Role of p53 in promoting BMP9‑induced osteogenic differentiation of mesenchymal stem cells through TGF‑β1

Yao,Xintong; Li,Peipei; Deng,Yixuan; Yang,Yuanyuan; Luo,Honghong; He,Baicheng

doi:10.3892/etm.2023.11947

Role of p53 in promoting BMP9‑induced osteogenic differentiation of mesenchymal stem cells through TGF‑β1

Authors:
- Xintong Yao
- Peipei Li
- Yixuan Deng
- Yuanyuan Yang
- Honghong Luo
- Baicheng He
View Affiliations

Affiliations: College of Pharmacy, Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: April 13, 2023 https://doi.org/10.3892/etm.2023.11947
Article Number: 248
Copyright: © Yao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Known as a tumour suppressor gene, p53 also plays a key role in controlling the differentiation of mesenchymal stem cells (MSCs). Bone morphogenetic protein 9 (BMP9) has been identified as a potent factor in inducing osteogenic differentiation of MSCs, but its relationship with p53 remains unclear. The present study revealed that TP53 was expressed at higher levels in MSCs from patients with osteoporosis and was associated with the top 10 core central genes found in the current osteoporosis genetic screen. p53 was expressed in C2C12, C3H10T1/2, 3T3‑L1, MEFs, and MG‑63 cell lines, and could be upregulated by BMP9, as measured by western blotting and reverse‑transcription quantitative PCR (RT‑qPCR). Furthermore, overexpression of p53 increased the mRNA and protein levels of osteogenic marker Runx2 and osteopontin, as evaluated by western blotting and RT‑qPCR in BMP9‑induced MSCs, whereas the p53 inhibitor pifithrin (PFT)‑α attenuated these effects. The same trend was found in alkaline phosphatase activities and matrix mineralization, as measured by alkaline phosphatase staining and alizarin red S staining. Moreover, p53 overexpression reduced adipo‑differentiation markers of PPARγ and lipid droplet formation, as measured by western blotting, RT‑qPCR and oil red O staining, respectively, whereas PFT‑α facilitated adipo‑differentiation in MSCs. In addition, p53 promoted TGF‑β1 expression and inhibition of TGF‑β1 by LY364947 partially attenuated the effects of p53 on promoting BMP9‑induced MSC osteo‑differentiation and inhibiting adipo‑differentiation. The inhibitory effect of PFT‑α on osteogenic markers and the promoting effect on adipogenic markers can be reversed when combined with TGF‑β1. TGF‑β1 may enhance the promotion of osteo‑differentiation of MSCs by p53 through inhibition of adipo‑differentiation. Collectively, by promoting BMP9‑induced MSCs bone differentiation and inhibiting adipose differentiation, p53 may be a novel therapeutic target for bone‑related diseases.

View Figures

View References

1	Pinheiro MB, Oliveira J, Bauman A, Fairhall N, Kwok W and Sherrington C: Evidence on physical activity and osteoporosis prevention for people aged 65+ years: A systematic review to inform the WHO guidelines on physical activity and sedentary behavior. Int J Behav Nutr Phys Act. 17(150)2020.PubMed/NCBI View Article : Google Scholar
2	Yang TL, Shen H, Liu A, Dong SS, Zhang L, Deng FY, Zhao Q and Deng HW: A road map for understanding molecular and genetic determinants of osteoporosis. Nat Rev Endocrinol. 16:91–103. 2020.PubMed/NCBI View Article : Google Scholar
3	Sözen T, Özışık L and Başaran NÇ: An overview and management of osteoporosis. Eur J Rheumatol. 4:46–56. 2017.PubMed/NCBI View Article : Google Scholar
4	Cheng C, Wentworth K and Shoback DM: New frontiers in osteoporosis therapy. Ann Rev Med. 71:277–288. 2020.PubMed/NCBI View Article : Google Scholar
5	Shao J, Zhang W and Yang T: Using mesenchymal stem cells as a therapy for bone regeneration and repairing. Biol Res. 48:1–7. 2015.PubMed/NCBI View Article : Google Scholar
6	Lu LX, Zhang XF, Wang YY, Ortiz L, Mao X, Jiang ZL, Xiao ZD and Huang NP: Effects of hydroxyapatite-containing composite nanofibers on osteogenesis of mesenchymal stem cells in vitro and bone regeneration in vivo. ACS Appl Mater Interfaces. 5:319–330. 2013.PubMed/NCBI View Article : Google Scholar
7	Wu M, Chen G and Li YP: TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Res. 4:1–21. 2016.PubMed/NCBI View Article : Google Scholar
8	Mostafa S, Pakvasa M, Coalson E, Zhu A, Alverdy A, Castillo H, Fan J, Li A, Feng Y, Wu D, et al: The wonders of BMP9: From mesenchymal stem cell differentiation, angiogenesis, neurogenesis, tumorigenesis, and metabolism to regenerative medicine. Genes Dis. 6:201–223. 2019.PubMed/NCBI View Article : Google Scholar
9	Wang Y, Ma C, Sun T and Ren L: Potential roles of bone morphogenetic protein-9 in glucose and lipid homeostasis. J Physiol Biochem. 76:503–512. 2020.PubMed/NCBI View Article : Google Scholar
10	Liu Y and Gu W: The complexity of p53-mediated metabolic regulation in tumor suppression. Semin Cancer Biol. 85:4–32. 2021.PubMed/NCBI View Article : Google Scholar
11	Lavin MF and Gueven N: The complexity of p53 stabilization and activation. Cell Death Differ. 13:941–950. 2006.PubMed/NCBI View Article : Google Scholar
12	Liu Y, Tavana O and Gu W: p53 modifications: Exquisite decorations of the powerful guardian. J Mol Cell Biol. 11:564–577. 2019.PubMed/NCBI View Article : Google Scholar
13	Vousden KH: Outcomes of p53 activation-spoilt for choice. J Cell Sci. 119:5015–5020. 2006.PubMed/NCBI View Article : Google Scholar
14	Huang Q, Liu M, Du X, Zhang R, Xue Y, Zhang Y, Zhu W, Li D, Zhao A and Liu Y: Role of p53 in preadipocyte differentiation. Cell Biol Int. 38:1384–1393. 2014.PubMed/NCBI View Article : Google Scholar
15	Jain AK, Allton K, Iacovino M, Mahen E, Milczarek RJ, Zwaka TP, Kyba M and Barton MC: p53 regulates cell cycle and microRNAs to promote differentiation of human embryonic stem cells. PLoS Biol. 10(e1001268)2012.PubMed/NCBI View Article : Google Scholar
16	Molchadsky A, Shats I, Goldfinger N, Pevsner-Fischer M, Olson M, Rinon A, Tzahor E, Lozano G, Zipori D, Sarig R and Rotter V: p53 plays a role in mesenchymal differentiation programs, in a cell fate dependent manner. PLoS One. 3(e3707)2008.PubMed/NCBI View Article : Google Scholar
17	Mao X, Li X, Hu W, Hao S, Yuan Y, Guan L and Guo B: Downregulated brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein-1 inhibits osteogenesis of BMSCs through p53 in type 2 diabetes mellitus. Biol Open. 9(bio051482)2020.PubMed/NCBI View Article : Google Scholar
18	Hüttinger-Kirchhof N, Cam H, Griesmann H, Hofmann L, Beitzinger M and Stiewe T: The p53 family inhibitor ΔNp73 interferes with multiple developmental programs. Cell Death Differ. 13:174–177. 2006.PubMed/NCBI View Article : Google Scholar
19	Velletri T, Huang Y, Wang Y, Li Q, Hu M, Xie N, Yang Q, Chen X, Chen Q, Shou P, et al: Loss of p53 in mesenchymal stem cells promotes alteration of bone remodeling through negative regulation of osteoprotegerin. Cell Death Differ. 28:156–169. 2021.PubMed/NCBI View Article : Google Scholar
20	Wang X, Kua HY, Hu Y, Guo K, Zeng Q, Wu Q, Ng HH, Karsenty G, de Crombrugghe B, Yeh J and Li B: p53 functions as a negative regulator of osteoblastogenesis, osteoblast-dependent osteoclastogenesis, and bone remodeling. J Cell Biol. 172:115–125. 2006.PubMed/NCBI View Article : Google Scholar
21	Shea CM, Edgar CM, Einhorn TA and Gerstenfeld LC: BMP treatment of C3H10T1/2 mesenchymal stem cells induces both chondrogenesis and osteogenesis. J Cell Biochem. 90:1112–1127. 2003.PubMed/NCBI View Article : Google Scholar
22	Wu N, Zhao Y, Yin Y, Zhang Y and Luo J: Identification and analysis of type II TGF-β receptors in BMP-9-induced osteogenic differentiation of C3H10T1/2 mesenchymal stem cells. Acta Biochim Biophys Sin (Shanghai). 42:699–708. 2010.PubMed/NCBI View Article : Google Scholar
23	Bruderer M, Richards RG, Alini M and Stoddart MJ: Role and regulation of RUNX2 in osteogenesis. Eur Cell Mater. 28:269–286. 2014.PubMed/NCBI View Article : Google Scholar
24	Sharma U, Pal D and Prasad R: Alkaline phosphatase: An overview. Indian J Clin Biochem. 29:269–278. 2014.PubMed/NCBI View Article : Google Scholar
25	Icer MA and Gezmen-Karadag M: The multiple functions and mechanisms of osteopontin. Clin Biochem. 59:17–24. 2018.PubMed/NCBI View Article : Google Scholar
26	Springsteen G and Wang B: Alizarin Red S as a general optical reporter for studying the binding of boronic acids with carbohydrates. Chem Commun (Camb). 17:1608–1609. 2001.PubMed/NCBI View Article : Google Scholar
27	Molchadsky A, Ezra O, Amendola PG, Krantz D, Kogan-Sakin I, Buganim Y, Rivlin N, Goldfinger N, Folgiero Y, Falcioni R, et al: p53 is required for brown adipogenic differentiation and has a protective role against diet-induced obesity. Cell Death Differ. 20:774–783. 2013.PubMed/NCBI View Article : Google Scholar
28	Tontonoz P and Spiegelman BM: Fat and beyond: The diverse biology of PPARgamma. Annu Rev Biochem. 77:289–312. 2008.PubMed/NCBI View Article : Google Scholar
29	Mehlem A, Hagberg CE, Muhl L, Eriksson U and Falkevall A: Imaging of neutral lipids by oil red O for analyzing the metabolic status in health and disease. Nat Protoc. 8:1149–1154. 2013.PubMed/NCBI View Article : Google Scholar
30	Misra UK and Pizzo SV: PFT-alpha inhibits antibody-induced activation of p53 and pro-apoptotic signaling in 1-LN prostate cancer cells. Biochem Biophys Res Commun. 391:272–276. 2010.PubMed/NCBI View Article : Google Scholar
31	Li XL, Liu YB, Ma EG, Shen WX, Li H and Zhang YN: Synergistic effect of BMP9 and TGF-β in the proliferation and differentiation of osteoblasts. Genet Mol Res. 14:7605–7615. 2015.PubMed/NCBI View Article : Google Scholar
32	Deng Y, Li L, Zhu JH, Li PP, Deng YX, Luo HH, Yang YY, He BC and Su Y: COX-2 promotes the osteogenic potential of BMP9 through TGF-β1/p38 signaling in mesenchymal stem cells. Aging (Albany NY). 13:11336–11351. 2021.PubMed/NCBI View Article : Google Scholar
33	Karkampouna S, Goumans MJ, Dijke PT, Dooley S and Julio MK: Inhibition of TGFβ type I receptor activity facilitates liver regeneration upon acute CCl4 intoxication in mice. Arch Toxicol. 90:347–357. 2016.PubMed/NCBI View Article : Google Scholar
34	Hasegawa T and Ishii M: Visualizing bone tissue in homeostatic and pathological conditions. Proc Jpn Acad Ser B Phys Biol Sci. 96:43–49. 2020.PubMed/NCBI View Article : Google Scholar
35	Manzini BM, Machado LMR, Noritomi PY and DA Silva JVL: Advances in bone tissue engineering: A fundamental review. J Biosci. 46(17)2021.PubMed/NCBI
36	Lamplot JD, Qin J, Nan G, Wang J, Liu X, Yin L, Tomal J, Li R, Shui W, Zhang H, et al: BMP9 signaling in stem cell differentiation and osteogenesis. Am J Stem Cells. 2:1–21. 2013.PubMed/NCBI
37	Wang H, Hu Y, He F, Li L, Li PP, Deng Y, Li FS, Wu K and He BC: All-trans retinoic acid and COX-2 cross-talk to regulate BMP9-induced osteogenic differentiation via Wnt/β-catenin in mesenchymal stem cells. Biomed Pharmacother. 118(109279)2019.PubMed/NCBI View Article : Google Scholar
38	Zheng W, Gu X, Sun X, Wu Q and Dan H: FAK mediates BMP9-induced osteogenic differentiation via Wnt and MAPK signaling pathway in synovial mesenchymal stem cells. Artif Cells Nanomed Biotechnol. 47:2641–2649. 2019.PubMed/NCBI View Article : Google Scholar
39	Liu Y and Gu W: p53 in ferroptosis regulation: The new weapon for the old guardian. Cell Death Differ. 29:895–910. 2022.PubMed/NCBI View Article : Google Scholar
40	Hafner A, Bulyk ML, Jambhekar A and Lahav G: The multiple mechanisms that regulate p53 activity and cell fate. Nat Rev Mol Cell Biol. 20:199–210. 2019.PubMed/NCBI View Article : Google Scholar
41	Li B, Zhang YW, Liu X, Ma L and Yang JX: Molecular mechanisms of intermuscular bone development in fish: A review. Zool Res. 42:362–376. 2021.PubMed/NCBI View Article : Google Scholar
42	Boregowda SV, Krishnappa V, Strivelli J, Haga CL, Booker CN and Phinney DG: Basal p53 expression is indispensable for mesenchymal stem cell integrity. Cell Death Differ. 25:679–692. 2018.PubMed/NCBI View Article : Google Scholar
43	Yue R, Zhou B, Shimada IS, Zhao Z and Morrison SJ: Leptin receptor promotes adipogenesis and reduces osteogenesis by regulating mesenchymal stromal cells in adult bone marrow. Cell Stem Cell. 18:782–796. 2016.PubMed/NCBI View Article : Google Scholar
44	Rosen ED and MacDougald OA: Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol. 7:885–896. 2006.PubMed/NCBI View Article : Google Scholar
45	Trivedi T, Pagnotti GM, Guise TA and Mohammad KS: The role of TGF-β in bone metastases. Biomolecules. 11(1643)2021.PubMed/NCBI View Article : Google Scholar
46	Zou ML, Chen ZH, Teng YY, Liu SY, Jia Y, Zhang KW, Sun ZL, Wu JJ, Yuan ZD, Feng Y, et al: The smad dependent TGF-β and BMP signaling pathway in bone remodeling and therapies. Front Mol Biosci. 8(593310)2021.PubMed/NCBI View Article : Google Scholar
47	Tang N, Song WX, Luo J, Luo X, Chen J, Sharff KA, Bi Y, He BC, Huang JY, Zhu GH, et al: BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/β-catenin signalling. J Cell Mol Med. 13:2448–2464. 2009.PubMed/NCBI View Article : Google Scholar
48	Huang E, Bi Y, Jiang W, Luo X, Yang K, Gao JL, Gao Y, Luo Q, Shi Q, Kim SH, et al: Conditionally immortalized mouse embryonic fibroblasts retain proliferative activity without compromising multipotent differentiation potential. PLoS One. 7(e32428)2012.PubMed/NCBI View Article : Google Scholar