miR-363-3p is activated by MYB and regulates osteoporosis pathogenesis via PTEN/PI3K/AKT signaling pathway

  • Mingyi Li
  • Ruolan LuoEmail author
  • Wenjian Yang
  • Zhen Zhou
  • Chenxia Li


Osteoporosis results from the imbalance between osteogenesis and bone resorption mediated by osteoblasts and osteoclasts. During the disease process of osteoporosis, the alteration of gene expression occurs, which lead to the disease progression. MicroRNAs (miRNAs) have been previously demonstrated to be modulators for bone metabolism via regulation of osteoblast and osteoclast differentiation. In the present study, we detected the expression levels of five osteoporosis-related miRNAs in bone and serum samples of patient with or without osteoporosis. The downstream molecular mechanism of miR-363-3p was analyzed and detected by using bioinformatics analysis and mechanism experiment. The upstream transcription factor of miR-363-3p was analyzed by applying bioinformatics analysis and ChIP assay and luciferase reporter assay. The role of this pathway in osteoclastogenesis was demonstrated by functional assays. MiR-363-3p was significantly highly expressed in osteoporotic samples. Mechanistically, miR-363-3p promotes osteoclastogenesis and inhibits osteogenic differentiation by targeting PTEN and therefore activating PI3K/AKT signaling pathway. MiR-363-3p was activated by its upstream transcription activator MYB. This study revealed that MYB-induced upregulation of miR-363-3p regulates osteoporosis pathogenesis via PTEN/PI3K/AKT signaling pathway.


MYB miR-363-3p Osteoporosis PTEN PI3K/AKT signaling pathway 



The authors thank all members contributed to this study.

Compliance with ethical standards

This study was approved by the Research Ethics Committee of Xiangyang No.1 People’s Hospital.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11626_2019_344_Fig7_ESM.png (125 kb)
Supplementary figure 1

The expression of several miRNAs in the bone and serum samples. A-B. The level of indicated miRNAs (miR-214, miR-34a, miR-103a, miR-503 and miR-148a), whose function in osteoporosis has already been reported previously, was detected using qRT-PCR analysis in 6 pairs of bone tissues (A) and 6 pairs of serum samples (B). *P < 0.05, **P < 0.01. (PNG 124 kb)

11626_2019_344_MOESM1_ESM.tif (4.8 mb)
High Resolution Image (TIF 4913 kb)
11626_2019_344_Fig8_ESM.png (459 kb)
Supplementary figure 2

The expression of genes involved in miR-363-3p/PTEN/PI3K/AKT axis during osteoclast differentiation. A-B. qRT-PCR analysis indicated that miR-363-3p level was increased while PTEN expression decreased in a time-dependent manner in CD14 + PBMCs under M-CSF and RANKL treatment. C. The protein levels of PTEN, AKT and p-AKT in CD14 + PBMCs when co-treating with M-CSF and RANKL for 0, 24, 48, and 72 h were evaluated by Western blot analysis. *P < 0.05, **P < 0.01. (PNG 459 kb)

11626_2019_344_MOESM2_ESM.tif (8.9 mb)
High Resolution Image (TIF 9127 kb)
11626_2019_344_MOESM3_ESM.doc (36 kb)
Supplementary table 1 The expression of genes in the bone tissues from osteoporotic and non-osteoporotic patients. Low/high by the sample median. Paired-sample t test. P < 0.05 was thought to be of statistical significance. (DOC 35 kb)
11626_2019_344_MOESM4_ESM.docx (15 kb)
Supplementary table 2 The expression of genes in the serum samples from osteoporotic and non-osteoporotic patients. Low/high by the sample median. Paired-sample t test. P < 0.05 was thought to be of statistical significance. (DOCX 14 kb)


  1. Adler RA (2016) Osteoporosis treatment: complexities and challenges. J Endocrinol Investig 39:719–720CrossRefGoogle Scholar
  2. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297CrossRefGoogle Scholar
  3. Bennett CN, Longo KA, Wright WS, Suva LJ, Lane TF, Hankenson KD, MacDougald OA (2005) Regulation of osteoblastogenesis and bone mass by Wnt10b. Proc Natl Acad Sci U S A 102:3324–3329CrossRefGoogle Scholar
  4. Bhagirath D, Yang TL, Bucay N, Sekhon K, Majid S, Shahryari V, Dahiya R, Tanaka Y, Saini S (2018) MicroRNA-1246 is an exosomal biomarker for aggressive prostate cancer. Cancer Res 78:1833–1844. CrossRefGoogle Scholar
  5. Bollerslev J, Wilson SG, Dick IM, Islam FM, Ueland T, Palmer L, Devine A, Prince RL (2005) LRP5 gene polymorphisms predict bone mass and incident fractures in elderly Australian women. Bone 36:599–606CrossRefGoogle Scholar
  6. Chen C, Cheng P, Xie H, Zhou HD, Wu XP, Liao EY, Luo XH (2014) MiR-503 regulates osteoclastogenesis via targeting RANK. J Bone Miner Res 29:338–347CrossRefGoogle Scholar
  7. Feng Y, Zhou S, Li G, Hu C, Zou W, Zhang H, Sun L (2016) Nuclear factor-kappaB-dependent microRNA-130a upregulation promotes cervical cancer cell growth by targeting phosphatase and tensin homolog. Arch Biochem Biophys 598:57–65CrossRefGoogle Scholar
  8. Fiore R, Khudayberdiev S, Christensen M, Siegel G, Flavell SW, Kim TK, Greenberg ME, Schratt G (2009) Mef2-mediated transcription of the miR379-410 cluster regulates activity-dependent dendritogenesis by fine-tuning Pumilio2 protein levels. EMBO J 28:697–710CrossRefGoogle Scholar
  9. Formosa A, Markert EK, Lena AM, Italiano D, Finazzi-Agro E, Levine AJ, Bernardini S, Garabadgiu AV, Melino G, Candi E (2014) MicroRNAs, miR-154, miR-299-5p, miR-376a, miR-376c, miR-377, miR-381, miR-487b, miR-485-3p, miR-495 and miR-654-3p, mapped to the 14q32.31 locus, regulate proliferation, apoptosis, migration and invasion in metastatic prostate cancer cells. Oncogene 33:5173–5182CrossRefGoogle Scholar
  10. He L, Thomson JM, Hemann MT, Hernando-Monge E, Mu D, Goodson S, Powers S, Cordon-Cardo C, Lowe SW, Hannon GJ, Hammond SM (2005) A microRNA polycistron as a potential human oncogene. Nature 435:828–833CrossRefGoogle Scholar
  11. He XX, Chang Y, Meng FY, Wang MY, Xie QH, Tang F, Li PY, Song YH, Lin JS (2012) MicroRNA-375 targets AEG-1 in hepatocellular carcinoma and suppresses liver cancer cell growth in vitro and in vivo. Oncogene 31:3357–3369CrossRefGoogle Scholar
  12. Jia J, Tian Q, Ling S, Liu Y, Yang S, Shao Z (2013) miR-145 suppresses osteogenic differentiation by targeting Sp7. FEBS Lett 587:3027–3031CrossRefGoogle Scholar
  13. Li WF, Hou SX, Yu B, Li MM, Ferec C, Chen JM (2010) Genetics of osteoporosis: accelerating pace in gene identification and validation. Hum Genet 127:249–285CrossRefGoogle Scholar
  14. Li KC, Chang YH, Yeh CL, Hu YC (2015a) Healing of osteoporotic bone defects by baculovirus-engineered bone marrow-derived MSCs expressing MicroRNA sponges. Biomaterials 74:155–166CrossRefGoogle Scholar
  15. Li H, Li T, Fan J, Li T, Fan L, Wang S, Weng X, Han Q, Zhao RC (2015b) miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway. Cell Death Differ 22:1935–1945. CrossRefGoogle Scholar
  16. Lin CH, Jackson AL, Guo J, Linsley PS, Eisenman RN (2009) Myc-regulated microRNAs attenuate embryonic stem cell differentiation. EMBO J 28:3157–3170CrossRefGoogle Scholar
  17. Mulari MT, Qu Q, Harkonen PL, Vaananen HK (2004) Osteoblast-like cells complete osteoclastic bone resorption and form new mineralized bone matrix in vitro. Calcif Tissue Int 75:253–261CrossRefGoogle Scholar
  18. Oralova V, Matalova E, Killinger M, Knopfova L, Smarda J, Buchtova M (2017) Osteogenic potential of the transcription factor c-MYB. Calcif Tissue Int 100:311–322CrossRefGoogle Scholar
  19. Pal MK, Jaiswar SP, Dwivedi VN, Tripathi AK, Dwivedi A, Sankhwar P (2015) MicroRNA: a new and promising potential biomarker for diagnosis and prognosis of ovarian cancer. Cancer biology & medicine 12:328–341Google Scholar
  20. Rana TM (2007) Illuminating the silence: understanding the structure and function of small RNAs. Nat Rev Mol Cell Biol 8:23–36CrossRefGoogle Scholar
  21. Rouse R, Rosenzweig B, Shea K, Knapton A, Stewart S, Xu L, Chockalingam A, Zadrozny L, Thompson K (2017) MicroRNA biomarkers of pancreatic injury in a canine model. Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie 69:33–43CrossRefGoogle Scholar
  22. Sandbothe M, Buurman R, Reich N, Greiwe L, Vajen B, Gürlevik E, Schäffer V, Eilers M, Kühnel F, Vaquero A, Longerich T, Roessler S, Schirmacher P, Manns MP, Illig T, Schlegelberger B, Skawran B (2017) The microRNA-449 family inhibits TGF-β-mediated liver cancer cell migration by targeting SOX4. J Hepatol 66:1012–1021. CrossRefGoogle Scholar
  23. Shi K, Lu J, Zhao Y, Wang L, Li J, Qi B, Li H, Ma C (2013) MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix. Bone 55:487–494CrossRefGoogle Scholar
  24. Sorensen MG, Henriksen K, Schaller S, Henriksen DB, Nielsen FC, Dziegiel MH, Karsdal MA (2007) Characterization of osteoclasts derived from CD14+ monocytes isolated from peripheral blood. J Bone Miner Metab 25:36–45CrossRefGoogle Scholar
  25. Sun R, Li C, Zhang J, Li F, Ma L, Tan Y, Wang Q, Zhang B (2017) Differential expression of microRNAs during fiber development between fuzzless-lintless mutant and its wild-type allotetraploid cotton. Sci Rep 7:3CrossRefGoogle Scholar
  26. Wang X, Guo B, Li Q, Peng J, Yang Z, Wang A, Li D, Hou Z, Lv K, Kan G, Cao H, Wu H, Song J, Pan X, Sun Q, Ling S, Li Y, Zhu M, Zhang P, Peng S, Xie X, Tang T, Hong A, Bian Z, Bai Y, Lu A, Li Y, He F, Zhang G, Li Y (2013) miR-214 targets ATF4 to inhibit bone formation. Nat Med 19:93–100CrossRefGoogle Scholar
  27. Wang O, Hu Y, Gong S, Xue Q, Deng Z, Wang L, Liu H, Tang H, Guo X, Chen J, Jia X, Xu Y, Lan L, Lei C, Dong H, Yuan G, Fu Q, Wei Y, Xia W, Xu L (2015) A survey of outcomes and management of patients post fragility fractures in China. Osteoporosis international : a journal established as result of cooperation between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA 26:2631–2640CrossRefGoogle Scholar
  28. Wang W, Oguz G, Lee PL, Bao Y, Wang P, Terp MG, Ditzel HJ, Yu Q (2018a) KDM4B-regulated unfolded protein response as a therapeutic vulnerability in PTEN-deficient breast cancer. J Exp Med 215:2833–2849. CrossRefGoogle Scholar
  29. Wang Y, Zhang CY, Xia RH, Han J, Sun B, Sun SY, Li J (2018b) The MYB/miR-130a/NDRG2 axis modulates tumor proliferation and metastatic potential in salivary adenoid cystic carcinoma. Cell Death Dis 9:917CrossRefGoogle Scholar
  30. Xiao J, Lv D, Zhou J, Bei Y, Chen T, Hu M, Zhou Q, Fu S, Huang Q (2017) Therapeutic inhibition of miR-4260 suppresses colorectal cancer via targeting MCC and SMAD4. Theranostics 7:1901–1913CrossRefGoogle Scholar
  31. Xie R, Zhang J, Ma Y, Pan X, Dong C, Pang S, He S, Deng L, Yi S, Zheng Y, Lv Q (2017) Combined analysis of mRNA and miRNA identifies dehydration and salinity responsive key molecular players in citrus roots. Sci Rep 7:42094CrossRefGoogle Scholar
  32. Yang CH, Yue J, Fan M, Pfeffer LM (2010) IFN induces miR-21 through a signal transducer and activator of transcription 3-dependent pathway as a suppressive negative feedback on IFN-induced apoptosis. Cancer Res 70:8108–8116CrossRefGoogle Scholar
  33. Zhao W, Dong Y, Wu C, Ma Y, Jin Y, Ji Y (2015a) MiR-21 overexpression improves osteoporosis by targeting RECK. Mol Cell Biochem 405:125–133CrossRefGoogle Scholar
  34. Zhao C, Sun W, Zhang P, Ling S, Li Y, Zhao D, Peng J, Wang A, Li Q, Song J, Wang C, Xu X, Xu Z, Zhong G, Han B, Chang YZ, Li Y (2015b) miR-214 promotes osteoclastogenesis by targeting Pten/PI3k/Akt pathway. RNA Biol 12:343–353CrossRefGoogle Scholar
  35. Zhou R, Hu G, Gong AY, Chen XM (2010) Binding of NF-kappaB p65 subunit to the promoter elements is involved in LPS-induced transactivation of miRNA genes in human biliary epithelial cells. Nucleic Acids Res 38:3222–3232CrossRefGoogle Scholar
  36. Zuo B, Zhu JF, Li J, Wang CD, Zhao XY, Cai GQ, Li Z, Peng J, Wang P, Shen C, Huang Y, Xu J, Zhang XL, Chen XD (2014) microRNA-103a functions as a mechno-sensitive microRNA to inhibit bone formation through targeting Runx2. J Bone Miner Res 30:330–345. CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Endocrine Department, Xiangyang No.1 People’s HospitalHubei University of MedicineXiangyangChina
  2. 2.Endocrine DepartmentXiangyang Hospital of Traditional Chinese MedicineXiangyangChina

Personalised recommendations