Casein kinase 2-interaction protein-1 regulates the osteogenic ability of bone marrow mesenchymal stem cells in osteoporosis rats

doi:10.12307/2023.249

Abstract

Abstract: BACKGROUND: At present, in vitro studies on the molecular mechanism of casein kinase 2-interaction protein-1 (CKIP-1) mainly focus on gene knockout mouse-derived osteoblasts or bone marrow mesenchymal stem cells. There are few reports focusing on the expression of CKIP-1 in bone marrow mesenchymal stem cells derived from osteoporosis model rats.
OBJECTIVE: To investigate the changes in osteogenic differentiation of bone marrow mesenchymal stem cells before and after down-regulation of CKIP-1 gene.
METHODS: An osteoporotic rat model was induced by retinoic acid gavage. The bone marrow mesenchymal stem cells of the osteoporosis group and the normal group were cultured in vitro by the whole bone marrow adherence method. After osteogenic induction, Alizarin red staining, alkaline phosphatase staining and real-time quantitative RT-PCR were utilized to detect the relative expression of osteopontin and Runx2 mRNA. The dynamic expression of CKIP-1 in the two groups of cells was detected by using real-time quantitative RT-PCR. CKIP-1 gene was silenced by gene transfection. After osteogenic induction, alizarin red staining, alkaline phosphatase staining, and real-time quantitative RT-PCR were performed to detect the relative expression of osteopontin and Runx2 mRNA.
RESULTS AND CONCLUSION: (1) Compared with the normal group, alizarin red staining of calcium nodules, alkaline phosphatase activity and mRNA levels of osteopontin and Runx2 in bone marrow mesenchymal stem cells decreased in the osteoporosis group (P < 0.05). The dynamic expression level of CKIP-1 gene in bone marrow mesenchymal stem cells was generally higher in the osteoporosis group. (2) Compared with bone marrow mesenchymal stem cells of osteoporosis group without down-regulation of CKIP-1, after down-regulation of CKIP-1 gene expression, the quantification of calcium nodules and alkaline phosphatase activity by alizarin red staining, mRNA levels of osteopontin and Runx2 were significantly increased in bone marrow mesenchymal stem cells of the osteoporosis group (P < 0.05). (3) Therefore, osteogenic ability of bone marrow mesenchymal stem cells in rats with retinoic acid-induced osteoporosis is reduced and down-regulation of CKIP-1 gene can partially improve their osteogenic differentiation ability.

Key words: rat, retinoic acid, CKIP-1, osteoporosis, bone marrow mesenchymal stem cell, osteogenic differentiation

CLC Number:

Long Yanming, Xie Mengsheng, Huang Jiajie, Xue Wenli, Rong Hui, Li Xiaojie. Casein kinase 2-interaction protein-1 regulates the osteogenic ability of bone marrow mesenchymal stem cells in osteoporosis rats[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(6): 878-882.

Figures/Tables 3

References

[1] JONASSON G, RYTHÉN M. Alveolar bone loss in osteoporosis: a loaded and cellular affair. Clin Cosmet Investig Dent. 2016;8:95-103.
[2] LI X, CHEN M, LI L, et al. Extracorporeal shock wave therapy: a potential adjuvant treatment for peri-implantitis. Med Hypotheses. 2010;74(1): 120-122.
[3] BOSC DG, GRAHAM KC, SAULNIER RB, et al. Identification and characterization of CKIP-1, a novel pleckstrin homology domain-containing protein that interacts with protein kinase CK2. J Biol Chem. 2000;275(19):14295-14306.
[4] LU K, YIN X, WENG T, et al. Targeting WW domains linker of HECT-type ubiquitin ligase Smurf1 for activation by CKIP-1. Nat Cell Biol. 2008; 10(8):994-1002.
[5] 龙燕鸣,谢梦生,薛文利,等.维甲酸诱导骨质疏松模型大鼠骨髓间充质干细胞的特征[J].中国组织工程研究,2019,23(17):2637-2643.
[6] HOLLICK RJ, REID DM. Role of bisphosphonates in the management of postmenopausal osteoporosis: an update on recent safety anxieties. Menopause Int. 2011;17(2):66-72.
[7] SCHILCHER J, MICHAËLSSON K, Aspenberg P. Bisphosphonate use and atypical fractures of the femoral shaft. N Engl J Med. 2011;364(18): 1728-1737.
[8] WANG Y, NISHIDA S, BOUDIGNON BM, et al. IGF-I receptor is required for the anabolic actions of parathyroid hormone on bone. J Bone Miner Res. 2007;22(9):1329-1337.
[9] BIKLE DD, SAKATA T, LEARY C, et al. Insulin-like growth factor I is required for the anabolic actions of parathyroid hormone on mouse bone. J Bone Miner Res. 2002;17(9):1570-1578.
[10] 韩辰,谢幼专,袁君杰.小干扰RNA技术与骨质疏松症治疗[J].国际骨科学杂志,2014,35(2):86-88.
[11] HOWARD KA, PALUDAN SR, BEHLKE MA, et al. Chitosan/siRNA nanoparticle-mediated TNF-alpha knockdown in peritoneal macrophages for anti-inflammatory treatment in a murine arthritis model. Mol Ther. 2009;17(1):162-168.
[12] KRAMER PR, PURI J, BELLINGER LL. Knockdown of Fcγ receptor III in an arthritic temporomandibular joint reduces the nociceptive response in rats. Arthritis Rheum. 2010;62(10):3109-3118.
[13] WANG Y, GRAINGER DW. RNA therapeutics targeting osteoclast-mediated excessive bone resorption. Adv Drug Deliv Rev. 2012;64(12): 1341-1357.
[14] BUCKBINDER L, CRAWFORD DT, QI H, et al. Proline-rich tyrosine kinase 2 regulates osteoprogenitor cells and bone formation, and offers an anabolic treatment approach for osteoporosis. Proc Natl Acad Sci U S A. 2007;104(25):10619-10624.
[15] LIU J, LU C, WU X, et al. Targeting osteoblastic casein kinase-2 interacting protein-1 to enhance Smad-dependent BMP signaling and reverse bone formation reduction in glucocorticoid-induced osteoporosis. Sci Rep. 2017;7:41295.
[16] MCKEE MD, PEDRAZA CE, KAARTINEN MT. Osteopontin and wound healing in bone. Cells Tissues Organs. 2011;194(2-4):313-319.
[17] NIE J, LIU L, HE F, et al. CKIP-1: a scaffold protein and potential therapeutic target integrating multiple signaling pathways and physiological functions. Ageing Res Rev. 2013;12(1):276-281.
[18] ZHANG G, GUO B, WU H, et al. A delivery system targeting bone formation surfaces to facilitate RNAi-based anabolic therapy. Nat Med. 2012;18(2):307-314.
[19] ZHANG L, WU K, SONG W, et al. Chitosan/siCkip-1 biofunctionalized titanium implant for improved osseointegration in the osteoporotic condition. Sci Rep. 2015;5:10860.
[20] ZHOU ZC, CHE L, KONG L, et al. CKIP-1 silencing promotes new bone formation in rat mandibular distraction osteogenesis. Oral Surg Oral Med Oral Pathol Oral Radiol. 2017;123(1):e1-e9.
[21] BUSILLO JM, BENOVIC JL. Regulation of CXCR4 signaling. Biochim Biophys Acta. 2007;1768(4):952-963.
[22] GUO B, ZHANG B, ZHENG L, et al. Therapeutic RNA interference targeting CKIP-1 with a cross-species sequence to stimulate bone formation. Bone. 2014;59:76-88.
[23] XU P, GUO X, ZHANG YG, et al. The effect of retinoic acid on induction of osteoporotic model rats and the possible mechanism. Sichuan Da Xue Xue Bao Yi Xue Ban. 2005;36(2):229-232.
[24] LIU Q, ZHANG X, JIAO Y, et al. In vitro cell behaviors of bone mesenchymal stem cells derived from normal and postmenopausal osteoporotic rats. Int J Mol Med. 2018;41(2):669-678.
[25] PINO AM, ROSEN CJ, RODRÍGUEZ JP. In osteoporosis, differentiation of mesenchymal stem cells (MSCs) improves bone marrow adipogenesis. Biol Res. 2012;45(3):279-287.