Int J Stomatol ›› 2019, Vol. 46 ›› Issue (4): 420-425.doi: 10.7518/gjkq.2019048

• Original Articles • Previous Articles     Next Articles

Effect of CaMKⅡδ gene silencing on osteoclast differentiation and c-fos/c-jun/CREB gene

Zhang Yuhong1,Qi Mengchun1(),Dong Wei1,Sun Hong2   

  1. 1. Dept. of Oral and Maxillofacial Surgery, College of Stomatology, North China University of Science and Technology, Tangshan 063000, China
    2. Dept. of Pathology, College of Basic Medicine, North China University of Science and Technology, Tangshan 063000, China
  • Received:2018-07-22 Revised:2019-03-01 Online:2019-07-01 Published:2019-07-12
  • Supported by:
    This study was supported by National Natural Science Foundation of China(81270965);National Natural Science Foundation of Hebei Province(20-17209114)

Abstract:

Objective This study aims to explore the effect of calmodulin-dependent kinase (CaMK) Ⅱδ gene silencing on osteoclast differentiation, investigate the roles of the c-fos, c-jun and CREB genes in CaMKⅡδ regulation of osteoclast differentiation, and reveal the molecular mechanism underlying osteoclast differentiation.Methods A CaMKⅡδ RNA interference vector was constructed and transfected into mouse RAW264.7 cells to determine interference efficiency. The cells were transfected with the virus, induced with 50 ng·mL -1 nuclear factor-κB receptor activator ligand and then harvested after 5 days. The effect of silencing the CaMKⅡδ gene on osteoclast differentiation was detected by tartrate-resistant acid phosphatase detection and bovine bone resorption lacuna. After 48 h, the expression levels of the c-fos, c-jun and CREB genes were detected by real-time polymerase chain reaction and Western blot.Results The interference efficiencies of the CaMKⅡδ RNA interference vector at the mRNA and protein levels were 70.6% and 72.2%, respectively. CaMKⅡδ RNA interference significantly reduced the mRNA levels of c-fos, c-jun and CREB by 74%, 49% and 24%, respectively. CaMKⅡδ RNA interference significantly reduced the protein levels of c-Fos, c-Jun and CREB by 74.3%, 61.3% and 59.2%, respectively.Conclusion CaMKⅡδ RNA interference can significantly inhibit the expression of c-fos, c-jun and CREB at the mRNA and protein levels. These results indicate that c-Fos, c-Jun and CREB are involved in the regulatory effects of CaMKⅡδ on osteoclast differentiation.

Key words: osteoclast, calmodulin-dependent kinase Ⅱδ, c-Fos, c-Jun, cyclic adenosine monophosphate responsive element-binding protein

CLC Number: 

  • Q786

TrendMD: 

Fig 1

CaMKⅡδ mRNA levels detected by real-time PCR"

Fig 2

CaMKⅡδ levels detected by Western-blotting"

Fig 3

Osteoclast differentiation detected by TRAP and bovine bone resorption lacuna"

Fig 4

c-fos mRNA, c-jun mRNA and CREB mRNA detected by real-time PCR"

Fig 5

Expression of c-Fos, c-Jun and CREB detected by Western-blotting"

[1] Pereira M, Petretto E, Gordon S , et alCommon signalling pathways in macrophage and osteoclast multinucleation[J]. J Cell Sci, 2018,131:11. pii:jcs216267.
[2] Yuan FL, Wu QY, Miao ZN , et al. Osteoclast-derived extracellular vesicles: novel regulators of osteoclastogenesis and osteoclast-osteoblasts communication in bone remodeling[J]. Front Physiol, 2018,9:628.
[3] Yao CH, Zhang P, Zhang L . Differential protein and mRNA expression of CaMKs during osteoclastogenesis and its functional implications[J]. Biochem Cell Biol, 2012,90(4):532-539.
[4] Kim JY, Cheon YH, Yoon KH , et al. Parthenolide inhibits osteoclast differentiation and bone resorbing activity by down-regulation of NFATc1 induction and c-Fos stability, during RANKL-mediated osteoclastogenesis[J]. BMB Rep, 2014,47(8):451-456.
[5] Mohamed SG, Sugiyama E, Shinoda K , et al. Interleukin-10 inhibits RANKL-mediated expression of NFATc1 in part via suppression of c-Fos and c-Jun in RAW264.7 cells and mouse bone marrow cells[J]. Bone, 2007,41(4):592-602.
doi: 10.1016/j.bone.2007.05.016
[6] Park KH, Gu DR, Jin SH , et al. Pueraria lobate inhibits RANKL-mediated osteoclastogenesis via dow-nregulation of CREB/PGC1β/c-Fos/NFATc1 signaling[J]. Am J Chin Med, 2017,45(8):1725-1744.
[7] 刘娟娟, 董伟, 戚孟春 , 等. 钙离子/钙调蛋白依赖性蛋白激酶Ⅱδ RNA干扰对下游基因表达及破骨细胞分化的影响[J]. 第三军医大学学报, 2017,39(1):22-27.
Liu JJ, Dong W, Qi MC , et al. Influence of CaMK Ⅱ δ RNA interference on expression of its downstream molecules and osteoclast differentiation[J]. J Third Mil Med Univ, 2017,39(1):22-27.
[8] Teitelbaum SL . Bone resorption by osteoclasts[J]. Science, 2000,289(5484):1504-1508.
[9] Matsuo K, Galson DL, Zhao C , et al. Nuclear factor of activated T-cells (NFAT) rescues osteoclastogenesis in precursors lacking c-Fos[J]. J Biol Chem, 2004,279(25):26475-26480.
[10] Matsuo K, Ray N . Osteoclasts, mononuclear phagocytes, and c-Fos: new insight into osteoimmunology[J]. Keio J Med, 2004,53(2):78-84.
[11] Nie S, Xu J, Zhang C , et al. Salicortin inhibits osteoclast differentiation and bone resorption by down-regulating JNK and NF-κB/NFATc1 signaling pathways[J]. Biochem Biophys Res Commun, 2016,470(1):61-67.
[12] 王想福, 孙凤歧, 石瑞芳 , 等. 靶向抑制PI3K对人体外周血破骨细胞分化p38/c-Fos信号通路调控的研究[J]. 中国骨质疏松杂志, 2016,22(1):49-52.
Wang XF, Sun FQ, Shi RF , et al. The regulation of p38/c-Fos signal pathway by targeting inhibition of PI3K in human peripheral osteoclasts[J]. Chin J Osteopor, 2016,22(1):49-52.
[13] Izawa T, Arakaki R, Mori H , et al. The nuclear receptor AhR controls bone homeostasis by regulating osteoclast differentiation via the RANK/c-Fos signaling Axis[J]. J Immunol, 2016,197(12):4639-4650.
[14] 王红美, 董伟, 戚孟春 , 等. CREB和ERK1/2在CaMKⅡδ调控破骨细胞分化中的作用[J]. 解放军医学杂志, 2018,43(2):91-95.
Wang HM, Dong W, Qi MC , et al. Regulation of osteoclast differentiation by CaMKⅡδ mediated through CREB and ERK1/2 signal pathways[J]. Med J Chin PLA, 2018,43(2):91-95.
[15] 陆大壮, 刘娟娟, 戚孟春 , 等. CaMKⅡδ在破骨细胞分化不同阶段表达规律的研究[J]. 中国病理生理杂志, 2016,32(10):1870-1874.
Lu DZ, Liu JJ, Qi MC , et al. Expression profiles of CaMKⅡδ at different stages of osteoclast differen-tiation[J]. Chin J Pathophysiol, 2016,32(10):1870-1874.
[1] Yu Yuelin,Kong Weidong. Research progress on the association between primary failure of tooth eruption and parathyroid hormone receptor 1 gene [J]. Int J Stomatol, 2023, 50(5): 573-580.
[2] Liu Tiqian,Liang Xing,Liu Weiqing,Li Xiaohong,Zhu Rui.. Research progress on the role and mechanism of occlusal trauma in the development of periodontitis [J]. Int J Stomatol, 2023, 50(1): 19-24.
[3] An Ning,Li Jiao,Mei Zhidan. Research progress on the osteoprotegerin/receptor activator of nuclear factor-κB/receptor activator of nuclear factor-κB ligand signaling pathway of tooth eruption [J]. Int J Stomatol, 2022, 49(1): 116-120.
[4] Lü Hui,Wang Hua,Sun Wen. T helper cell 17 and periodontitis related osteoimmunology [J]. Int J Stomatol, 2020, 47(6): 661-668.
[5] Fu Shijin,Zeng Kan,Li Xin,Yang Jing,Wang Chenglin,Ye Ling. Preliminary study on osteoprotegerin/receptor activator of nuclear factor-κB ligand expression in mandible and femur on site selectivity of bone metastasis of lung cancer cells [J]. Int J Stomatol, 2020, 47(5): 538-546.
[6] Sun Jianwei,Lei Lihong,Tan Jingyi,Chen Lili. Regulation of osteoimmunology by MicroRNA 155 and research progress of its possible mechanism in periodontitis [J]. Int J Stomatol, 2020, 47(5): 607-615.
[7] Yang Peipei,Yang Yuchen,Zhang Qiang. Advances in the mechanism and effect of nicotine on alveolar osteoclasts [J]. Int J Stomatol, 2020, 47(5): 616-620.
[8] Zhu Junjin,Zhou Jiaqi,Wu Yingying. Function of autophagy induced by mammalian target of rapamycin complex 1 in bone metabolism [J]. Int J Stomatol, 2020, 47(1): 84-89.
[9] Lu Kexin,Zhang Diya,Wu Yanmin. Research progress of protease-activated receptors on different types of cells in periodontal tissue [J]. Int J Stomatol, 2019, 46(6): 657-662.
[10] Wei Hu,Yifan Wang,Yifang Yuan,Ying Li,Bin Guo. Research progress on regulatory mechanism of the circadian clock genes on osteogenesis and bone resorption [J]. Int J Stomatol, 2019, 46(3): 302-307.
[11] Xin Gao,Rongsheng Zeng. Research progress on osteoprotegerin in oral science [J]. Int J Stomatol, 2019, 46(3): 316-319.
[12] Cui Yue, Jiang Huan, Hu Min.. The relationship between osteoclastic protein-tyrosine phosphatase and orthodontic treatment induced root resorption [J]. Inter J Stomatol, 2017, 44(1): 87-91.
[13] Hou Yubo1, Liu Xinchan2, Yu Haiyan1, Cui Leihua3, Yu Weixian4. Effect of gingipains on osteoclasts and osteoblasts [J]. Inter J Stomatol, 2016, 43(5): 609-613.
[14] Tan Xiujun, Ye Ling, Wang Chenglin. Mechanisms of c-Jun N-terminal kinase signal transduction pathway regulating cell migration [J]. Inter J Stomatol, 2015, 42(5): 586-591.
[15] Zhou Zheng, Zhao Changming, Jiao Kai, Wang Meiqing. Regulatory role of sympathetic nervous system–adrenergic receptor on bone remodeling [J]. Inter J Stomatol, 2015, 42(3): 348-351.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[2] . [J]. Foreign Med Sci: Stomatol, 1999, 26(05): .
[3] . [J]. Foreign Med Sci: Stomatol, 1999, 26(05): .
[4] . [J]. Foreign Med Sci: Stomatol, 1999, 26(05): .
[5] . [J]. Foreign Med Sci: Stomatol, 1999, 26(05): .
[6] . [J]. Foreign Med Sci: Stomatol, 1999, 26(04): .
[7] . [J]. Foreign Med Sci: Stomatol, 2005, 32(06): 458 -460 .
[8] . [J]. Foreign Med Sci: Stomatol, 2005, 32(06): 452 -454 .
[9] . [J]. Inter J Stomatol, 2008, 35(S1): .
[10] . [J]. Inter J Stomatol, 2008, 35(S1): .