Mechanical strain promotes osteoblastic differentiation through integrin-β1-mediated β-catenin signaling

Yan,Yuxian; Sun,Haoyang; Gong,Yuanwei; Yan,Zhixiong; Zhang,Xizheng; Guo,Yong; Wang,Yang

doi:10.3892/ijmm.2016.2636

Mechanical strain promotes osteoblastic differentiation through integrin-β1-mediated β-catenin signaling

Authors:
- Yuxian Yan
- Haoyang Sun
- Yuanwei Gong
- Zhixiong Yan
- Xizheng Zhang
- Yong Guo
- Yang Wang
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Affiliations: College of Biotechnology, Guilin Medical University, Guilin, Guangxi 541100, P.R. China, School of Physical Science and Technology, Lanzhou University, Lanzhou, Gansu 730000, P.R. China
Published online on: June 10, 2016 https://doi.org/10.3892/ijmm.2016.2636
Pages: 594-600

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Abstract

As integrins are mechanoresponsive, there exists an intimate relationship between integrins and mechanical strain. Integrin-β1 mediates the impact of mechanical strain on bone. Mechanical strain induces bone formation through the activation of β-catenin pathways, which suggests that integrin-β1 mediates β-catenin signaling in osteoblasts in response to mechanical strain. In the present study, we examined the role of integrin-β1 in Wnt/β-catenin signal transduction in mechanically strained osteoblasts. MC3T3-E1 osteoblastic cells were transfected with integrin-β1 small interfering RNA (si-Itgβ1), and exposed to mechanical tensile strain of 2,500 microstrain (µε) using a four-point bending device. The mechanical strain enhanced the mRNA expression of integrin-β1, the protein levels of phosphorylated (p-) glycogen synthase kinase-3β (GSK‑3β) and β-catenin, simultaneously increased the mRNA levels of runt-related transcriptional factor 2 (Runx2) and osteocalcin (OCN), the protein levels of bone morphogenetic protein (BMP)-2 and -4 and enhanced the alkaline phosphatase (ALP) activity of the ME3T3-E1 cells. The elevations were inhibited by si-Itgβ1. Additionally, the mechanical strain induced the nuclear translocation of β-catenin into the nucleus, which was also inhibited by si-Itgβ1. These findings indicated that mechanical strain promoted osteoblastic differentiation through integrin‑β1‑mediated β-catenin signaling.

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1	Pommerenke H, Schmidt C, Dürr F, Nebe B, Lüthen F, Muller P and Rychly J: The mode of mechanical integrin stressing controls intracellular signaling in osteoblasts. J Bone Miner Res. 17:603–611. 2002. View Article : Google Scholar : PubMed/NCBI
2	Shyy JY and Chien S: Role of integrins in endothelial mechanosensing of shear stress. Circ Res. 91:769–775. 2002. View Article : Google Scholar : PubMed/NCBI
3	Gohel AR, Hand AR and Gronowicz GA: Immunogold localization of beta 1-integrin in bone: effect of glucocorticoids and insulin-like growth factor I on integrins and osteocyte formation. J Histochem Cytochem. 43:1085–1096. 1995. View Article : Google Scholar : PubMed/NCBI
4	Carvalho RS, Scott JE and Yen EH: The effects of mechanical stimulation on the distribution of beta 1 integrin and expression of beta 1-integrin mRNA in TE-85 human osteosarcoma cells. Arch Oral Biol. 40:257–264. 1995. View Article : Google Scholar : PubMed/NCBI
5	Zimmerman D, Jin F, Leboy P, Hardy S and Damsky C: Impaired bone formation in transgenic mice resulting from altered integrin function in osteoblasts. Dev Biol. 220:2–15. 2000. View Article : Google Scholar : PubMed/NCBI
6	Kapur S, Baylink DJ and Lau KH: Fluid flow shear stress stimulates human osteoblast proliferation and differentiation through multiple interacting and competing signal transduction pathways. Bone. 32:241–251. 2003. View Article : Google Scholar : PubMed/NCBI
7	Yeh CR, Chiu JJ, Lee CI, Lee PL, Shih YT, Sun JS, Chien S and Cheng CK: Estrogen augments shear stress-induced signaling and gene expression in osteoblast-like cells via estrogen receptor-mediated expression of beta1-integrin. J Bone Miner Res. 25:627–639. 2010. View Article : Google Scholar
8	Yan YX, Gong YW, Guo Y, Lv Q, Guo C, Zhuang Y, Zhang Y, Li R and Zhang XZ: Mechanical strain regulates osteoblast proliferation through integrin-mediated ERK activation. PLoS One. 7:e357092012. View Article : Google Scholar : PubMed/NCBI
9	Zeng Q, Guo Y, Liu Y, Li R, Zhang X, Liu L, Wang Y, Zhang X and Zou X: Integrin-β1, not integrin-β5, mediates osteoblastic differentiation and ECM formation promoted by mechanical tensile strain. Biol Res. 48:252015. View Article : Google Scholar
10	Robinson JA, Chatterjee-Kishore M, Yaworsky PJ, Cullen DM, Zhao W, Li C, Kharode Y, Sauter L, Babij P, Brown EL, et al: Wnt/beta-catenin signaling is a normal physiological response to mechanical loading in bone. J Biol Chem. 281:31720–31728. 2006. View Article : Google Scholar : PubMed/NCBI
11	Case N, Ma M, Sen B, Xie Z, Gross TS and Rubin J: Beta-catenin levels influence rapid mechanical responses in osteoblasts. J Biol Chem. 283:29196–29205. 2008. View Article : Google Scholar : PubMed/NCBI
12	Norvell SM, Alvarez M, Bidwell JP and Pavalko FM: Fluid shear stress induces beta-catenin signaling in osteoblasts. Calcif Tissue Int. 75:396–404. 2004. View Article : Google Scholar : PubMed/NCBI
13	Premaraj S, Souza I and Premaraj T: Mechanical loading activates β-catenin signaling in periodontal ligament cells. Angle Orthod. 81:592–599. 2011. View Article : Google Scholar : PubMed/NCBI
14	Rallis C, Pinchin SM and Ish-Horowicz D: Cell-autonomous integrin control of Wnt and Notch signalling during somitogenesis. Development. 137:3591–3601. 2010. View Article : Google Scholar : PubMed/NCBI
15	Liu Y, Chattopadhyay N, Qin S, Szekeres C, Vasylyeva T, Mahoney ZX, Taglienti M, Bates CM, Chapman HA, Miner JH and Kreidberg JA: Coordinate integrin and c-Met signaling regulate Wnt gene expression during epithelial morphogenesis. Development. 136:843–853. 2009. View Article : Google Scholar : PubMed/NCBI
16	Kim SA, Das S, Lee H, Kim JH, Kim SJ, Song YM, Kim IS, Hwang SJ and Byun KM: A surface plasmon resonance sensor for quantitative analysis of mineralization of osteoblast cells. Proceedings of IEEE Sensors. Jan;2010. View Article : Google Scholar
17	Gan Q, Lu X, Yuan Y, Qian J, Zhou H, Lu X, Shi J and Liu C: A magnetic, reversible pH-responsive nanogated ensemble based on Fe₃O₄ nanoparticles-capped mesoporous silica. Biomaterials. 32:1932–1942. 2011. View Article : Google Scholar
18	Tang LL, Wang YL, Pan J and Cai SX: The effect of step-wise increased stretching on rat calvarial osteoblast collagen production. J Biomech. 37:157–161. 2004. View Article : Google Scholar
19	Wang L, Zhang XZ, Guo Y, Chen XZ, Li RX, Liu L, Shi CH, Guo C and Zhang Y: Involvement of BMPs/Smad signaling pathway in mechanical response in osteoblasts. Cell Physiol Biochem. 26:1093–1102. 2010. View Article : Google Scholar
20	Bottlang M, Simnacher M, Schmitt H, Brand RA and Claes L: A cell strain system for small homogeneous strain applications. Biomed Tech (Berl). 42:305–309. 1997. View Article : Google Scholar
21	Owan I, Burr DB, Turner CH, Qiu J, Tu Y, Onyia JE and Duncan RL: Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain. Am J Physiol. 273:C810–C815. 1997.PubMed/NCBI
22	Day TF, Guo X, Garrett-Beal L and Yang Y: Wnt/β-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell. 8:739–750. 2005. View Article : Google Scholar : PubMed/NCBI
23	Hill TP, Später D, Taketo MM, Birchmeier W and Hartmann C: Canonical Wnt/β-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell. 8:727–738. 2005. View Article : Google Scholar : PubMed/NCBI
24	Fang X, Yu SX, Lu Y, Bast RC Jr, Woodgett JR and Mills GB: Phosphorylation and inactivation of glycogen synthase kinase 3 by protein kinase A. Proc Natl Acad Sci USA. 97:11960–11965. 2000. View Article : Google Scholar : PubMed/NCBI
25	Beck GR Jr, Zerler B and Moran E: Phosphate is a specific signal for induction of osteopontin gene expression. Proc Natl Acad Sci USA. 97:8352–8357. 2000. View Article : Google Scholar : PubMed/NCBI
26	Wu M, Hesse E, Morvan F, Zhang JP, Correa D, Rowe GC, Kiviranta R, Neff L, Philbrick WM, Horne WC and Baron R: Zfp521 antagonizes Runx2, delays osteoblast differentiation in vitro, and promotes bone formation in vivo. Bone. 44:528–536. 2009. View Article : Google Scholar :
27	Mahalingam CD, Datta T, Patil RV, Kreider J, Bonfil RD, Kirkwood KL, Goldstein SA, Abou-Samra AB and Datta NS: Mitogen-activated protein kinase phosphatase 1 regulates bone mass, osteoblast gene expression, and responsiveness to parathyroid hormone. J Endocrinol. 211:145–156. 2011. View Article : Google Scholar : PubMed/NCBI
28	Jang WG, Kim EJ and Koh JT: Tunicamycin negatively regulates BMP2-induced osteoblast differentiation through CREBH expression in MC3T3E1 cells. BMB Rep. 44:735–740. 2011. View Article : Google Scholar : PubMed/NCBI
29	Wan M and Cao X: BMP signaling in skeletal development. Biochem Biophys Res Commun. 328:651–657. 2005. View Article : Google Scholar : PubMed/NCBI
30	Liedert A, Kaspar D, Blakytny R, Claes L and Ignatius A: Signal transduction pathways involved in mechanotransduction in bone cells. Biochem Biophys Res Commun. 349:1–5. 2006. View Article : Google Scholar : PubMed/NCBI
31	Papachroni KK, Karatzas DN, Papavassiliou KA, Basdra EK and Papavassiliou AG: Mechanotransduction in osteoblast regulation and bone disease. Trends Mol Med. 15:208–216. 2009. View Article : Google Scholar : PubMed/NCBI
32	Lau KH, Kapur S, Kesavan C and Baylink DJ: Up-regulation of the Wnt, estrogen receptor, insulin-like growth factor-I, and bone morphogenetic protein pathways in C57BL/6J osteoblasts as opposed to C3H/HeJ osteoblasts in part contributes to the differential anabolic response to fluid shear. J Biol Chem. 281:9576–9588. 2006. View Article : Google Scholar : PubMed/NCBI
33	Lal H, Verma SK, Smith M, Guleria RS, Lu G, Foster DM and Dostal DE: Stretch-induced MAP kinase activation in cardiac myocytes: differential regulation through β1-integrin and focal adhesion kinase. J Mol Cell Cardiol. 43:137–147. 2007. View Article : Google Scholar : PubMed/NCBI
34	Armstrong VJ, Muzylak M, Sunters A, Zaman G, Saxon LK, Price JS and Lanyon LE: Wnt/β-catenin signaling is a component of osteoblastic bone cell early responses to load-bearing and requires estrogen receptor alpha. J Biol Chem. 282:20715–20727. 2007. View Article : Google Scholar : PubMed/NCBI
35	Yan YX, Song M, Guo C, Guo Y, Gong YW, Li RX and Zhang XZ: The effects of substrate-streching strajn on the BMP-2 mRNA expression in three kinds of mouse cell lines. Chin J Gerontol. 30:109–112. 2010.
36	Gong YW, Yan YX, Zhang Y, Zhang XZ and Guo Y: The effect of substrate-stretching strain on the expression of Runx2 in mouse osteoblasts. Chin J Osteoporos. 17:185–188. 2011.
37	Liu L, Guo Y, Wan ZM, Shi CH, Li JY, Li RX, Hao QX, Li H and Zhang XZ: Effects of phytoestrogen a-ZAL and mechanical stimulation on proliferation, osteoblastic differentiation, and OPG/RANKL expression in MC3T3-E1 pre-osteoblasts. Cell Mol Bioeng. 5:427–439. 2012. View Article : Google Scholar
38	Aubin JE, Liu F, Malaval L and Gupta AK: Osteoblast and chondroblast differentiation. Bone. 17(Suppl 2): 77S–83S. 1995. View Article : Google Scholar : PubMed/NCBI