|
|
|
| BMP9-induced Osteogenic Differentiation is Partially Inhibited by miR-30a in Mesenchymal Stem Cell Line C3H10 T1/2 |
| LI Bao-lin, BAI Hui-Li, ZHANG Ru-Yi, YAN Shu-Juan, HE-Fang, YANG Dang-Dang, LIU-Chen, SHI Qiong |
| Key Laboratory of Clinical Laboratory Diagnostics of Ministry Education, Faculty of Laboratory Medicine, Chongqing Medicine University, Chongqing 400016, China |
|
|
|
Abstract Objective:To study the role of miR-30a in bone morphogenetic protein 9 (BMP9) induced osteogenic differentiation of mesenchymal stem cell line C3H10T1/2.Method:C3H10T1/2 cells were treated with Adenovirus GFP and BMP9,the early osteogenic marker Alkaline phosphatase(ALP) activity was detected by ALP quantitative assay and staining assay,the later osteogenic marker calcium precipitation was detected by Alizarin Red S staining,OPN was another later marker,which was detected by PCR and Western blot.The expression of miR-30a in C3H10T1/2 cells quantified by Real-time PCR.Effects of miR-30a on BMP9-induced osteogenic differentiation of C3H10T1/2 cells was detected by ALP,OPN and calcium precipitation.Runx2 was detected in order to study its mechanism. Results: BMP9 was not only increased the activity of ALP on days 5,7,the calcium precipitation on 14 day was also increased compared with Ad-GFP group.miR-30a reduced the expression of ALP on BMP9-induced osteogenic differentiation.The expression of OPN and calcium precipitation were also inhibited by miR-30a.Conclusion:BMP9-induced osteogenic differentiation is partially inhibited by miR-30a in mesenchymal stem cell line C3H10 T1/2.
|
|
Received: 16 July 2013
Published: 25 November 2013
|
|
|
|
|
[1] Luu H H, Song W X, Luo X, et al. Distinct roles of bone morphogenetic proteins in osteogenic differentiation of mesenchymal stem cells.J Orthop Res, 2007, 25(5): 665-677.
[2] Kang Q, Sun M H, Cheng H, et al. Characterization of the distinct orthotopic bone forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Ther, 2004, 11 (17):1312-1320.
[3] O'Donnell K A, Wentzel E A, Zeller K I, et al.c-Myc-regulated microRNAs modulate E2F1 expression.Nature, 2005, 435(7043):839-843.
[4] Li Z, Hassan M Q, Volinia S, et al.A microRNA signature for a BMP2-induced osteoblast lineage commitment program.Proc Natl Acad Sci, 2008, 105(37)13906–13911.
[5] Bae Y, Yang T, Zeng H C, et al.miRNA-34c regulates Notch signaling during bone development.Hum Mol Genet, 2012, 21(13):2991-3000.
[6] Itoh T, Nozawa Y, Akao Y, et al.MicroRNA-141 and -200a are involved in bone morphogenetic protein-2-induced mouse pre-osteoblast differentiation by targeting distal-less homeobox 5.J Biol Chem, 2009, 284(29):19272-19279.
[7] Luo M, Weng Y, Tang J, et al.MicroRNA-450a-3p represses cell proliferation and regulates embryo development by regulating Bub1 expression in mouse.PLos One, 2012, 7(10):e47194
[8] Wu T, Zhou H, Hong Y, et al.miR-30 family members negatively regulate osteoblast differentiation.J Biol Chem, 2012, 287(10):7503-7511.
[9] Zaragosi L E, Wdziekonski B, Brigand K L, et al.Small RNA sequencing reveals miR-642a-3p as a novel adipocyte-specific microRNA and miR-30 as a key regulator of human adipogenesis.Genome Biol, 2011, 12(7):R64.
[10] 张燕, 文巍, 罗进勇. 骨形态发生蛋白9定向诱导多潜能干细胞成骨分化. 生物化学与生物物理进展, 2009, 36(10):12911-12298. Zhang Y, Wen W, Luo J Y, et al.BMP9 induces osteogenic differentiation of pluripotent stem cell. Prog Biochem Biophys, 2009, 36(10):12911-12298.
[11] Xu D J, Zhao Y Z, Wang J, et al.Smads, p38 and ERK1, 2 are involved in BMP9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.BMB Rep, 2012, 45(4):247-252.
[12] Kim Y J, Bae S W, Yu S S, et al.miR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue.J Bone Miner Res, 2009, 24(5):816-825.
[13] Eskildsen T, Yaipaleenmaki H, Stenvang J, et al.MicroRNA-138 regulates osteogenic differentiation of human stromal (mesenchymal) stem cells in vivo.Proc Natl Acad Sci, 2011, 108(15):6139-6144.
[14] Bhushan R, Grunhagen J, Becker J, et al.miR-181a promotes osteoblastic differentiation through repression of TGF-β signaling molecules.Int J Biochem Cell Biol, 2013, 45(3):696-705.
[15] 赵丹, 罗进勇. BMP9促进间充质干细胞C3H10T1/2成骨分化的研究. 临床和实验医学杂志, 2011, 10(16): 1225-1230. Zhao D, Luo J Y.The promotive effect of BMP9 in osteogenic differentiation of C3H10T1/2 mesenchymal stem cells. Journal of Clinical and Experimental Medicine, 2011, 10(16): 1225-1230.
[16] Zaragosi L E, Wdziekonski B, Brigand K L, et al.Small RNA sequencing reveals miR-642a-3p as a novel adipocyte-specific microRNA and miR-30 as a key regulator of human adipogenesis.Genome Bio, 2011, 12(7):R64.
[17] Komori T. Mechanism of transcriptional regulation by Runx2 in osteoblasts. Clin Calcium, 2006, 16(5):801-807.
[18] Phimphilai M, Zhao Z, Boules H, et al. BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res, 2006, 21(4):637-646.
[19] Enomoto H, Furuichi T, Zanmana A, et al.Runx2 deficiency in chondrocytes causes adipogenic changes in vitro.J Cell Sci, 2004, 117(Pt3):417-425. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
