miR-129-1-3p通过BMP2/SMAD1信号通路抑制人骨髓间充质干细胞成骨分化的研究

doi:10.13591/j.cnki.kqyx.2025.06.004

Abstract

Abstract:

Objective To investigate the effect of miR-129-1-3p on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) and its potential mechanism. Methods Negative control group, the miR-129-1-3p mimic group, the miR-129-1-3p inhibitor group and the corresponding negative control were constructed and transfected into hBMSCs. The formation of calcium-mineralized nodules was observed by alkaline phosphatase staining and alizarin red S staining. The expression levels of miR-129-1-3p and osteogenic differentiation markers were detected by qRT-PCR. Western blot detected the protein expressions of bone morphogenetic protein 2 (BMP2), SMAD1 and p-SMAD1. Results After transfection, the expression level of miR-129-1-3p in mimic group was significantly increased (P<0.05), the number of mineralized nodules was significantly decreased, and the expression levels of BMP2, Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) mRNA were significantly down-regulated (P<0.05). BMP2 and p-SMAD1 protein were also significantly down-regulated (P<0.05) compared with Mimic-NC group. The expression levels of BMP2, RUNX2, OCN mRNA were significantly up-regulated in inhibitor group (P<0.05) compared with Inhibitor-NC group. BMP2 and p-SMAD1 protein were significantly up-regulated in inhibitor group (P<0.05). Conclusion miR-129-1-3p can inhibit the osteogenic differentiation of human bone marrow mesenchymal stem cells by suppressing BMP2/SMAD1 signaling pathway.

Key words: miR-129-1-3p, BMP2/SMAD1 signaling pathway, human bone marrow mesenchymal stem cells, osteogenic differentiation, osteoporosis

CLC Number:

R780.2

GENG Mingzhu, MU Wenqing, QIU Lin, ZHANG Wei. miR-129-1-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells via BMP2/SMAD1 signaling pathway[J]. Stomatology, 2025, 45(6): 418-423.

Figures/Tables 7

Tab.1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

References 37

[1]	Ensrud KE, Crandall CJ. Osteoporosis[J]. Ann Intern Med, 2024, 177(1): ITC1-ITC16.
[2]	Lane NE. Epidemiology, etiology, and diagnosis of osteoporosis[J]. Am J Obstet Gynecol, 2006, 194(2 Suppl): S3-11. doi: 10.1016/j.ajog.2005.08.047 pmid: 16448873
[3]	Ramchand SK, Leder BZ. Sequential therapy for the long-term treatment of postmenopausal osteoporosis[J]. J Clin Endocrinol Metab, 2024, 109(2): 303-311.
[4]	Pittenger MF, MacKay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells[J]. Science, 1999, 284(5411): 143-147. doi: 10.1126/science.284.5411.143 pmid: 10102814
[5]	Gao Y, Chen N, Fu Z, et al. Progress of Wnt signaling pathway in osteoporosis[J]. Biomolecules, 2023, 13(3): 483.
[6]	Bonor J, Adams EL, Bragdon B, et al. Initiation of BMP2 signaling in domains on the plasma membrane[J]. J Cell Physiol, 2012, 227(7): 2880-2888. doi: 10.1002/jcp.23032 pmid: 21938723
[7]	Wang Z, Bao HW, Xu YJ. Cnidium lactone prevents bone loss in an ovariectomized rat model through the estrogen-α/BMP-2/Smad signaling pathway[J]. J Gene Med, 2020, 22(8): e3198.
[8]	Lu TX, Rothenberg ME. microRNA[J]. J Allergy Clin Immunol, 2018, 141(4): 1202-1207. doi: S0091-6749(17)31593-2 pmid: 29074454
[9]	Fesler A, Zhai H, Ju J. miR-129 as a novel therapeutic target and biomarker in gastrointestinal cancer[J]. Onco Targets Ther, 2014, 7: 1481-1485.
[10]	Fu R, Yang P, Sajid A, et al. Avenanthramide A induces cellular senescence miR-129-3p/Pirh2/p53 signaling pathway to suppress colon cancer growth[J]. J Agric Food Chem, 2019, 67(17): 4808-4816.
[11]	Ma L, Chen X, Li C, et al. miR-129-5p and-3p co-target WWP1 to suppress gastric cancer proliferation and migration[J]. J Cell Biochem, 2019, 120(5): 7527-7538.
[12]	Wu YF, Ou CC, Chien PJ, et al. Chidamide-induced ROS accumulation and miR-129-3p-dependent cell cycle arrest in non-small lung cancer cells[J]. Phytomedicine, 2019, 56: 94-102.
[13]	Li Q, Qin M, Tan Q, et al. MicroRNA-129-1-3p protects cardiomyocytes from pirarubicin-induced apoptosis by down-regulating the GRIN2D-mediated Ca²⁺ signalling pathway[J]. J Cell Mol Med, 2020, 24(3): 2260-2271. doi: 10.1111/jcmm.14908 pmid: 31957170
[14]	Yin C, Tian Y, Yu Y, et al. miR-129-5p inhibits bone formation through TCF4[J]. Front Cell Dev Biol, 2020, 8: 600641.
[15]	Zhao C, Gu Y, Wang Y, et al. miR-129-5p promotes osteogenic differentiation of BMSCs and bone regeneration repressing Dkk3[J]. Stem Cells Int, 2021, 2021(1): 7435605.
[16]	Chen R, Ye B, Xie H, et al. miR-129-3p alleviates chondrocyte apoptosis in knee joint fracture-induced osteoarthritis through CPEB1[J]. J Orthop Surg Res, 2020, 15(1): 552. doi: 10.1186/s13018-020-02070-1 pmid: 33228708
[17]	Reid IR. A broader strategy for osteoporosis interventions[J]. Nat Rev Endocrinol, 2020, 16(6): 333-339. doi: 10.1038/s41574-020-0339-7 pmid: 32203407
[18]	Ayers C, Kansagara D, Lazur B, et al. Effectiveness and safety of treatments to prevent fractures in people with low bone mass or primary osteoporosis: A living systematic review and network meta-analysis for the American college of physicians[J]. Ann Intern Med, 2023, 176(2): 182-195.
[19]	Zhao Y, He JW, Qiu T, et al. Epigenetic therapy targeting bone marrow mesenchymal stem cells for age-related bone diseases[J]. Stem Cell Res Ther, 2022, 13(1): 201. doi: 10.1186/s13287-022-02852-w pmid: 35578312
[20]	Pérez-Campo FM, Santurtún A, García-Ibarbia C, et al. Osterix and RUNX2 are transcriptional regulators of sclerostin in human bone[J]. Calcif Tissue Int, 2016, 99(3): 302-309.
[21]	Gori F, Thomas T, Hicok KC, et al. Differentiation of human marrow stromal precursor cells:Bone morphogenetic protein-2 increases OSF2/CBFA1, enhances osteoblast commitment, and inhibits late adipocyte maturation[J]. J Bone Miner Res, 1999, 14(9): 1522-1535. pmid: 10469280
[22]	Bialek P, Kern B, Yang X, et al. A twist code determines the onset of osteoblast differentiation[J]. Dev Cell, 2004, 6(3): 423-435. doi: 10.1016/s1534-5807(04)00058-9 pmid: 15030764
[23]	Yi X, Wright LE, Pagnotti GM, et al. Mechanical suppression of breast cancer cell invasion and paracrine signaling to osteoclasts requires nucleo-cytoskeletal connectivity[J]. Bone Res, 2020, 8(1): 40. doi: 10.1038/s41413-020-00111-3 pmid: 33298883
[24]	Haffner-Luntzer M, Kovtun A, Lackner I, et al. Estrogen receptor α-(ERα), but not ERβ-signaling, is crucially involved in mechanostimulation of bone fracture healing by whole-body vibration[J]. Bone, 2018, 110: 11-20. doi: S8756-3282(18)30017-6 pmid: 29367057
[25]	Fang S, He T, You M, et al. Glucocorticoids promote steroid-induced osteonecrosis of the femoral head by down-regulating serum alpha-2-macroglobulin to induce oxidative stress and facilitate SIRT2-mediated BMP2 deacetylation[J]. Free Radic Biol Med, 2024, 213: 208-221.
[26]	Luu YK, Capilla E, Rosen CJ, et al. Mechanical stimulation of mesenchymal stem cell proliferation and differentiation promotes osteogenesis while preventing dietary-induced obesity[J]. J Bone Miner Res, 2009, 24(1): 50-61.
[27]	Diener C, Keller A, Meese E. Emerging concepts of miRNA therapeutics: From cells to clinic[J]. Trends Genet, 2022, 38(6): 613-626. doi: 10.1016/j.tig.2022.02.006 pmid: 35303998
[28]	Rupaimoole R, Slack FJ. MicroRNA therapeutics:Towards a new era for the management of cancer and other diseases[J]. Nat Rev Drug Discov, 2017, 16(3): 203-222. doi: 10.1038/nrd.2016.246 pmid: 28209991
[29]	Zhang Z, Wang M, Zheng Y, et al. MicroRNA-223 negatively regulates the osteogenic differentiation of periodontal ligament derived cells by directly targeting growth factor receptors[J]. J Transl Med, 2022, 20(1): 465. doi: 10.1186/s12967-022-03676-1 pmid: 36221121
[30]	Cao Y, Lv Q, Lv C. MicroRNA-153 suppresses the osteogenic differentiation of human mesenchymal stem cells by targeting bone morphogenetic protein receptor type Ⅱ[J]. Int J Mol Med, 2015, 36(3): 760-766.
[31]	Marupanthorn K, Tantrawatpan C, Kheolamai P, et al. MicroRNA treatment modulates osteogenic differentiation potential of mesenchymal stem cells derived from human chorion and placenta[J]. Sci Rep, 2021, 11(1): 7670. doi: 10.1038/s41598-021-87298-5 pmid: 33828198
[32]	Maeda K, Kobayashi Y, Koide M, et al. The regulation of bone metabolism and disorders by Wnt signaling[J]. Int J Mol Sci, 2019, 20(22): 5525.
[33]	Zieba JT, Chen YT, Lee BH, et al. Notch Signaling in Skeletal Development, Homeostasis and Pathogenesis[J]. Biomolecules, 2020, 10(2): 332.
[34]	Chen G, Deng C, Li YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation[J]. Int J Biol Sci, 2012, 8(2): 272-288. doi: 10.7150/ijbs.2929 pmid: 22298955
[35]	Salazar VS, Gamer LW, Rosen V. BMP signalling in skeletal development, disease and repair[J]. Nat Rev Endocrinol, 2016, 12(4): 203-221. doi: 10.1038/nrendo.2016.12 pmid: 26893264
[36]	Li Y, Chen G, He Y, et al. Selenomethionine-modified polyethylenimine-based nanoparticles loaded with miR-132-3p inhibitor-biofunctionalized titanium implants for improved osteointegration[J]. ACS Biomater Sci Eng, 2021, 7(10): 4933-4945.
[37]	Rojo Arias JE, Busskamp V. Challenges in microRNAs’ targetome prediction and validation[J]. Neural Regen Res, 2019, 14(10): 1672-1677.

基因名称	引物序列(5'→3')
RUNX2-F	GACGAGGCAAGAGTTTCACC
RUNX2-R	GGTTCCCGAGGTCCATCTAC
BMP2-F	TTCGGCCTGAAACAGAGACC
BMP2-R	CCTGAGTGCCTGCGATACAG
OCN-F	CTACCTGTATCAA
OCN-R	GGATTGAGCTCACACACCT
miR-129-1-3p RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCA-CTGGATACGACATACTT
miR-129-1-3p-F	ACCATTCAAAGCCCTTACCCCAA
miR-129-1-3p-R	ATCCAGTGCAGGGTCCGAGG
GAPDH-F	CAATGACCCCTTCATTGACC
GAPDH-R	TTGATTTTGGAGGGATCTCG

miR-129-1-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells via BMP2/SMAD1 signaling pathway

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 37

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	DONG Weijie, SU Tingshu, XIN Xianzhen. Effects of quercetin loaded gelatin microspheres on proliferation and differentiation of MC3T3-E1 [J]. Stomatology, 2024, 44(7): 494-499.
[2]	PAN Le, DUAN Qinyan, CHENG Junxiang, HONG Feng, HU Yajun. Research progress of LncRNA-mediated miRNA regulating osteogenic differentiation of PDLSCs in microenvironment [J]. Stomatology, 2024, 44(3): 232-236.
[3]	DING Kexin, YANG Jinxin, MOU Jie, SUN Zhe, CUI Yawen, LIU Zongxiang. In vitro study on flavonoid NO donor nanoparticles promoting osteogenic differentiation of PDLSCs by regulating macrophage polarization [J]. Stomatology, 2024, 44(11): 806-814.
[4]	LUO Guisheng, GU Teng, LI Junjun, WANG Penglai, ZHANG Cheng, YUAN Changyong. Effect of melatonin-modified PEEK implant on osteointegration in osteoporotic rats [J]. Stomatology, 2024, 44(10): 734-741.
[5]	JIANG Zhen, HAN Xue, JIANG Beizhan. Research progress of the regulatory mechanisms of dental follicle in bone remodeling during tooth eruption [J]. Stomatology, 2023, 43(8): 752-756.
[6]	GE Xiao,YU Miao,WU Wei,BI Xiuting,WU Xiaoyan,YU Chen,LI Ti. The effect of tantalum coating on the proliferation and osteogenic differentiation of human periodontal ligament stem cells [J]. Stomatology, 2023, 43(5): 415-420.
[7]	LIU Yuqing, GONG Yao, ZHAO Sufeng, DAI Li. Regulation effect of miR-155 on osteogenic differentiation of periodontal ligament stem cells in inflammatory microenvironment [J]. Stomatology, 2022, 42(9): 790-795.
[8]	ZHANG Shuting, WU Yaxing, LIU Cuicui, LI Chenchen, ZHANG Jing. Research progress of exosomes in osteogenic differentiation of bone marrow mesenchymal stem cells and in periodontal regeneration [J]. Stomatology, 2022, 42(8): 741-744.
[9]	ZHANG Xiaochen, SUN Weifu, FANG Shishu, QIN Wen, JIN Zuolin. Research progress of N⁶-methyladenosine in regulating osteogenic differentiation [J]. Stomatology, 2022, 42(7): 655-658.
[10]	SHA Sha, LU Lingbo, GUO Peipei, CHEN Shu, JIN Muhan, XU Rongyao. Clinical study on correlation between miRNA-143/145 in saliva and estrogen-deficient osteoporosis [J]. Stomatology, 2022, 42(6): 515-520.
[11]	LI Yanyan, ZHU Zhu, XIE Wenjing, XU Zi′ang, ZHANG Ziwei, ZHANG Wei. The effect of asperosaponinⅥ on human jaw bone marrow mesenchymal stem cells' osteogenic differentiation [J]. Stomatology, 2022, 42(3): 204-209.
[12]	DONG Shuo, TANG Chunbo. Progress of research on the mechanism of PI3K/AKT signaling pathway in implant osseointegration in patients with type 2 diabetes mellitus [J]. Stomatology, 2022, 42(11): 1026-1030.
[13]	ZHANG Binjing, ZHANG Jinglan, ZHANG Chenyue, CHEN Yifei, HU Zhi′ai. Progress of research on the role of B lymphocytes in bone destructive diseases [J]. Stomatology, 2022, 42(10): 922-927.
[14]	Zhu Zhu. Effects of Asperosaponin Ⅵ on osteogenic differentiation of mouse myoblasts [J]. , 2021, 41(5): 398-402.
[15]	. Canonical Wnt signaling pathway and osteogenic differentiation of periodontal ligament stem cells [J]. , 2021, 41(10): 936-941.