Z, Xu SF, Li DC, Sun Z, Zhang T, Lu JX, Wang Z (2014) Composite artificial semi-knee joint system. Eur Rev Med Pharmacol Sci 18(8):1229–1240
Richette P, Bardin T (2004) Structure-modifying agents for osteoarthritis: an update. Jt Bone Spine 71(1):18–23
Ham O, Lee CY, Kim R, Lee J, Oh S, Lee MY, Kim J, Hwang KC, Maeng LS, Chang W (2015) Therapeutic potential of differentiated mesenchymal stem cells for treatment of osteoarthritis. Int J Mol Sci 16(7):14961–14978
Koga H, Engebretsen L, Brinchmann JE, Muneta T, Sekiya I (2009) Mesenchymal stem cell-based therapy for cartilage repair: a review. Knee Surg Sports Traumatol Arthroscopy 17(11):1289–1297
Hollander AP, Dickinson SC, Kafienah W (2010) Stem cells and cartilage development: complexities of a simple tissue. Stem Cells (Dayton, OH) 28(11):1992–1996
Qu P, Wang L, Min Y, McKennett L, Keller JR, Lin PC (2016) Vav1 regulates mesenchymal stem cell differentiation decision between adipocyte and chondrocyte via SirT1. Stem Cells (Dayton, OH) 34(7):1934–1946
Dvir-Ginzberg M, Gagarina V, Lee EJ, Hall DJ (2008) Regulation of cartilage-specific gene expression in human chondrocytes by SirT1 and nicotinamide phosphoribosyltransferase. J Biol Chem 283(52):36300–36310
Simic P, Zainabadi K, Bell E, Sykes DB, Saez B, Lotinun S, Baron R, Scadden D, Schipani E, Guarente L (2013) SIRT1 regulates differentiation of mesenchymal stem cells by deacetylating β-catenin. EMBO Mol Med 5(3):430–440
Yang L, Froberg JE, Lee JT (2014) Long noncoding RNAs: fresh perspectives into the RNA world. Trends Biochem Sci 39(1):35–43
Shu T, He L, Wang X, Pang M, Yang B, Feng F, Wu Z, Liu C, Zhang S, Liu B et al (2019) Long noncoding RNA UCA1 promotes chondrogenic differentiation of human bone marrow mesenchymal stem cells via miRNA-145-5p/SMAD5 and miRNA-124-3p/SMAD4 axis. Biochem Biophys Res Commun 514(1):316–322
Barter MJ, Gomez R (2017) The long non-coding RNA ROCR contributes to SOX9 expression and chondrogenic differentiation of human mesenchymal stem cells. Development 144(24):4510–4521
Zhang L, Sun X, Chen S, Yang C, Shi B, Zhou L, Zhao J (2017) Long noncoding RNA DANCR regulates miR-1305-Smad 4 axis to promote chondrogenic differentiation of human synovium-derived mesenchymal stem cells. Biosci Rep 37(4):BSR20170347
Hu P, Sun F, Ran J, Wu L (2019) Identify CRNDE and LINC00152 as the key lncRNAs in age-related degeneration of articular cartilage through comprehensive and integrative analysis. PeerJ 7:e7024
Malynn BA, Ma A (2010) Ubiquitin makes its mark on immune regulation. Immunity 33(6):843–852
Yu L, Dong L, Li H, Liu Z, Luo Z, Duan G, Dai X, Lin Z (2020) Ubiquitination-mediated degradation of SIRT1 by SMURF2 suppresses CRC cell proliferation and tumorigenesis. Oncogene 39(22):4450–4464
Xiong H, Ni Z, He J, Jiang S, Li X, He J, Gong W, Zheng L, Chen S, Li B et al (2017) LncRNA HULC triggers autophagy via stabilizing Sirt1 and attenuates the chemosensitivity of HCC cells. Oncogene 36(25):3528–3540
Liu S, Zhang E, Yang M, Lu L (2014) Overexpression of Wnt11 promotes chondrogenic differentiation of bone marrow-derived mesenchymal stem cells in synergism with TGF-β. Mol Cell Biochem 390(1–2):123–131
Wang L, Yang J, Wang H, Wang W, Liang X (2020) Highly expressed ribosomal protein L34 predicts poor prognosis in acute myeloid leukemia and could be a potential therapy target. Aging Pathobiol Ther 2(1):32–37
Xing J, Liu H, Yang H, Chen R, Chen Y, Xu J (2014) Upregulation of Unc-51-like kinase 1 by nitric oxide stabilizes SIRT1, independent of autophagy. PLoS One 9(12):e116165
Baek D, Lee KM, Park KW, Suh JW, Choi SM, Park KH, Lee JW, Kim SH (2018) Inhibition of miR-449a promotes cartilage regeneration and prevents progression of osteoarthritis in in vivo rat models. Mol Ther Nucleic Acids 13:322–333
Zhang JJ, Fan LP (2019) Long non-coding RNA CRNDE enhances cervical cancer progression by suppressing PUMA expression. Biomed Pharmacother 117:108726
Li P, Pan X, Zheng Z, Sun Y, Han Y, Dong J (2020) LINC00271 inhibits epithelial-mesenchymal transition of papillary thyroid cancer cells by downregulating trefoil factor 3 expression. Aging Pathobiol Ther 2(2):78–85
Roman MG, Flores LC, Cunningham GM, Cheng C, Dube S, Allen C, Remmen HV, Bai Y, Hubbard GB, Saunders TL et al (2020) Thioredoxin overexpression in mitochondria showed minimum effects on aging and age-related diseases in male C57BL/6 mice. Aging Pathobiol Ther 2(1):20–31
Gelse K, Ekici AB, Cipa F, Swoboda B, Carl HD, Olk A, Hennig FF, Klinger P (2012) Molecular differentiation between osteophytic and articular cartilage—clues for a transient and permanent chondrocyte phenotype. Osteoarthr Cartil 20(2):162–171
Shimizu H, Yokoyama S, Asahara H (2007) Growth and differentiation of the developing limb bud from the perspective of chondrogenesis. Develop Growth Differ 49(6):449–454
Shen L, Chen L, Wang Y, Jiang X, Xia H, Zhuang Z (2015) Long noncoding RNA MALAT1 promotes brain metastasis by inducing epithelial-mesenchymal transition in lung cancer. J Neuro-Oncol 121(1):101–108
Wu P, Zuo X, Deng H, Liu X, Liu L, Ji A (2013) Roles of long noncoding RNAs in brain development, functional diversification and neurodegenerative diseases. Brain Res Bull 97:69–80
Ernst C, Morton CC (2013) Identification and function of long non-coding RNA. Front Cell Neurosci 7:168
Xie SC, Zhang JQ, Jiang XL, Hua YY, Xie SW, Qin YA, Yang YJ (2020) LncRNA CRNDE facilitates epigenetic suppression of CELF2 and LATS2 to promote proliferation, migration and chemoresistance in hepatocellular carcinoma. Cell Death Dis 11(8):676
Mulati M, Kobayashi Y, Takahashi A, Numata H, Saito M, Hiraoka Y, Ochi H, Sato S, Ezura Y, Yuasa M et al (2020) The long noncoding RNA Crnde regulates osteoblast proliferation through the Wnt/β-catenin signaling pathway in mice. Bone 130:115076
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Shi, C., Zheng, W. & Wang, J. lncRNA-CRNDE regulates BMSC chondrogenic differentiation and promotes cartilage repair in osteoarthritis through SIRT1/SOX9. Mol Cell Biochem 476, 1881–1890 (2021). https://doi.org/10.1007/s11010-020-04047-4
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DOI: https://doi.org/10.1007/s11010-020-04047-4