Notch‑1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells

He,Ying; Zou,Lijin

doi:10.3892/etm.2019.7765

Notch‑1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells

Authors:
- Ying He
- Lijin Zou
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Affiliations: Department of Infectious Diseases, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Burn Center, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Published online on: July 10, 2019 https://doi.org/10.3892/etm.2019.7765
Pages: 1884-1890

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Abstract

Low differentiation and high proliferation rates are critical factors affecting bone marrow mesenchymal stem cell (BMSC) tumorigenesis. The present study aimed to investigate the role of the Notch signaling pathway in BMSC proliferation and osteogenic differentiation. Mouse BMSCs were divided into control, vector, Notch1‑small interfering (si)RNA, γ‑secretase inhibitor, and Notch1‑siRNA + γ‑secretase inhibitor groups. The siRNA‑Notch1, γ‑secretase inhibitor, and Notch1‑siRNA + γ‑secretase inhibitor groups were treated with Notch1 siRNA and/or γ‑secretase inhibitor. Following treatment, cell proliferation was evaluated using a Cell Counting Kit‑8. Tumor‑related factors, including transforming growth factor (TGF)‑β1, c‑Myc and p53, were detected by reverse transcription‑quantitative polymerase chain reaction and western blot analyses. BMSC osteogenic differentiation was induced and the cells were stained with alizarin red at 14 and 21 days. Alkaline phosphatase (AKP) activity was also evaluated. The siRNA‑Notch1 and γ‑secretase inhibitor both reduced BMSC proliferation and the expression of TGF‑β1 and c‑Myc and increased the expression of p53. Following the induction of osteogenesis and staining with alizarin red, the level of AKP was significantly higher in cells in the siRNA‑Notch1 and γ‑secretase inhibitor groups compared with that in the control group. It was found that Notch1 inhibition reduced proliferation and promoted the osteogenic differentiation of BMSCs.

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1	Ren T, Piperdi S, Koirala P, Park A, Zhang W, Ivenitsky D, Zhang Y, Villanueva-Siles E, Hawkins DS, Roth M and Gorlick R: CD49b inhibits osteogenic differentiation and plays an important role in osteosarcoma progression. Oncotarget. 8:87848–87859. 2017. View Article : Google Scholar : PubMed/NCBI
2	Nombela-Arrieta C, Ritz J and Silberstein LE: The elusive nature and function of mesenchymal stem cells. Nat Rev Mol Cell Biol. 12:126–131. 2011. View Article : Google Scholar : PubMed/NCBI
3	Williams AR and Hare JM: Mesenchymal stem cells: Biology, pathophysiology, translational findings, and therapeutic implications for cardiac disease. Circ Res. 109:923–940. 2011. View Article : Google Scholar : PubMed/NCBI
4	Morancho A, Ma F, Barcelo V, Giralt D, Montaner J and Rosell A: Impaired vascular remodeling after endothelial progenitor cell transplantation in MMP9-deficient mice suffering cortical cerebral ischemia. J Cereb Blood Flow Metab. 35:1547–1551. 2015. View Article : Google Scholar : PubMed/NCBI
5	Wallner C, Abraham S, Wagner JM, Harati K, Ismer B, Kessler L, Zöllner H, Lehnhardt M and Behr B: Local application of isogenic adipose-derived stem cells restores bone healing capacity in a type 2 diabetes model. Stem Cell Transl Med. 5:836–844. 2016. View Article : Google Scholar
6	Gupta PK, Chullikana A, Parakh R, Desai S, Das A, Gottipamula S, Krishnamurthy S, Anthony N, Pherwani A and Majumdar AS: A double blind randomized placebo controlled phase I/II study assessing the safety and efficacy of allogeneic bone marrow derived mesenchymal stem cell in critical limb ischemia. J Transl Med. 11:1432013. View Article : Google Scholar : PubMed/NCBI
7	Lara-Hernandez R, Lozano-Vilardell P, Blanes P, Torreguitart-Mirada N, Galmes A and Besalduch J: Safety and efficacy of therapeutic angiogenesis as a novel treatment in patients with critical limb ischemia. Ann Vasc Surg. 24:287–294. 2010. View Article : Google Scholar : PubMed/NCBI
8	He Y, Zhu W, Shin MH, Gary J, Liu C, Dubois W, Hoover SB, Jiang S, Marrogi E, Mock B, et al: cFOS-SOX9 axis reprograms bone marrow-derived mesenchymal stem cells into chondroblastic osteosarcoma. Stem Cell Rep. 8:1630–1644. 2017. View Article : Google Scholar
9	Yuan TM and Yu HM: Notch signaling: Key role in intrauterine infection/inflammation, embryonic development, and white matter damage? J Neurosci Res. 88:461–468. 2010.PubMed/NCBI
10	Reddy BV, Rauskolb C and Irvine KD: Influence of fat-hippo and notch signaling on the proliferation and differentiation of Drosophila optic neuroepithelia. Development. 137:2397–2408. 2010. View Article : Google Scholar : PubMed/NCBI
11	Bray SJ: Notch signalling in context. Nat Rev Mol Cell Biol. 17:722–735. 2016. View Article : Google Scholar : PubMed/NCBI
12	Colombo M, Galletti S, Garavelli S, Platonova N, Paoli A, Basile A, Taiana E, Neri A and Chiaramonte R: Notch signaling deregulation in multiple myeloma: A rational molecular target. Oncotarget. 6:26826–26840. 2015. View Article : Google Scholar : PubMed/NCBI
13	Cristofaro B and Emanueli C: Possible novel targets for therapeutic angiogenesis. Curr Opin Pharmacol. 9:102–108. 2009. View Article : Google Scholar : PubMed/NCBI
14	Liang T, Zhu L, Gao W, Gong M, Ren J, Yao H, Wang K and Shi D: Coculture of endothelial progenitor cells and mesenchymal stem cells enhanced their proliferation and angiogenesis through PDGF and Notch signaling. FEBS Open Bio. 7:1722–1736. 2017. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Gibbs C, Kukekov VG, Reith JD, Tchigrinova O, Suslov ON, Scott EW, Ghivizzani SC, Ignatova TN and Steindler DA: Stem-like cells in bone sarcomas: Implications for tumorigenesis. Neoplasia. 7:967–976. 2005. View Article : Google Scholar : PubMed/NCBI
17	Berman SD, Calo E, Landman AS, Danielian PS, Miller ES, West JC, Fonhoue BD, Caron A, Bronson R, Bouxsein ML, et al: Metastatic osteosarcoma induced by inactivation of Rb and p53 in the osteoblast lineage. Proc Natl Acad Sci USA. 105:11851–11856. 2008. View Article : Google Scholar : PubMed/NCBI
18	Tataria M, Quarto N, Longaker MT and Sylvester KG: Absence of the p53 tumor suppressor gene promotes osteogenesis in mesenchymal stem cells. J Pediatr Surg. 41:624–632. 2006. View Article : Google Scholar : PubMed/NCBI
19	Sun Z, Wang S and Zhao RC: The roles of mesenchymal stem cells in tumor inflammatory microenvironment. J Hematol Oncol. 7:142014. View Article : Google Scholar : PubMed/NCBI
20	Hill BS, Pelagalli A, Passaro N and Zannetti A: Tumor-educated mesenchymal stem cells promote pro-metastatic phenotype. Oncotarget. 8:73296–73311. 2017. View Article : Google Scholar : PubMed/NCBI
21	Cao J, Wei Y, Lian J, Yang L, Zhang X, Xie J, Liu Q, Luo J, He B and Tang M: Notch signaling pathway promotes osteogenic differentiation of mesenchymal stem cells by enhancing BMP9/Smad signaling. Int J Mol Med. 40:378–388. 2017. View Article : Google Scholar : PubMed/NCBI
22	Cheng Y, Gu W, Zhang G, Li X and Guo X: Activation of Notch1 signaling alleviates dysfunction of bone marrow-derived mesenchymal stem cells induced by cigarette smoke extract. Int J Chron Obstruct Pulmon Dis. 12:3133–3147. 2017. View Article : Google Scholar : PubMed/NCBI
23	Li TS, Hayashi M, Ito H, Furutani A, Murata T, Matsuzaki M and Hamano K: Regeneration of infarcted myocardium by intramyocardial implantation of ex vivo transforming growth factor-beta-preprogrammed bone marrow stem cells. Circulation. 111:2438–2445. 2005. View Article : Google Scholar : PubMed/NCBI
24	Li Y, Powell S, Brunette E, Lebkowski J and Mandalam R: Expansion of human embryonic stem cells in defined serum-free medium devoid of animal-derived products. Biotechnol Bioeng. 91:688–698. 2005. View Article : Google Scholar : PubMed/NCBI
25	Gwak SJ, Bhang SH, Yang HS, Kim SS, Lee DH, Lee SH and Kim BS: In vitro cardiomyogenic differentiation of adipose-derived stromal cells using transforming growth factor-beta1. Cell Biochem Funct. 27:148–154. 2009. View Article : Google Scholar : PubMed/NCBI
26	Akhurst RJ, Lehnert SA, Faissner A and Duffie E: TGF beta in murine morphogenetic processes: The early embryo and cardiogenesis. Development. 108:645–656. 1990.PubMed/NCBI
27	Chen S and Lechleider RJ: Transforming growth factor-beta-induced differentiation of smooth muscle from a neural crest stem cell line. Circ Res. 94:1195–1202. 2004. View Article : Google Scholar : PubMed/NCBI
28	Mueller MB, Fischer M, Zellner J, Berner A, Dienstknecht T, Prantl L, Kujat R, Nerlich M, Tuan RS and Angele P: Hypertrophy in mesenchymal stem cell chondrogenesis: Effect of TGF-beta isoforms and chondrogenic conditioning. Cells Tissues Organs. 192:158–166. 2010. View Article : Google Scholar : PubMed/NCBI
29	Hu ML, Wang XY and Chen WM: TGF-β1 upregulates the expression of lncRNA UCA1 and its downstream HXK2 to promote the growth of hepatocellular carcinoma. Eur Rev Med Pharmacol Sci. 22:4846–4854. 2018.PubMed/NCBI
30	Viloria ME, Bravo J, Carrero Y and Mosquera JA: In situ expressions of protein 16 (p16^CDKN2A)) and transforming growth factor beta-1 in patients with cervical intraepithelial neoplasia and cervical cancer. Eur J Obstet Gynecol Reprod Biol. 228:303–307. 2018. View Article : Google Scholar : PubMed/NCBI
31	Cui M, Chang Y, Du W, Liu S, Qi J, Luo R and Luo S: Upregulation of lncRNA-ATB by transforming growth factor beta1 (TGF-β1) promotes migration and invasion of papillary thyroid carcinoma cells. Med Sci Monit. 24:5152–5158. 2018. View Article : Google Scholar : PubMed/NCBI
32	Gallant P: Myc/Max/Mad in invertebrates: The evolution of the Max network. Curr Top Microbiol Immunol. 302:235–253. 2006.PubMed/NCBI
33	Lin TP, Li J, Li Q, Li X, Liu C, Zeng N, Huang JM, Chu GC, Lin CH, Zhau HE, et al: R1 Regulates prostate tumor growth and progression by transcriptional suppression of the E3 Ligase HUWE1 to Stabilize c-Myc. Mol Cancer Res. 16:1940–1951. 2018. View Article : Google Scholar : PubMed/NCBI
34	Zhou X, Wen Y, Tian Y, He M, Ke X, Huang Z, He Y, Liu L, Scharf A, Lu M, et al: Hsp90alpha-dependent B-Claf-1 promotes hepatocellular carcinoma proliferation by regulating c-MYC mRNA stability. Hepatology. 2018.
35	Zanotti S, Smerdel-Ramoya A, Stadmeyer L, Durant D, Radtke F and Canalis E: Notch inhibits osteoblast differentiation and causes osteopenia. Endocrinology. 149:3890–3899. 2008. View Article : Google Scholar : PubMed/NCBI