SP1/TGF‑β1/SMAD2 pathway is involved in angiogenesis during osteogenesis

Ding,Ao; Bian,Ying‑Ying; Zhang,Zhi‑Hong

doi:10.3892/mmr.2020.10965

SP1/TGF‑β1/SMAD2 pathway is involved in angiogenesis during osteogenesis

Authors:
- Ao Ding
- Ying‑Ying Bian
- Zhi‑Hong Zhang
View Affiliations

Affiliations: Department of Stomatology, The First Affiliated Hospital of USTC (Anhui Provincial Hospital), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, P.R. China
Published online on: January 27, 2020 https://doi.org/10.3892/mmr.2020.10965
Pages: 1581-1589
Copyright: © Ding 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

The relationship between osteoblasts and angiogenesis is vital for bone regeneration, especially mandibular and maxillary bones. Transforming growth factor β1 (TGF‑β1) and vascular endothelial growth factor (VEGF) are closely related to angiogenesis; however, the regulatory mechanism between them remains unknown. The present study aimed to reveal this mechanism to provide novel insight for development of potential therapeutic opportunities. Western blotting and reverse transcription‑quantitative PCR was used to assess the protein and mRNA expression levels in MC3T3‑E1 preosteoblast cells and HUVECs, ELISAs were used to detect the expression levels of secreted VEGF, MTT assays were used to assess the viability of the cells, migratory ability was assessed using Transwell assays, angiogenesis assays were used to analyze the formation of blood vessels, and TGF‑β1 regulation was confirmed using a dual‑luciferase reporter assay. The overexpression of specificity protein 1 (SP1) or TGF‑β1 increased VEGF expression levels and secretion, and promoted angiogenesis of co‑cultured HUVECs. SP1 also promoted SMAD2 phosphorylation. These effects of SP1 were all reversed by the TGF‑β1 inhibitor. The VEGF inhibitor bevacizumab also reduced the SP1/TGF‑β1/SMAD2 pathway‑induced angiogenesis of preosteoblasts. In conclusion, it was demonstrated that SP1 promoted TGF‑β1 expression, activated the SMAD2 pathway and induced VEGF secretion, which may enhance angiogenic processes in preosteoblasts.

View Figures

View References

1	Okoturo E, Ogunbanjo OV and Arotiba GT: Spontaneous regeneration of the mandible: An institutional audit of regenerated bone and osteocompetent periosteum. J Oral Maxillofac Surg. 74:1660–1667. 2016. View Article : Google Scholar : PubMed/NCBI
2	Gornitsky J, Azzi AJ and Cugno S: Spontaneous osteogenesis of a traumatic mandibular defect in the pediatric population. J Craniofac Surg. 30:1999–2000. 2019. View Article : Google Scholar : PubMed/NCBI
3	Grosso A, Burger MG, Lunger A, Schaefer DJ, Banfi A and Di Maggio N: It takes two to tango: Coupling of angiogenesis and osteogenesis for bone regeneration. Front Bioeng Biotechnol. 5:682017. View Article : Google Scholar : PubMed/NCBI
4	Dickson K, Katzman S, Delgado E and Contreras D: Delayed unions and nonunions of open tibial fractures. Correlation with arteriography results. Clin Orthop Relat Res. 302:189–193. 1994.
5	Hankenson KD, Dishowitz M, Gray C and Schenker M: Angiogenesis in bone regeneration. Injury. 42:556–561. 2011. View Article : Google Scholar : PubMed/NCBI
6	Ramasamy SK, Kusumbe AP, Itkin T, Gur-Cohen S, Lapidot T and Adams RH: Regulation of hematopoiesis and osteogenesis by blood vessel-derived signals. Annu Rev Cell Dev Biol. 32:649–675. 2016. View Article : Google Scholar : PubMed/NCBI
7	Frohlich LF: Micrornas at the interface between osteogenesis and angiogenesis as targets for bone regeneration. Cells. 8:E1212019. View Article : Google Scholar : PubMed/NCBI
8	Huang B, Wang W, Li Q, Wang Z, Yan B, Zhang Z, Wang L, Huang M, Jia C, Lu J, et al: Osteoblasts secrete Cxcl9 to regulate angiogenesis in bone. Nat Commun. 7:138852016. View Article : Google Scholar : PubMed/NCBI
9	Qu J, Lu D, Guo H, Miao W, Wu G and Zhou M: MicroRNA-9 regulates osteoblast differentiation and angiogenesis via the AMPK signaling pathway. Mol Cell Biochem. 411:23–33. 2016. View Article : Google Scholar : PubMed/NCBI
10	Chen CY, Su CM, Hsu CJ, Huang CC, Wang SW, Liu SC, Chen WC, Fuh LJ and Tang CH: CCN1 promotes VEGF production in osteoblasts and induces endothelial progenitor cell angiogenesis by inhibiting miR-126 expression in rheumatoid arthritis. J Bone Miner Res. 32:34–45. 2017. View Article : Google Scholar : PubMed/NCBI
11	Jarad M, Kuczynski EA, Morrison J, Viloria-Petit AM and Coomber BL: Release of endothelial cell associated VEGFR2 during TGF-β modulated angiogenesis in vitro. BMC Cell Biol. 18:102017. View Article : Google Scholar : PubMed/NCBI
12	Wang X, Abraham S, McKenzie JAG, Jeffs N, Swire M, Tripathi VB, Luhmann UFO, Lange CAK, Zhai Z, Arthur HM, et al: LRG1 promotes angiogenesis by modulating endothelial TGF-β signalling. Nature. 499:306–311. 2013. View Article : Google Scholar : PubMed/NCBI
13	Muppala S, Xiao R, Krukovets I, Verbovetsky D, Yendamuri R, Habib N, Raman P, Plow E and Stenina-Adognravi O: Thrombospondin-4 mediates TGF-β -induced angiogenesis. Oncogene. 36:5189–5198. 2017. View Article : Google Scholar : PubMed/NCBI
14	Zhang Z, Zhang X, Zhao D, Liu B, Wang B, Yu W, Li J, Yu X, Cao F, Zheng G, et al: TGF -β1 promotes the osteoinduction of human osteoblasts via the PI3K/AKT/mTOR/S6K1 signalling pathway. Mol Med Rep. 19:3505–3518. 2019.PubMed/NCBI
15	Asparuhova MB, Caballé-Serrano J, Buser D and Chappuis V: Bone-Conditioned medium contributes to initiation and progression of osteogenesis by exhibiting synergistic TGF-β1/BMP-2 activity. Int J Oral Sci. 10:202018. View Article : Google Scholar : PubMed/NCBI
16	Wang R, Xu B and Xu H: TGF-β1 promoted chondrocyte proliferation by regulating Sp1 through MSC-exosomes derived miR-135b. Cell Cycle. 11:172018.
17	Zhang HW, Wang EW, Li LX, Yi SH, Li LC, Xu FL, Wang DL, Wu YZ and Nian WQ: A regulatory loop involving miR-29c and Sp1 elevates the TGF-β 1 mediated epithelial-to-mesenchymal transition in lung cancer. Oncotarget. 7:85905–85916. 2016.PubMed/NCBI
18	Hu J, Shan Z, Hu K, Ren F, Zhang W, Han M, Li Y, Feng K, Lei L and Feng Y: MiRNA-223 inhibits epithelial-mesenchymal transition in gastric carcinoma cells via Sp1. Int J Oncol. 49:325–335. 2016. View Article : Google Scholar : PubMed/NCBI
19	Yu S, Yerges-Armstrong LM, Chu Y, Zmuda JM and Zhang Y: Transcriptional regulation of frizzled-1 in human osteoblasts by Sp1. PLoS One. 11:e01632772016. View Article : Google Scholar : PubMed/NCBI
20	Duttenhoefer F, Biswas SK, Igwe JC, Sauerbier S and Bierhaus A: Sp1-Dependent regulation of PPARα in bone metabolism. Int J Oral Maxillofac Implants. 29:e107–e116. 2014. View Article : Google Scholar : PubMed/NCBI
21	Kim J, Lee HW, Rhee DK, Paton JC and Pyo S: Pneumolysin-induced autophagy contributes to inhibition of osteoblast differentiation through downregulation of Sp1 in human osteosarcoma cells. Biochim Biophys Acta Gen Subj. 1861:2663–2673. 2017. View Article : Google Scholar : PubMed/NCBI
22	Su F, Geng J, Li X, Qiao C, Luo L, Feng J, Dong X and Lv M: Bsp1 promotes tumor angiogenesis and invasion by activating VEGF expression in an acquired trastuzumabresistant ovarian cancer model. Oncol Rep. 38:2677–2684. 2017. View Article : Google Scholar : PubMed/NCBI
23	Hu H, Han T, Zhuo M, Wu LL, Yuan C, Wu L, Lei W, Jiao F and Wang LW: Elevated COX-2 expression promotes angiogenesis through EGFR/p38-MAPK/Sp1-dependent signalling in pancreatic cancer. Sci Rep. 7:4702017. View Article : Google Scholar : PubMed/NCBI
24	Liu M, Fan F, Shi P, Tu M, Yu C, Yu C and Du M: Lactoferrin promotes MC3T3-E1 osteoblast cells proliferation via MAPK signaling pathways. Int J Biol Macromol. 107:137–143. 2018. View Article : Google Scholar : PubMed/NCBI
25	Zhai F, Song N, Ma J, Gong W, Tian H, Li X, Jiang C and Wang H: FGF18 inhibits MC3T3E1 cell osteogenic differentiation via the ERK signaling pathway. Mol Med Rep. 16:4127–4132. 2017. View Article : Google Scholar : PubMed/NCBI
26	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
27	Chen Y, Zhang Q, Charles EB, Wu Y, Zhang L, Dong J and Han Y: VEGF overespression promotes the proliferation and differentiation of human adipose-derived stem cells. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 33:352–356. 2017.(In Chinese). PubMed/NCBI
28	Bhattacharya R, Fan F, Wang R, Ye X, Xia L, Boulbes D and Ellis LM: Intracrine VEGF signalling mediates colorectal cancer cell migration and invasion. Br J Cancer. 117:848–855. 2017. View Article : Google Scholar : PubMed/NCBI