Advances in Clinical and Experimental Medicine
2019, vol. 28, nr 10, October, p. 1293–1300
Publication type: original article
miR-29a promotes osteoblast proliferation by downregulating DKK-1 expression and activating Wnt/β-catenin signaling pathway
1 First Hospital Of Anhui Medical University, Hefei, China
Background. MicroRNA (miRNA) is a kind of non-coding small RNA with a negative regulating function. Some miRNAs play a role in regulating the differentiation and function of osteoblasts, chondrocytes and osteoclasts.
Objectives. In this study, we analyzed the role of miR-29a and dickkopf-1 (DKK-1) in osteoblast differentiation.
Material and Methods. Specimens were collected from the surgical resection of pathological ankylosing spondylitis (AS) tissue and some normal tissues. The expression of miR-29a, DKK-1 and β-catenin in normal and AS tissues were detected with real-time polymerase chain reaction (RT-PCR) and western blotting. Cell proliferation was detected with a Cell Counting Kit-8, cell migration and invasion were determined using a Transwell system and cell apoptosis was analyzed with flow cytometry. The luciferase reporter gene plasmid pGL3-DKK-1 and a point-mutation of the luciferase reporter gene plasmid mut-pGL3-DKK-1 were constructed.
Results. It was found that miR-29a could promote the proliferation of hFOB1.19 cells, while DKK-1 inhibited their proliferation. Also, miR-29a was able to inhibit the apoptosis of hFOB1.19 cells, while DKK-1 was able to promote the apoptosis of hFOB1.19 cells. When it comes to the invasion and migration of hFOB1.19 cells, miR-29a was found to promote it, while DKK-1 did not.
Conclusion. These findings will lead to a better understanding of the proliferation and differentiation of osteoblasts and will provide new insights for the treatment of this disease.
β-catenin, Dkk-1, miR-29a, si-RNA
- Huang W, Yang S, Shao J, Li YP. Signaling and transcriptional regulation in osteoblast commitment and differentiation. Front Biosci. 2007;12:3068–3092.
- Liu DQ, Zhang J, Song HN, Zheng J, Wang XX. Expression of Smad4 and Smad7 of BMP signaling pathway in oral squamous cell carcinoma [in Chinese]. Shanghai Kou Qiang Yi Xue. 2013;22:492–497.
- Chen L, Holmstrom K, Qiu W, et al. MicroRNA-34a inhibits osteoblast differentiation and in vivo bone formation of human stromal stem cells. Stem Cells. 2014;32(4):902–912.
- Liu X, Xu H, Kou J, Wang Q, Zheng X, Yu T. MiR-9 promotes osteoblast differentiation of mesenchymal stem cells by inhibiting DKK1 gene expression. Mol Biol Rep. 2016;43(9):939–946.
- Wang FS, Chuang PC, Lin CL, et al. MicroRNA-29a protects against glucocorticoid-induced bone loss and fragility in rats by orchestrating bone acquisition and resorption. Arthritis Rheum. 2013;65(6):1530–1540.
- Ko JY, Chuang PC, Chen MW, et al. MicroRNA-29a ameliorates glucocorticoid-induced suppression of osteoblast differentiation by regulating beta-catenin acetylation. Bone. 2013;57(2):468–475.
- Li Z, Hassan MQ, Jafferji M, et al. Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation. J Biol Chem. 2009;284(23):15676–15684.
- Kapinas K, Kessler C, Ricks T, Gronowicz G, Delany AM. MiR-29 modulates Wnt signaling in human osteoblasts through a positive feedback loop. J Biol Chem. 2010;285(33):25221–25231.
- Sudo R, Sato F, Azechi T, Wachi H. MiR-29-mediated elastin downregulation contributes to inorganic phosphorus-induced osteoblastic differentiation in vascular smooth muscle cells. Genes Cells. 2015;20(12):1077–1087.
- Tan K, Peng YT, Guo P. MiR-29a promotes osteogenic differentiation of mesenchymal stem cells via targeting HDAC4. Eur Rev Med Pharmacol Sci. 2018;22(11):3318–3326.
- Wu S, Yu Q, Lai A, Tian J. Pulsed electromagnetic field induces Ca(2+)-dependent osteoblastogenesis in C3H10T1/2 mesenchymal cells through the Wnt-Ca(2+)/Wnt-beta-catenin signaling pathway. Biochem Biophys Res Commun. 2018;503(2):715–721.
- Mansouri R, Jouan Y, Hay E, et al. Osteoblastic heparan sulfate glycosaminoglycans control bone remodeling by regulating Wnt signaling and the crosstalk between bone surface and marrow cells. Cell Death Dis. 2017;8(6):e2902.
- Blitzer JT. Nusse R. A critical role for endocytosis in Wnt signaling. BMC Cell Biol. 2006;7:28.
- Delgado Cuenca P, Almaiman L, Schenk S, Kern M, Hooshmand S. Dried plum ingestion increases the osteoblastogenic capacity of human serum. J Med Food. 2017;20(7):653–658.
- John AA, Prakash R, Kureel J, Singh D. Identification of novel microRNA inhibiting actin cytoskeletal rearrangement thereby suppressing osteoblast differentiation. J Mol Med (Berl). 2018;96(5):427–444.
- Tan J, Tong BD, Wu YJ, Xiong W. MicroRNA-29 mediates TGFβ1-induced extracellular matrix synthesis by targeting wnt/β-catenin pathway in human orbital fibroblasts. Int J Clin Exp Pathol. 2014;7(11):7571–7577.
- Luyten FP, Tylzanowski P, Lories RJ. Wnt signaling and osteoarthritis. Bone. 2009;44(4):522–527.
- Clevers H, Nusse R. Wnt/beta-catenin signaling and disease. Cell. 2012;149(6):1192–1205.
- Weng LH, Wang CJ, Ko JY, Sun YC, Wang FS. Control of Dkk‐1 ameliorates chondrocyte apoptosis, cartilage destruction and subchondral bone deterioration in osteoarthritic knees. Arthritis Rheum. 2010;62(5):1393–1402.
- Kapinas K, Kessler CB, Delany AM. MiR-29 suppression of osteonectin in osteoblasts: Regulation during differentiation and by canonical Wnt signaling. J Cell Biochem. 2009;108(1):216–224.
- Laine CM, Joeng KS, Campeau PM, et al. WNT1 mutations in early-onset osteoporosis and osteogenesis imperfecta. N Engl J Med. 2013;368(19):1809–1816.
- Moorer MC, Riddle RC. Regulation of osteoblast metabolism by Wnt signaling. Endocrinol Metab (Seoul). 2018;33(3):318–330.
- Hill TP, Spater D, Taketo MM, Birchmeier W, Hartmann C. Canonical Wnt/beta-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell. 2005;8(5):727–738.