Download PDF
Int J Sports Med 2024; 45(11): 856-866
DOI: 10.1055/a-2323-9749
Orthopedics & Biomechanics

LINC00963 Represses Osteogenic Differentiation of hBMSCs via the miR-10b-5p/RAP2A/AKT Axis

Zhixin Wu 
1   Histology & Embryology Teaching and Researching Section, Xi’an Medical University, Xi’an, China
,
Mingjuan Shi
2   Public Health College, Xi’an Medical University, Xi’an, China
,
Xuan Zhao
1   Histology & Embryology Teaching and Researching Section, Xi’an Medical University, Xi’an, China
,
Guifu Wu
3   Department Endocrinology, Shaanxi Province People Hospital, Xi’an, China
,
Huiyuan Zheng
1   Histology & Embryology Teaching and Researching Section, Xi’an Medical University, Xi’an, China
,
Yuanyuan Cui
1   Histology & Embryology Teaching and Researching Section, Xi’an Medical University, Xi’an, China
,
Yu Shang
4   Clinical College, Xi’an Medical University, Xi’an, China
› Author Affiliations Funding Information Shaanxi Science and Technology Department Fund Project — 2010K13-01-03
› Further Information
Also available at
    Permissions and Reprints

Abstract

Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is important for human bone formation. Long non-coding RNAs (lncRNAs) are critical regulators in osteogenic differentiation. This study aimed to explore the function and mechanisms of long intergenic non-protein coding RNA 963 (LINC00963) in affecting osteogenesis. Cell differentiation was assessed by alkaline phosphatase (ALP) activity detection and ALP staining assay. Meanwhile, levels of osteogenic marker genes, including RUNX family transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN), were detected by RT-qPCR and western blot. Cell proliferation and apoptosis were measured using CCK-8 assay and flow cytometry analysis. RNA immunoprecipitation (RIP), RNA pull-down and luciferase reporter assays were used to investigate the interaction between genes. LINC00963 expression was down-regulated in hBMSCs treated with osteogenic induction. LINC00963 overexpression inhibited hBMSCs differentiation, proliferation, and elevated apoptosis. LINC00963 acted as a competing endogenous RNA (ceRNA) to interact with miR-10b-5p and thereby regulated the expression level of Ras-related protein Rap-2a (RAP2A). LINC00963 regulated RAP2A to inhibit the level of phosphorylated AKT (p-AKT). LINC00963 inhibited hBMSCs differentiation, proliferation, and elevated apoptosis via the miR-10b-5p/RAP2A/AKT signaling, which might help improve the treatment of osteoporosis.

Keywords

postmenopausal osteoporosis - LINC00963 - differentiation - AKT

Supplementary Material



Publication History

Received: 22 August 2022

Accepted: 26 April 2024

Article published online:
28 July 2024

© 2024. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Armas LA, Recker RR. Pathophysiology of osteoporosis: new mechanistic insights. Endocrinol Metab Clin North Am 2012; 41: 475-486
    CrossrefPubMedSearch in Google Scholar
  • 2 Jackson RD, Mysiw WJ. Insights into the epidemiology of postmenopausal osteoporosis: the Women's Health Initiative. Semin Reprod Med 2014; 32: 454-462
    Thieme ConnectPubMedSearch in Google Scholar
  • 3 Vilaca T, Eastell R, Schini M. Osteoporosis in men. Lancet Diabetes Endocrinol 2022; 10: 273-283
    CrossrefPubMedSearch in Google Scholar
  • 4 Johnell O, Kanis JA. An estimate of the worldwide prevalence and disability associated with osteoporotic fractures. Osteoporos Int 2006; 17: 1726-1733
    CrossrefPubMedSearch in Google Scholar
  • 5 Shepstone L, Lenaghan E, Cooper C. et al. Screening in the community to reduce fractures in older women (SCOOP): a randomised controlled trial. Lancet 2018; 391: 741-747
    CrossrefPubMedSearch in Google Scholar
  • 6 De Souza MJ, Koltun KJ, Williams NI. The Role of Energy Availability in Reproductive Function in the Female Athlete Triad and Extension of its Effects to Men: An Initial Working Model of a Similar Syndrome in Male Athletes. Sports Med 2019; 49: 125-137
    CrossrefPubMedSearch in Google Scholar
  • 7 Nattiv A, De Souza MJ, Koltun KJ. et al. The Male Athlete Triad–A Consensus Statement From the Female and Male Athlete Triad Coalition Part 1: Definition and Scientific Basis. Clin J Sport Med 2021; 31: 345-353
    CrossrefPubMedSearch in Google Scholar
  • 8 Tenforde AS, Carlson JL, Sainani KL. et al. Sport and triad risk factors influence bone mineral density in collegiate athletes. Med Sci Sports Exerc 2018; 50: 2536-2543
    CrossrefPubMedSearch in Google Scholar
  • 9 Scofield KL, Hecht S. Bone health in endurance athletes: runners, cyclists, and swimmers. Curr Sports Med Rep 2012; 11: 328-334
    CrossrefPubMedSearch in Google Scholar
  • 10 Sale C, Elliott-Sale KJ. Nutrition and athlete bone health. Sports Med 2019; 49: 139-151
    CrossrefPubMedSearch in Google Scholar
  • 11 Samsonraj RM, Paradise CR, Dudakovic A. et al. Validation of osteogenic properties of cytochalasin D by high-resolution RNA-sequencing in mesenchymal stem cells derived from bone marrow and adipose tissues. Stem Cells Dev 2018; 27: 1136-1145
    CrossrefPubMedSearch in Google Scholar
  • 12 Huang Y, Yin Y, Gu Y. et al. Characterization and immunogenicity of bone marrow-derived mesenchymal stem cells under osteoporotic conditions. Sci China Life Sci 2020; 63: 429-442
    CrossrefPubMedSearch in Google Scholar
  • 13 Huang S, Zhu X, Xiao D. et al. LncRNA SNHG1 was down-regulated after menopause and participates in postmenopausal osteoporosis. Biosci Rep 2019; 39
    CrossrefPubMedSearch in Google Scholar
  • 14 Xiaoling G, Shuaibin L, Kailu L. MicroRNA-19b-3p promotes cell proliferation and osteogenic differentiation of BMSCs by interacting with lncRNA H19. BMC Med Genet 2020; 21: 11
    CrossrefPubMedSearch in Google Scholar
  • 15 Jiao H, Jiang S, Wang H. et al. Upregulation of LINC00963 facilitates melanoma progression through miR-608/NACC1 pathway and predicts poor prognosis. Biochem Biophys Res Commun 2018; 504: 34-39
    CrossrefPubMedSearch in Google Scholar
  • 16 Xie LB, Chen B, Liao X. et al. LINC00963 targeting miR-128-3p promotes acute kidney injury process by activating JAK2/STAT1 pathway. J Cell Mol Med 2020; 24: 5555-5564
    CrossrefPubMedSearch in Google Scholar
  • 17 Fei Q, Bai X, Lin J. et al. Identification of aberrantly expressed long non-coding RNAs in postmenopausal osteoporosis. Int J Mol Med 2018; 41: 3537-3550
    CrossrefPubMedSearch in Google Scholar
  • 18 Ren L, Guo L, Kou N. et al. LncRNA LINC00963 promotes osteogenic differentiation of hBMSCs and alleviates osteoporosis progression by targeting miRNA-760/ETS1 axis. Autoimmunity 2021; 54: 313-325
    CrossrefPubMedSearch in Google Scholar
  • 19 Zhang H, Bian C, Tu S. et al. Integrated analysis of lncRNA-miRNA-mRNA ceRNA network in human aortic dissection. BMC Genomics 2021; 22: 724
    CrossrefPubMedSearch in Google Scholar
  • 20 Mahesh G, Biswas R. MicroRNA-155: A master regulator of inflammation. J Interferon Cytokine Res 2019; 39: 321-330
    CrossrefPubMedSearch in Google Scholar
  • 21 Li Z, Zhang W, Huang Y. MiRNA-133a is involved in the regulation of postmenopausal osteoporosis through promoting osteoclast differentiation. Acta Biochim Biophys Sin (Shanghai) 2018; 50: 273-280
    CrossrefPubMedSearch in Google Scholar
  • 22 Fu Y, Xu Y, Chen S. et al. MiR-151a-3p promotes postmenopausal osteoporosis by targeting SOCS5 and activating JAK2/STAT3 signaling. Rejuvenation Res 2020; 23: 313-323
    CrossrefPubMedSearch in Google Scholar
  • 23 Singh R, Ha SE, Wei L. et al. miR-10b-5p rescues diabetes and gastrointestinal dysmotility. Gastroenterology 2021; 160: 1662-1678.e1618
    CrossrefPubMedSearch in Google Scholar
  • 24 Wu L, Chen Y, Chen Y. et al. Effect of HIF-1α/miR-10b-5p/PTEN on hypoxia-induced cardiomyocyte apoptosis. J Am Heart Assoc 2019; 8: e011948
    CrossrefPubMedSearch in Google Scholar
  • 25 Quan R, Liang W, Li H. et al. Silencing of miR-10b-5p alleviates the mechanical stretch-induced proliferation of HASMCs. Tissue Cell 2022; 74: 101700
    CrossrefPubMedSearch in Google Scholar
  • 26 Wang M, Li J, Ye Y. et al. SHED-derived conditioned exosomes enhance the osteogenic differentiation of PDLSCs via Wnt and BMP signaling in vitro. Differentiation 2020; 111: 1-11
    CrossrefPubMedSearch in Google Scholar
  • 27 Carlevaro-Fita J, Johnson R. Global positioning system: Understanding long noncoding RNAs through subcellular localization. Mol Cell 2019; 73: 869-883
    CrossrefPubMedSearch in Google Scholar
  • 28 Turchinovich A, Weiz L, Langheinz A. et al. Characterization of extracellular circulating microRNA. Nucleic Acids Res 2011; 39: 7223-7233
    CrossrefPubMedSearch in Google Scholar
  • 29 Wang L, Zhan W, Xie S. et al. Over-expression of Rap2a inhibits glioma migration and invasion by down-regulating p-AKT. Cell Biol Int 2014; 38: 326-334
    CrossrefPubMedSearch in Google Scholar
  • 30 Sun Y, Wang TE, Hu Q. et al. Systematic comparation of the biological and transcriptomic landscapes of human amniotic mesenchymal stem cells under serum-containing and serum-free conditions. Stem Cell Res Ther 2022; 13: 490
    CrossrefPubMedSearch in Google Scholar
  • 31 Ko NY, Chen LR, Chen KH. The role of micro RNA and long-non-coding RNA in osteoporosis. Int J Mol Sci 2020; 21
    CrossrefPubMedSearch in Google Scholar
  • 32 Yuan Y, Guo J, Zhang L. et al. MiR-214 Attenuates the osteogenic effects of mechanical loading on osteoblasts. Int J Sports Med 2019; 40: 931-940
    Thieme ConnectPubMedSearch in Google Scholar
  • 33 Zhang N, Zeng X, Sun C. et al. LncRNA LINC00963 promotes tumorigenesis and radioresistance in breast cancer by sponging miR-324-3p and inducing ACK1 expression. Mol Ther Nucleic Acids 2019; 18: 871-881
    CrossrefPubMedSearch in Google Scholar
  • 34 Wu Z, Wang W, Wang Y. et al. Long noncoding RNA LINC00963 promotes breast cancer progression by functioning as a molecular sponge for microRNA-625 and thereby upregulating HMGA1. Cell Cycle 2020; 19: 610-624
    CrossrefPubMedSearch in Google Scholar
  • 35 Ge G, Yang D, Tan Y. et al. miR-10b-5p regulates C2C12 myoblasts proliferation and differentiation. Biosci Biotechnol Biochem 2019; 83: 291-299
    CrossrefPubMedSearch in Google Scholar
  • 36 Chen R, Liao X, Chen F. et al. Circulating microRNAs, miR-10b-5p, miR-328-3p, miR-100 and let-7, are associated with osteoblast differentiation in osteoporosis. Int J Clin Exp Pathol 2018; 11: 1383-1390
    PubMedSearch in Google Scholar
  • 37 Li Y, Chen D, Li Y. et al. Oncogenic cAMP responsive element binding protein 1 is overexpressed upon loss of tumor suppressive miR-10b-5p and miR-363-3p in renal cancer. Oncol Rep 2016; 35: 1967-1978
    CrossrefPubMedSearch in Google Scholar
  • 38 Zheng X, Zhao W, Ji P. et al. High expression of Rap2A is associated with poor prognosis of patients with hepatocellular carcinoma. Int J Clin Exp Pathol 2017; 10: 9607-9613
    PubMedSearch in Google Scholar
  • 39 Pompura SL, Dominguez-Villar M. The PI3K/AKT signaling pathway in regulatory T-cell development, stability, and function. J Leukoc Biol 2018;
    CrossrefPubMedSearch in Google Scholar
  • 40 Revathidevi S, Munirajan AK. Akt in cancer: Mediator and more. Semin Cancer Biol 2019; 59: 80-91
    CrossrefPubMedSearch in Google Scholar
  • 41 Wu JX, Du WQ, Wang XC. et al. Rap2a serves as a potential prognostic indicator of renal cell carcinoma and promotes its migration and invasion through up-regulating p-Akt. Sci Rep 2017; 7: 6623
    CrossrefPubMedSearch in Google Scholar
  • 42 Wu JH, Tian XY, An QM. et al. LINC00963 promotes hepatocellular carcinoma progression by activating PI3K/AKT pathway. Eur Rev Med Pharmacol Sci 2018; 22: 1645-1652
    CrossrefPubMedSearch in Google Scholar