Ckip‑1 regulates C3H10T1/2 mesenchymal cell proliferation and osteogenic differentiation via Lrp5

Huang,Xin; Liang,Jianfei; Gao,Ye; Hou,Yan; Song,Yu; Kong,Liang

doi:10.3892/etm.2021.9773

Ckip‑1 regulates C3H10T1/2 mesenchymal cell proliferation and osteogenic differentiation via Lrp5

Authors:
- Xin Huang
- Jianfei Liang
- Ye Gao
- Yan Hou
- Yu Song
- Liang Kong
View Affiliations

Affiliations: State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Orthodontics, Qingdao Stomatological Hospital, Qingdao, Shandong 266001, P.R. China
Published online on: February 10, 2021 https://doi.org/10.3892/etm.2021.9773
Article Number: 342
Copyright: © Huang 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

Casein kinase‑2 interaction protein‑1 (Ckip‑1) is a negative regulator of bone formation. The identification of novel Ckip‑1‑related targets and their associated signaling pathways that regulate mesenchymal stem cell (MSC) osteogenic differentiation is required. The present study aimed to evaluate the effects of Ckip‑1 knockdown on C3H10T1/2 MSC proliferation and osteogenic differentiation, and to explore the role of the canonical Wnt‑signaling receptor Lrp5. Ckip‑1‑knockdown (shCkip‑1), Ckip‑1‑overexpression (Ckip‑1) and their corresponding control [shCtrl and empty vector (EV), respectively] cell groups were used in the present study. Immunofluorescence localization of Ckip‑1 was observed. The expression of the key molecules of the canonical Wnt signaling pathway was examined in C3H10T1/2 cells following osteogenic induction. Moreover, the effects of Lrp5 knockdown in the presence or absence of Ckip‑1 knockdown were examined on C3H10T1/2 cell proliferation and osteogenic differentiation. The results indicated an increase in cell proliferation and osteogenic differentiation in the shCkip‑1 group compared with the shCtrl group. The expression levels of LDL receptor related protein 5 (Lrp5), lymphoid enhancer binding factor 1 (Lef1) and transcription factor 1 in C3H10T1/2 cells were significantly increased in shCkip‑1 cells following 7‑day osteoinduction compared with shCtrl cells. Moreover, the involvement of Lrp5 in shCkip‑1‑induced osteogenic differentiation of C3H10T1/2 cells was further verified. The results indicated that Ckip‑1 reduced C3H10T1/2 MSC proliferation and osteogenic differentiation via the canonical Wnt‑signaling receptor Lrp5, which is essential for the improvement of bone tissue engineering.

View Figures

View References

1	Li L, Lu H, Zhao Y, Luo J, Luo J, Yang L, Liu W and He Q: Functionalized cell-free scaffolds for bone defect repair inspired by self-healing of bone fractures: A review and new perspectives. Mater Sci Eng C Mater Biol Appl. 98:1241–1251. 2019.PubMed/NCBI View Article : Google Scholar
2	Liu M and Lv Y: Reconstructing bone with natural bone graft: A review of in vivo studies in bone defect animal model. Nanomaterials (Basel). 8(E999)2018.PubMed/NCBI View Article : Google Scholar
3	Wei X, Liu B, Liu G, Yang F, Cao F, Dou X, Yu W, Wang B, Zheng G, Cheng L, et al: Mesenchymal stem cell-loaded porous tantalum integrated with biomimetic 3D collagen-based scaffold to repair large osteochondral defects in goats. Stem Cell Res Ther. 10(72)2019.PubMed/NCBI View Article : Google Scholar
4	Chen X, Fan H, Deng X, Wu L, Yi T, Gu L, Zhou C, Fan Y and Zhang X: Scaffold structural microenvironmental cues to guide tissue regeneration in bone tissue applications. Nanomaterials (Basel). 8(E960)2018.PubMed/NCBI View Article : Google Scholar
5	White KA and Olabisi RM: Spatiotemporal control strategies for bone formation through tissue engineering and regenerative medicine approaches. Adv Healthc Mater. 8(e1801044)2019.PubMed/NCBI View Article : Google Scholar
6	El-Rashidy AA, Roether JA, Harhaus L, Kneser U and Boccaccini AR: Regenerating bone with bioactive glass scaffolds: A review of in vivo studies in bone defect models. Acta Biomater. 62:1–28. 2017.PubMed/NCBI View Article : Google Scholar
7	Leyendecker Junior A, Gomes Pinheiro CC, Lazzaretti Fernandes T and Franco Bueno D: The use of human dental pulp stem cells for in vivo bone tissue engineering: A systematic review. J Tissue Eng. 9(2041731417752766)2018.PubMed/NCBI View Article : Google Scholar
8	Wehner D and Weidinger G: Signaling networks organizing regenerative growth of the zebrafish fin. Trends Genet. 31:336–343. 2015.PubMed/NCBI View Article : Google Scholar
9	Ren L, Chen H, Song J, Chen X, Lin C, Zhang X, Hou N, Pan J, Zhou Z, Wang L, et al: MiR-454-3p-mediated Wnt/β-catenin signaling antagonists suppression promotes breast cancer metastasis. Theranostics. 9:449–465. 2019.PubMed/NCBI View Article : Google Scholar
10	Yuan K, Shamskhou EA, Orcholski ME, Nathan A, Reddy S, Honda H, Mani V, Zeng Y, Ozen MO, Wang L, et al: Loss of endothelial derived WNT5A is associated with reduced pericyte recruitment and small vessel loss in pulmonary arterial hypertension. Circulation. 139:1710–1724. 2019.PubMed/NCBI View Article : Google Scholar
11	Gong B, Shen W, Xiao W, Meng Y, Meng A and Jia S: The Sec14-like phosphatidylinositol transfer proteins Sec14l3/SEC14L2 act as GTPase proteins to mediate Wnt/Ca²⁺ signaling. Elife. 6(e26362)2017.PubMed/NCBI View Article : Google Scholar
12	Baron R and Kneissel M: WNT signaling in bone homeostasis and disease: From human mutations to treatments. Nat Med. 19:179–192. 2013.PubMed/NCBI View Article : Google Scholar
13	Taipaleenmäki H, Abdallah BM, Aldahmash A, Säämänen AM and Kassem M: Wnt signalling mediates the cross-talk between bone marrow derived pre-adipocytic and pre-osteoblastic cell populations. Exp Cell Res. 317:745–756. 2011.PubMed/NCBI View Article : Google Scholar
14	Day TF, Guo X, Garrett-Beal L and Yang Y: Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell. 8:739–750. 2005.PubMed/NCBI View Article : Google Scholar
15	D'Alimonte I, Lannutti A, Pipino C, Di Tomo P, Pierdomenico L, Cianci E, Antonucci I, Marchisio M, Romano M, Stuppia L, et al: Wnt signaling behaves as a ‘master regulator’ in the osteogenic and adipogenic commitment of human amniotic fluid mesenchymal stem cells. Stem Cell Rev Rep. 9:642–654. 2013.PubMed/NCBI View Article : Google Scholar
16	Zuo R, Liu M, Wang Y, Li J, Wang W, Wu J, Sun C, Li B, Wang Z, Lan W, et al: BM-MSC-derived exosomes alleviate radiation-induced bone loss by restoring the function of recipient BM-MSCs and activating Wnt/β-catenin signaling. Stem Cell Res Ther. 10(30)2019.PubMed/NCBI View Article : Google Scholar
17	Fan J, An X, Yang Y, Xu H, Fan L, Deng L, Li T, Weng X, Zhang J and Zhao RC: MiR-1292 targets FZD4 to regulate senescence and osteogenic differentiation of stem cells in TE/SJ/mesenchymal tissue system via the Wnt/β-catenin pathway. Aging Dis. 9:1103–1121. 2018.PubMed/NCBI View Article : Google Scholar
18	Williams BO: LRP5: From bedside to bench to bone. Bone. 102:26–30. 2017.PubMed/NCBI View Article : Google Scholar
19	Li T, Li H, Wang Y, Li T, Fan J, Xiao K, Zhao RC and Weng X: microRNA-23a inhibits osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting LRP5. Int J Biochem Cell Biol. 72:55–62. 2016.PubMed/NCBI View Article : Google Scholar
20	Borrell-Pagès M, Romero JC and Badimon L: LRP5 deficiency down-regulates Wnt signalling and promotes aortic lipid infiltration in hypercholesterolaemic mice. J Cell Mol Med. 19:770–777. 2015.PubMed/NCBI View Article : Google Scholar
21	Semenov MV and He X: LRP5 mutations linked to high bone mass diseases cause reduced LRP5 binding and inhibition by SOST. J Biol Chem. 281:38276–38284. 2006.PubMed/NCBI View Article : Google Scholar
22	Nie J, Liu L, He F, Fu X, Han W and Zhang L: CKIP-1: A scaffold protein and potential therapeutic target integrating multiple signaling pathways and physiological functions. Ageing Res Rev. 12:276–281. 2013.PubMed/NCBI View Article : Google Scholar
23	Lu K, Yin X, Weng T, Xi S, Li L, Xing G, Cheng X, Yang X, Zhang L and He F: Targeting WW domains linker of HECT-type ubiquitin ligase Smurf1 for activation by CKIP-1. Nat Cell Biol. 10:994–1002. 2008.PubMed/NCBI View Article : Google Scholar
24	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.PubMed/NCBI View Article : Google Scholar
25	Chen BY, Wang X, Chen LW and Luo ZJ: Molecular targeting regulation of proliferation and differentiation of the bone marrow-derived mesenchymal stem cells or mesenchymal stromal cells. Curr Drug Targets. 13:561–571. 2012.PubMed/NCBI View Article : Google Scholar
26	Lee DJ, Kwon J, Current L, Yoon K, Zalal R, Hu X, Xue P and Ko CC: Osteogenic potential of mesenchymal stem cells from rat mandible to regenerate critical sized calvarial defect. J Tissue Eng. 10(2041731419830427)2019.PubMed/NCBI View Article : Google Scholar
27	Moore ER and Jacobs CR: The primary cilium as a signaling nexus for growth plate function and subsequent skeletal development. J Orthop Res. 36:533–545. 2018.PubMed/NCBI View Article : Google Scholar
28	Reznikoff CA, Brankow DW and Heidelberger C: Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 33:3231–3238. 1973.PubMed/NCBI
29	Hashimi SM: Exogenous noggin binds the BMP-2 receptor and induces alkaline phosphatase activity in osteoblasts. J Cell Biochem. 120:13237–13242. 2019.PubMed/NCBI View Article : Google Scholar
30	Li D, Zhu H, Liang C, Li W, Xing G, Ma L, Ding L, Zhang Y, He F and Zhang L: CKIP-1 suppresses the adipogenesis of mesenchymal stem cells by enhancing HDAC1-associated repression of C/EBPα. J Mol Cell Biol. 6:368–379. 2014.PubMed/NCBI View Article : Google Scholar
31	Safi A, Vandromme M, Caussanel S, Valdacci L, Baas D, Vidal M, Brun G, Schaeffer L and Goillot E: Role for the pleckstrin homology domain-containing protein CKIP-1 in phosphatidylinositol 3-kinase-regulated muscle differentiation. Mol Cell Biol. 24:1245–1255. 2004.PubMed/NCBI View Article : Google Scholar
32	Zhang L, Xing G, Tie Y, Tang Y, Tian C, Li L, Sun L, Wei H, Zhu Y and He F: Role for the pleckstrin homology domain-containing protein CKIP-1 in AP-1 regulation and apoptosis. EMBO J. 24:766–778. 2005.PubMed/NCBI View Article : Google Scholar
33	Bosc DG, Graham KC, Saulnier RB, Zhang C, Prober D, Gietz RD and Litchfield DW: Identification and characterization of CKIP-1, a novel pleckstrin homology domain-containing protein that interacts with protein kinase CK2. J Biol Chem. 275:14295–14306. 2000.PubMed/NCBI View Article : Google Scholar
34	Kato M, Patel MS, Levasseur R, Lobov I, Chang BHJ, Glass DA II, Hartmann C, Li L, Hwang TH, Brayton CF, et al: Cbfa1-independent decrease in osteoblast proliferation, osteopenia, and persistent embryonic eye vascularization in mice deficient in Lrp5, a Wnt coreceptor. J Cell Biol. 157:303–314. 2002.PubMed/NCBI View Article : Google Scholar
35	Cui Y, Niziolek PJ, MacDonald BT, Zylstra CR, Alenina N, Robinson DR, Zhong Z, Matthes S, Jacobsen CM, Conlon RA, et al: Lrp5 functions in bone to regulate bone mass. Nat Med. 17:684–691. 2011.PubMed/NCBI View Article : Google Scholar
36	Hu H, Hilton MJ, Tu X, Yu K, Ornitz DM and Long F: Sequential roles of Hedgehog and Wnt signaling in osteoblast development. Development. 132:49–60. 2005.PubMed/NCBI View Article : Google Scholar
37	Hill TP, Später D, Taketo MM, Birchmeier W and Hartmann C: Canonical Wnt/beta-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell. 8:727–738. 2005.PubMed/NCBI View Article : Google Scholar