Abstract
Strontium is a drug with the bone formation and anti-resorption effects on bone. The underlying mechanisms for the dual effect of strontium on bone metabolism, especially for the anti-resorption effects remain unknown. Thus, we aim to investigate the mechanisms of effects of strontium on osteoclastogenesis. Firstly, we found that strontium decreased the levels of important biomarkers of receptor activator of nuclear factor kappa-B ligand (RANKL) which induced osteoclast differentiation, indicating that strontium might directly inhibit osteoclast differentiation. Next, we revealed that strontium enhanced Low Density Lipoprotein Receptor-Related Protein 6 (LRP6)/β-catenin/osteoprotegerin (OPG) signaling pathway in MC3T3-E1 cells. The signaling pathway may negatively regulate osteoclastogenesis. Thus, strontium indirectly inhibited RANKL induced osteoclast differentiation. Finally, we revealed that OPG was targeted by miR-181d-5p as determined by luciferase reporter assay and downregulated by miR-181d-5p at both mRNA and protein levels as determined by western blot.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- RANKL:
-
nuclear factor kappa-B ligand
- LRP6:
-
Low Density Lipoprotein Receptor-Related Protein 6
- OPG:
-
osteoprotegerin
- RBPJ:
-
Recombining binding protein suppressor of hairless
- HES:
-
hairy and enhancer of split
- HEY:
-
hairy/enhancer-of-split related with YRPW motif
- miRNAs:
-
MicroRNAs
- 3′UTR:
-
3′-untranslated region
- CALCR:
-
calcitonin receptor
- MMP9:
-
matrix metallopeptidase 9
- c-Fos:
-
FBJ osteosarcoma oncogene
- TRAP:
-
tartrate-resistant acid phosphatase
- CTSK:
-
cathepsin K
References
Bonnelye E, Chabadel A, Saltel F, Jurdic P (2008) Dual effect of strontium ranelate: stimulation of osteoblast differentiation and inhibition of osteoclast formation and resorption in vitro. Bone 42(1):129–138. https://doi.org/10.1016/j.bone.2007.08.043
Buehler J, Chappuis P, Saffar JL, Tsouderos Y, Vignery A (2001) Strontium ranelate inhibits bone resorption while maintaining bone formation in alveolar bone in monkeys (Macaca fascicularis). Bone 29(2):176–179. https://doi.org/10.1016/S8756-3282(01)00484-7
Burgers TA, Williams BO (2013) Regulation of Wnt/beta-catenin signaling within and from osteocytes. Bone 54(2):244–249. https://doi.org/10.1016/j.bone.2013.02.022
Cai T, Sun DQ, Duan Y, Wen P, Dai CS, Yang JW, He WC (2016) WNT/beta-catenin signaling promotes VSMCs to osteogenic transdifferentiation and calcification through directly modulating Runx2 gene expression. Exp Cell Res 345(2):206–217. https://doi.org/10.1016/j.yexcr.2016.06.007
Capaccione KM, Pine SR (2013) The notch signaling pathway as a mediator of tumor survival. Carcinogenesis 34(7):1420–1430. https://doi.org/10.1093/carcin/bgt127
Caudrillier A, Hurtel-Lemaire AS, Wattel A, Cournarie F, Godin C, Petit L et al (2010) Strontium ranelate decreases receptor activator of nuclear factor-KappaB ligand-induced osteoclastic differentiation in vitro: involvement of the calcium-sensing receptor. Mol Pharmacol 78(4):569–576. https://doi.org/10.1124/mol.109.063347
Chen JQ, Long FX (2013) Beta-catenin promotes bone formation and suppresses bone resorption in postnatal growing mice. J Bone Miner Res 28(5):1160–1169. https://doi.org/10.1002/jbmr.1834
Chen JQ, Tu XL, Esen E, Joeng KS, Lin CX, Arbeit JM et al (2014) WNT7B promotes bone formation in part through mTORC1. PLoS Genet 10(1). https://doi.org/10.1371/journal.pgen.1004145
Chen C, Qin Y, Fang JP, Ni XY, Yao J, Wang HY, Ding K (2015) WSS25, a sulfated polysaccharide, inhibits RANKL-induced mouse osteoclast formation by blocking SMAD/ID1 signaling. Acta Pharmacol Sin 36(9):1053–1064. https://doi.org/10.1038/aps.2015.65
Cheng CP, Sheu MJ, Sytwu HK, Chang DM (2013) Decoy receptor 3 suppresses RANKL-induced osteoclastogenesis via down-regulating NFATc1 and enhancing cell apoptosis. Rheumatology (Oxford) 52(4):609–622. https://doi.org/10.1093/rheumatology/kes343
Deregowski V, Gazzerro E, Priest L, Rydziel S, Canalis E (2006) Notch 1 overexpression inhibits osteoblastogenesis by suppressing Wnt/beta-catenin but not bone morphogenetic protein signaling. J Biol Chem 281(10):6203–6210. https://doi.org/10.1074/jbc.M508370200
Dige MS, Ahlawat SPS, Bhusan B, Kumar P, Chauhan A, Inamdar B et al (2013) Dissecting the partial genomic region of Cxcr to correlate with Cmt in Vrindavani cattle. Indian J Anim Res 47(4):335–339.
Fromigue O, Hay E, Barbara A, Marie PJ (2010) Essential role of nuclear factor of activated T cells (NFAT)-mediated Wnt signaling in osteoblast differentiation induced by strontium ranelate. J Biol Chem 285(33):25251–25258. https://doi.org/10.1074/jbc.M110.110502
Hurtel-Lemaire AS, Mentaverri R, Caudrillier A, Cournarie F, Wattel A, Kamel S et al (2009) THE calcium-sensing receptor is involved in strontium Ranelate-induced osteoclast apoptosis NEW INSIGHTS INTO THE ASSOCIATED SIGNALING PATHWAYS. J Biol Chem 284(1):575–584. https://doi.org/10.1074/jbc.M801668200
Jin HT, Wang BL, Li J, Xie WQ, Mao Q, Li S et al (2015) Anti-DKK1 antibody promotes bone fracture healing through activation of beta-catenin signaling. Bone 71:63–75. https://doi.org/10.1016/j.bone.2014.07.039
Jovanovic M, Hengartner MO (2006) miRNAs and apoptosis: RNAs to die for. Oncogene 25(46):6176–6187. https://doi.org/10.1038/sj.onc.1209912
Lee YS, Kim HK, Chung SM, Kim KS, Dutta A (2005) Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the downregulation of putative targets during differentiation. J Biol Chem 280(17):16635–16641. https://doi.org/10.1074/jbc.M412247200
Li SF, Liu B, Zhang LM, Rong LM (2014) Amyloid beta peptide is elevated in osteoporotic bone tissues and enhances osteoclast function. Bone 61:164–175. https://doi.org/10.1016/j.bone.2014.01.010
Lumetti S, Manfredi E, Ferraris S, Spriano S, Passeri G, Ghiacci G et al (2016) The response of osteoblastic MC3T3-E1 cells to micro- and nano-textured, hydrophilic and bioactive titanium surfaces. J Mater Sci Mater Med 27(4):68. https://doi.org/10.1007/s10856-016-5678-5
Ma J, Liu YL, Hu YY, Wei YN, Zhao XC, Dong GY et al (2013) Disruption of the transcription factor RBP-J results in osteopenia attributable to attenuated osteoclast differentiation. Mol Biol Rep 40(3):2097–2105. https://doi.org/10.1007/s11033-012-2268-6
Matsuzaki E, Hiratsuka S, Hamachi T, Takahashi-Yanaga F, Hashimoto Y, Higashi K et al (2013) Sphingosine-1-phosphate promotes the nuclear translocation of beta-catenin and thereby induces osteoprotegerin gene expression in osteoblast-like cell lines. Bone 55(2):315–324. https://doi.org/10.1016/j.bone.2013.04.008
Mi BG, Xiong W, Xu N, Guan HF, Fang Z, Liao H et al (2017) Strontium-loaded titania nanotube arrays repress osteoclast differentiation through multiple signalling pathways: in vitro and in vivo studies. Sci Rep 7. https://doi.org/10.1038/s41598-017-02491-9
Muguruma Y, Hozumi K, Warita H, Yahata T, Uno T, Ito M, Ando K (2016) Maintenance of bone homeostasis by DLL1-mediated notch signaling. J Cell Physiol. https://doi.org/10.1002/jcp.25647
Peng S, Liu XS, Huang S, Li Z, Pan H, Zhen W et al (2011a) The cross-talk between osteoclasts and osteoblasts in response to strontium treatment: involvement of osteoprotegerin. Bone 49(6):1290–1298. https://doi.org/10.1016/j.bone.2011.08.031
Peng S, Liu XS, Zhou G, Li Z, Luk KD, Guo XE, Lu WW (2011b) Osteoprotegerin deficiency attenuates strontium-mediated inhibition of osteoclastogenesis and bone resorption. J Bone Miner Res 26(6):1272–1282. https://doi.org/10.1002/jbmr.325
Riddle RC, Diegel CR, Leslie JM, Van Koevering KK, Faugere MC, Clemens TL, Williams BO (2013) Lrp5 and Lrp6 exert overlapping functions in osteoblasts during postnatal bone acquisition. PLoS One 8(5). https://doi.org/10.1371/journal.pone.0063323
Roy M, Bose S (2012) Osteoclastogenesis and osteoclastic resorption of tricalcium phosphate: effect of strontium and magnesium doping. J Biomed Mater Res A 100a(9):2450–2461. https://doi.org/10.1002/jbm.a.34181
Roy M, Fielding G, Bandyopadhyay A, Bose S (2013) Effects of zinc and strontium substitution in Tricalcium phosphate on osteoclast differentiation and resorption. Biomater Sci 1(1). https://doi.org/10.1039/C2BM00012A
Su R, Lin HS, Zhang XH, Yin XL, Ning HM, Liu B et al (2015) MiR-181 family: regulators of myeloid differentiation and acute myeloid leukemia as well as potential therapeutic targets. Oncogene 34(25):3226–3239. https://doi.org/10.1038/onc.2014.274
Sun T, Leung F, Lu WW (2016) MiR-9-5p, miR-675-5p and miR-138-5p damages the strontium and LRP5-mediated skeletal cell proliferation, differentiation, and adhesion. Int J Mol Sci 17(2):236. https://doi.org/10.3390/ijms17020236
Sun T, Cheung KSC, Liu ZL, Leung F, Lu WW (2017a) Matrix metallopeptidase 9 targeted by hsa-miR-494 promotes silybin-inhibited osteosarcoma. Mol Carcinog. https://doi.org/10.1002/mc.22753
Sun T, Li CT, Xiong L, Ning Z, Leung F, Peng S, Lu WW (2017b) miR-375-3p negatively regulates osteogenesis by targeting and decreasing the expression levels of LRP5 and beta-catenin. PLoS One 12(2):e0171281. https://doi.org/10.1371/journal.pone.0171281
Takahashi N, Sasaki T, Tsouderos Y, Suda T (2003) S 12911-2 inhibits osteoclastic bone resorption in vitro. J Bone Miner Res 18(6):1082–1087. https://doi.org/10.1359/jbmr.2003.18.6.1082
Takai H, Kanematsu M, Yano K, Tsuda E, Higashio K, Ikeda K et al (1998) Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastogenesis inhibitory factor by bone marrow stromal cells. J Biol Chem 273(42):27091–27096
Tezuka K, Yasuda M, Watanabe N, Morimura N, Kuroda K, Miyatani S, Hozumi N (2002) Stimulation of osteoblastic cell differentiation by notch. J Bone Miner Res 17(2):231–239. https://doi.org/10.1359/jbmr.2002.17.2.231
Theoleyre S, Wittrant Y, Tat SK, Fortun Y, Redini F, Heymann D (2004) The molecular triad OPG/RANK/RANKL: involvement in the orchestration of pathophysiological bone remodeling. Cytokine Growth Factor Rev 15(6):457–475. https://doi.org/10.1016/j.cytogfr.2004.06.004
Wang B, Hsu SH, Majumder S, Kutay H, Huang W, Jacob ST, Ghoshal K (2010) TGFbeta-mediated upregulation of hepatic miR-181b promotes hepatocarcinogenesis by targeting TIMP3. Oncogene 29(12):1787–1797. https://doi.org/10.1038/onc.2009.468
Wang Y, Yu Y, Tsuyada A, Ren X, Wu X, Stubblefield K et al (2011) Transforming growth factor-beta regulates the sphere-initiating stem cell-like feature in breast cancer through miRNA-181 and ATM. Oncogene 30(12):1470–1480. https://doi.org/10.1038/onc.2010.531
Wang B, Jin H, Shu B, Mira RR, Chen D (2015) Chondrocytes-specific expression of Osteoprotegerin modulates osteoclast formation in metaphyseal bone. Sci Rep 5:13667. https://doi.org/10.1038/srep13667
Wornham DP, Hajjawi MO, Orriss IR, Arnett TR (2014) Strontium potently inhibits mineralisation in bone-forming primary rat osteoblast cultures and reduces numbers of osteoclasts in mouse marrow cultures. Osteoporos Int 25(10):2477–2484. https://doi.org/10.1007/s00198-014-2791-5
Yamaguchi M, Weitzmann MN (2012) The intact strontium ranelate complex stimulates osteoblastogenesis and suppresses osteoclastogenesis by antagonizing NF-kappaB activation. Mol Cell Biochem 359(1–2):399–407. https://doi.org/10.1007/s11010-011-1034-8
Yang F, Yang D, Tu J, Zheng Q, Cai L, Wang L (2011) Strontium enhances osteogenic differentiation of mesenchymal stem cells and in vivo bone formation by activating Wnt/catenin signaling. Stem Cells 29(6):981–991. https://doi.org/10.1002/stem.646
Yao Q, Yu C, Zhang X, Zhang K, Guo J, Song L (2017) Wnt/beta-catenin signaling in osteoblasts regulates global energy metabolism. Bone 97:175–183. https://doi.org/10.1016/j.bone.2017.01.028
Zeng XZ, He LG, Wang S, Wang K, Zhang YY, Tao L et al (2016) Aconine inhibits RANKL-induced osteoclast differentiation in RAW264.7 cells by suppressing NF-kappaB and NFATc1 activation and DC-STAMP expression. Acta Pharmacol Sin 37(2):255–263. https://doi.org/10.1038/aps.2015.85
Zhao B, Grimes SN, Li S, Hu X, Ivashkiv LB (2012) TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibited by transcription factor RBP-J. J Exp Med 209(2):319–334. https://doi.org/10.1084/jem.20111566
Zhong ZDA, Zahatnansky J, Snider J, Van Wieren E, Diegel CR, Williams BO (2015) Wntless spatially regulates bone development through -catenin-dependent and independent mechanisms. Dev Dyn 244(10):1347–1355. https://doi.org/10.1002/dvdy.24316
Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (NSFC81371989 and NSFC81270967), Guangdong Science and Technology Department Project (2015A030313776, 2016A050503008), Shenzhen Municipal Science and Technology Innovation Committee Project (CXZZ20151015151249563, CXZZ20150401152251209, JCYJ20140416122812013 and JCYJ20150403101146318), General Research Fund of Research Grant Council of Hong Kong (RGC 715213 and RGC 17205714), Strategic Research Theme of Biomedical Engineering and Nanotechnology, Shenzhen Science and Technology Funding (JCYJ20160429185449249), Guangdong Scientific Plan (2014A030313743), and HK RGC (Ref No. T13-402/17-N).
Author information
Authors and Affiliations
Contributions
(i) Design: T. Sun, S. Peng, and Z. Li; (ii) Acquisition of the data: T. Sun, Z. Li and Y. Feng; (iii) Analysis and interpretation of the data: T. Sun, Z. Li, S. Peng and Y. Feng; (iv) Figures: T. Sun; (v) Writing - original draft: T. Sun; (vi) Writing - review & editing: T. Sun and Z. Cai; (vii) Funding, resources and supervision: WW. Lu, S. Peng, Y. Feng and F. Leung; (viii) Experimental supports: X. Zhong, Z. Ning, T. Hou and L. Xiong.
Corresponding authors
Ethics declarations
Conflict of interest
All authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Sun, T., Li, Z., Zhong, X. et al. Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p. J. Cell Commun. Signal. 13, 85–97 (2019). https://doi.org/10.1007/s12079-018-0478-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12079-018-0478-y