Elevation of MiR-302b Prevents Multiple Myeloma Cell Growth and Bone Destruction by Blocking DKK1 Secretion

Overview

Journal Cancer Cell Int

Publisher Biomed Central

Date 2021 Apr 1

PMID 33789678

Citations 5

Authors

Zheyu Wu

Yufeng Zhang

Zhiqiang Yang

Yufan Zhu

Yuanlong Xie

Fuling Zhou

Lin Cai

Affiliations

Soon will be listed here.

Abstract

Background: Myeloma bone disease (MBD) is a severe complication of multiple myeloma (MM) mainly due to an imbalance between enhanced osteoclast activity and reduced osteoblast function. Previous studies have demonstrated that miRNAs play a vital role in the osteogenic differentiation of mesenchymal stromal cells (MSCs) in MM. However, the value of miR‑302b in MBD remains to be further elucidated. The aim of this study is to explore the role of miR‑302b in the regulation of MBD osteogenic differentiation and evaluate the potential of a new therapeutic strategy for the clinical treatment of MBD.

Method: Our previous research demonstrated that MiR-302b belongs to the miR-302 cluster and is able to inhibit tumor growth and osteolysis in an orthotopic osteosarcoma xenograft tumor mouse model. In this study, we first transfected miR-302b mimics, miR-302b inhibitor, and miR-302b NC into MM1.S and RPMI8226 MM cells to detect the correlation between miR-302b expression in the pathological specimens and the clinicopathological features by qPCR, the target correlation between miR-302b and DKK1 by immunohistochemistry, qPCR and Western blot, and the correlation between miR-302b and the Wnt/β-catenin signaling pathway by Western blot. The effect of miR-302b on osteoblastogenesis was also studied in a subperiosteal tumorigenesis model of NOD/SCID nude mice.

Results: We found that increased miR-302b suppressed cell proliferation and induced cell apoptosis in RPMI 8226 and MM1.S cells. TargetScan online bioinformatic analysis predicted that miR-302b is able to bind to 3'UTR of DKK1 mRNA. Target binding of miR-302b to DKK1 was demonstrated by dual-luciferase reporter assay, qPCR, Western blot and immunohistochemistry, indicating that miR-302b is able to degrade DKK1 in RPMI 8226 and MM1.S cells. The model of co-culturing MM cells with preosteoblast MC3T3-E1 cells showed that miR-302b inhibits MM-induced suppression of osteoblast differentiation. Western blotting showed that miR-302b promotes the Wnt/β-catenin signaling pathway in MM cells. Micro-CT and immunohistochemistry results showed that miR-302b suppresses myeloma bone destruction in vivo.

Conclusion: miR-302b is able to target DKK1 and promote the Wnt/β-catenin signaling pathway in MM.

Citing Articles

Advances in the pathogenesis of multiple myeloma bone disease.

Hu C, Kuang C, Zhou W Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2023; 48(9):1403-1410.

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Drug Discovery of DKK1 Inhibitors.

Jiang H, Zhang Z, Yu Y, Chu H, Yu S, Yao S Front Pharmacol. 2022; 13:847387.

PMID: 35355709 PMC: 8959454. DOI: 10.3389/fphar.2022.847387.

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Perspectives on miRNAs Targeting DKK1 for Developing Hair Regeneration Therapy.

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References

Anderson K . Progress and Paradigms in Multiple Myeloma. Clin Cancer Res. 2017; 22(22):5419-5427. PMC: 5300651. DOI: 10.1158/1078-0432.CCR-16-0625. View

Misso G, Zappavigna S, Castellano M, De Rosa G, Di Martino M, Tagliaferri P . Emerging pathways as individualized therapeutic target of multiple myeloma. Expert Opin Biol Ther. 2013; 13 Suppl 1:S95-109. DOI: 10.1517/14712598.2013.807338. View

Terpos E, Dimopoulos M . Myeloma bone disease: pathophysiology and management. Ann Oncol. 2005; 16(8):1223-31. DOI: 10.1093/annonc/mdi235. View

Delgado-Calle J, Anderson J, Cregor M, Condon K, Kuhstoss S, Plotkin L . Genetic deletion of Sost or pharmacological inhibition of sclerostin prevent multiple myeloma-induced bone disease without affecting tumor growth. Leukemia. 2017; 31(12):2686-2694. PMC: 5699973. DOI: 10.1038/leu.2017.152. View

Xie Y, Sun W, Deng Z, Zhu X, Hu C, Cai L . MiR-302b Suppresses Osteosarcoma Cell Migration and Invasion by Targeting Runx2. Sci Rep. 2017; 7(1):13388. PMC: 5645461. DOI: 10.1038/s41598-017-13353-9. View

Raje N, Roodman G . Advances in the biology and treatment of bone disease in multiple myeloma. Clin Cancer Res. 2011; 17(6):1278-86. DOI: 10.1158/1078-0432.CCR-10-1804. View

Rossi M, Di Martino M, Morelli E, Leotta M, Rizzo A, Grimaldi A . Molecular targets for the treatment of multiple myeloma. Curr Cancer Drug Targets. 2012; 12(7):757-67. DOI: 10.2174/156800912802429300. View

Negaard H, Iversen N, Bowitz-Lothe I, Sandset P, Steinsvik B, Ostenstad B . Increased bone marrow microvascular density in haematological malignancies is associated with differential regulation of angiogenic factors. Leukemia. 2008; 23(1):162-9. DOI: 10.1038/leu.2008.255. View

Garayoa M, Garcia J, Santamaria C, Garcia-Gomez A, Blanco J, Pandiella A . Mesenchymal stem cells from multiple myeloma patients display distinct genomic profile as compared with those from normal donors. Leukemia. 2009; 23(8):1515-27. DOI: 10.1038/leu.2009.65. View

10.

Morgan G, Walker B, Davies F . The genetic architecture of multiple myeloma. Nat Rev Cancer. 2012; 12(5):335-48. DOI: 10.1038/nrc3257. View

11.

Yadav S, Pandey A, Shukla A, Talwelkar S, Kumar A, Pant A . miR-497 and miR-302b regulate ethanol-induced neuronal cell death through BCL2 protein and cyclin D2. J Biol Chem. 2011; 286(43):37347-57. PMC: 3199482. DOI: 10.1074/jbc.M111.235531. View

12.

Manier S, Salem K, Park J, Landau D, Getz G, Ghobrial I . Genomic complexity of multiple myeloma and its clinical implications. Nat Rev Clin Oncol. 2016; 14(2):100-113. DOI: 10.1038/nrclinonc.2016.122. View

13.

Tardif G, Hum D, Pelletier J, Duval N, Martel-Pelletier J . Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes. BMC Musculoskelet Disord. 2009; 10:148. PMC: 2792220. DOI: 10.1186/1471-2474-10-148. View

14.

Umezu T, Tadokoro H, Azuma K, Yoshizawa S, Ohyashiki K, Ohyashiki J . Exosomal miR-135b shed from hypoxic multiple myeloma cells enhances angiogenesis by targeting factor-inhibiting HIF-1. Blood. 2014; 124(25):3748-57. PMC: 4263983. DOI: 10.1182/blood-2014-05-576116. View

15.

Amodio N, Di Martino M, Foresta U, Leone E, Lionetti M, Leotta M . miR-29b sensitizes multiple myeloma cells to bortezomib-induced apoptosis through the activation of a feedback loop with the transcription factor Sp1. Cell Death Dis. 2012; 3:e436. PMC: 3542610. DOI: 10.1038/cddis.2012.175. View

16.

Di Martino M, Leone E, Amodio N, Foresta U, Lionetti M, Pitari M . Synthetic miR-34a mimics as a novel therapeutic agent for multiple myeloma: in vitro and in vivo evidence. Clin Cancer Res. 2012; 18(22):6260-70. PMC: 4453928. DOI: 10.1158/1078-0432.CCR-12-1708. View

17.

Khalife J, Ghose J, Martella M, Viola D, Rocci A, Troadec E . MiR-16 regulates crosstalk in NF-κB tolerogenic inflammatory signaling between myeloma cells and bone marrow macrophages. JCI Insight. 2019; 4(21). PMC: 6948777. DOI: 10.1172/jci.insight.129348. View

18.

Li J, Yu J, Zhang H, Wang B, Guo H, Bai J . Exosomes-Derived MiR-302b Suppresses Lung Cancer Cell Proliferation and Migration via TGFβRII Inhibition. Cell Physiol Biochem. 2016; 38(5):1715-26. DOI: 10.1159/000443111. View

19.

Zhang Y, Hu H, Song L, Cai L, Wei R, Jin W . Epirubicin-mediated expression of miR-302b is involved in osteosarcoma apoptosis and cell cycle regulation. Toxicol Lett. 2013; 222(1):1-9. DOI: 10.1016/j.toxlet.2013.06.242. View

20.

Vyzoukaki R, Tsirakis G, Pappa C, Devetzoglou M, Tzardi M, Alexandrakis M . The Impact of Mast Cell Density on the Progression of Bone Disease in Multiple Myeloma Patients. Int Arch Allergy Immunol. 2016; 168(4):263-8. DOI: 10.1159/000443275. View