Abstract:Objective The abnormal proliferation of osteoblasts leads to decreased bone mass,which is an important reason for the occurrence and development of osteoporosis.A large number of studies have found that Orai1 plays a very important role in regulating osteoblast proliferation and apoptosis.STIM1 is an important protein that can activate Orai1.However,there is no relevant report on whether STIM1 is involved in the regulation of osteoblast proliferation The purpose of this study was to determine the role of STIM1 in regulating osteoblast proliferation in osteoblasts.Methods After STIM1 knockdown,cell proliferation and cell cycle,the transcription of CyclinD1,CyclinE,CDK4 and CDK6,and Ras signaling pathway activity were tested to identify whether STIM1 is involved in the regulation of osteoblast proliferations.Results STIM1 was knockdown successfully after STIM1 siRNA transfected in MC3T3-E1 cells(P<0.05).We found that the proliferation were decreased and cell cycle were arrested after STIM1 knockdown(P<0.05).Furthermore,we found that the transcription of CyclinD1 was decreased after STIM1 knockdown(P<0.05).We also found that Ras signaling pathway activity were inhibited(P<0.05).The inhibition of Ras signaling pathway activity by ars-853 significantly inhibited the role of STIM1 in promoting the proliferation of osteoblasts(P<0.05).Conclusion The results identified that STIM1 may activate Ras signaling pathway,thus regulating osteoblast proliferation.
[1]Phetfong J,Sanvoranart T,Nartprayut K,et al.Osteoporosis:the current status of mesenchymal stem cell-based therapy[J].Cell Mol Biol Lett,2016,21(8):12.
[2]Hu LF,Yin C,Zhao F,et al.Mesenchymal stem cells:cell fate decision to osteoblast or adipocyte and application in osteoporosis treatment[J].Int J Mol Sci,2018,19(2):360.
[3]Jiang ZW,Li Z,Zhang W,et al.Dietary natural N-acetyl-d-glucosamine prevents bone loss in ovariectomized rat model of postmenopausal osteoporosis[J].Molecules,2018,23(9):2302.
[4]Tarantino U,Iolascon G,Cianferotti L,et al.Clinical guidelines for the prevention and treatment of osteoporosis:summary statements and recommendations from the Italian Society for Orthopaedics and Traumatology[J].J Orthop Traumatol,2017,18(Suppl 1):3-36.
[5]Zhao B,Zhao WQ,Wang YQ,et al.Prior administration of vitamin K2 improves the therapeutic effects of zoledronic acid in ovariectomized rats by antagonizing zoledronic acidinduced inhibition of osteoblasts proliferation and mineralization[J].PLoS One,2018,13(8):e0202269.
[6]Chakraborty S,Bipan KD,Chorna T,et al.Mutant IP3 receptors attenuate store-operated Ca2+ entry by destabilizing STIM-Orai interactions in Drosophila neurons[J].J Cell Sci,2016,129(20):3903-3910.
[7]Choi H,Srikanth S,Atti E,et al.Deletion of Orai1 leads to bone loss aggravated with aging and impairs function of osteoblast lineage cells[J].Bone Rep,2018,8(6):147-155.
[8]Cai XG,Robert M,Nwokonko NA.Pore properties of Orai1 calcium channel dimers and their activation by the STIM1 ER calcium sensor[J].J Biol Chem,2018,293(33):12962-12974.
[9]Zheng HF,Bernard TD,Earley S,et al.SOCE mediated by STIM and Orai is essential for pacemaker activity in the interstitial cells of Cajal in the gastrointestinal tract[J].Sanders Sci Signal,2018,11(534):eaaq0918.
[10]Liu JR,Xu GM,Shi XS,et al.Low temperature plasma promoting fibroblast proliferation by activating the NF-κB pathway and increasing cyclinD1 expression[J].Sci Rep,2017,7(9):11698.
[11]Casimiro MC,Sante GD,Ju XM et al.Cyclin D1 promotes androgen-dependent DNA damage repair in prostate cancer cells[J].Cancer Res,2016,76(2):329-338.
[12]Colleoni B,Paternot S,Pita JM,et al.JNKs function as CDK4-activating kinases by phosphorylating CDK4 and p21[J].Oncogene,2017,36(30):4349-4361.
[13]Chen HB,Xu XW,Wang GP,et al.CDK4 protein is degraded by anaphase-promoting complex/cyclosome in mitosis and reaccumulates in early G1 phase to initiate a new cell cycle in HeLa cells[J].J Biol Chem,2017,292(24):10131-10141.
[14]Peng S,Zhou G,Luk KD,et al.Strontium promotes osteogenic differentiation of mesenchymal stem cells through the Ras/MAPK signaling pathway[J].Cell Physiol Biochem,2009,23(1-3):165-174.
[15]Keiper M,Stope MB,Szatkowski D,et al.Epac-and Ca2+-controlled activation of Ras and extracellular signal-regulated kinases by Gs-coupled receptors[J].J Biol Chem,2004,279(45):46497-46508.
[16]Wajdner HE,Farrington J,Barnard C,et al.Orai and TRPC channel characterization in FcεRI-mediated calcium signaling and mediator secretion in human mast cells[J].Physiol Rep,2017,5(5):e13166.
[17]Staehlke S,Rebl H,Finke B,et al.Enhanced calcium ion mobilization in osteoblasts on amino group containing plasma polymer nanolayer[J].Cell Biosci,2018,8(3):22.
[18]Wang H,Sun W,Ma JQ,et al.Polycystin-1 mediates mechanical strain-induced osteoblastic mechanoresponses via potentiation of intracellular calcium and Akt/β-Catenin pathway[J].PLoS One,2014,9(3):e91730.
[19]DelaineSmith RM,Sittichokechaiwut A,Reilly GC.Primary cilia respond to fluid shear stress and mediate flow-induced calcium deposition in osteoblasts[J].FASEB J,2014,28(1):430-439.
[20]Martineau X,Abed E,Martel-Pelletier J,et al.Alteration of Wnt5a expression and of the non-canonical Wnt/PCP and Wnt/PKC-Ca2+ pathways in human osteoarthritis osteoblasts[J].PLoS One,2017,12(8):e0180711.
[21]Cabral WA,Ishikawa M,Garten M,et al.Absence of the ER cation channel TMEM38B/TRIC-B disrupts intracellular calcium homeostasis and dysregulates collagen synthesis in recessive osteogenesis imperfecta[J].PLoS Genet,2016,12(7):e1006156.
[22]Peterlik M,Kállay E,Heide S.Calcium nutrition and extracellular calcium sensing:Relevance for the pathogenesis of osteoporosis,cancer and cardiovascular diseases[J].Cross Nutrients,2013,5(1):302-327.
[23]Min PL,Quarles D.Osteoblast calcium-sensing receptor has characteristics of ANF/7TM receptors[J].J Cell Biochem,2005,95(6):1081-1092.
[24]Tharmalingam S,David R.The Calcium-sensing receptor and integrins in cellular differentiation and migration[J].Hampson Front Physiol,2016,7(5):190.
[25]Tang J,Liao Y,He SY,et al.Autocrine parathyroid hormone-like hormone promotes intrahepatic cholangiocarcinoma cell proliferation via increased ERK/JNK-ATF2-cyclinD1 signaling[J].J Transl Med,2017,15(11):238.
[26]Yang ZF,Kim S,Mahajan S,et al.Phospholipase Cγ1 (PLCγ1) controls osteoclast numbers via colony-stimulating factor 1 (CSF-1)-dependent diacylglycerol/β-catenin/cyclinD1 pathway[J].J Biol Chem,2017,292(4):1178-1186.
[27]Riverso M,Montagnani V,Stecca B.KLF4 is regulated by RAS/RAF/MEK/ERK signaling through E2F1 and promotes melanoma cell growth[J].Oncogene,2017,36(23):3322-3333.