Osteoporosis International

, Volume 22, Issue 7, pp 2175–2186 | Cite as

Quantitative proteomic analysis of dexamethasone-induced effects on osteoblast differentiation, proliferation, and apoptosis in MC3T3-E1 cells using SILAC

Original Article



The impairment of osteoblast differentiation is one cause of the glucocorticoid-induced osteoporosis (GCOP). The quantitative proteomic analysis of the dexamethasone (DEX)-induced effects of osteoblast differentiation, proliferation, and apoptosis using stable-isotope labeling by amino acids in cell culture (SILAC) demonstrated drastic changes of some key proteins in MC3T3-E1 cells.


The impairment of osteoblast differentiation is one of the main explanations of GCOP. SILAC enables accurate quantitative proteomic analysis of protein changes in cells to explore the underlying mechanism of GCOP.


Osteoprogenitor MC3T3-E1 cells were treated with or without 10−6 M DEX for 7 days, and the differentiation ability, proliferation, and apoptosis of the cells were measured. The protein level changes were analyzed using SILAC and liquid chromatography-coupled tandem mass spectrometry.


In this study, 10−6 M DEX inhibited both osteoblast differentiation and proliferation but induced apoptosis in osteoprogenitor MC3T3-E1 cells on day 7. We found that 10−6 M DEX increased the levels of tubulins (TUBA1A, TUBB2B, and TUBB5), IQGAP1, S100 proteins (S100A11, S100A6, S100A4, and S100A10), myosin proteins (MYH9 and MYH11), and apoptosis and stress proteins, while inhibited the protein levels of ATP synthases (ATP5O, ATP5H, ATP5A1, and ATP5F1), G3BP-1, and Ras-related proteins (Rab-1A, Rab-2A, and Rab-7) in MC3T3-E1 cells.


Several members of the ATP synthases, myosin proteins, small GTPase superfamily, and S100 proteins may participate in functional inhibition of osteoblast progenitor cells by GCs. Such protein expression changes may be of pathological significance in coping with GCOP.


Apoptosis Dexamethasone Osteoblast differentiation Proliferation Proteomics SILAC 



Actin, alpha skeletal muscle


Actin, cytoplasmic 1


Alkaline phosphatase


Annexin A1


Annexin A8


ATP synthase subunit alpha, mitochondrial precursor


ATP synthase B chain, mitochondrial precursor


ATP synthase D chain, mitochondrial


ATP synthase O subunit, mitochondrial precursor


Apoptosis regulator BAX




Ras GTPase-activating protein-binding protein 1


Glyceraldehyde-3-phosphate dehydrogenase


GC-induced osteoporosis




Heat shock 70 kDa protein 4


Heat shock protein HSP 90-alpha


Ras GTPase-activating-like protein 1


Integrator complex subunit 3


Ras-related protein


Stable-isotope labeling by amino acids in cell culture


Tubulin alpha-1 chain


Tubulin beta-2B chain


Tubulin beta-5 chain


Liquid chromatography-coupled tandem mass spectrometry


3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


Myosin 9


Myb-binding protein 1A


Isoform 1 of myosin 11


Programmed cell death 6-interacting protein


Programmed cell death 6


Protein Interrogation of Gene Ontology and KEGG databases


Polymerase I and transcript release factor


Peptidylprolyl isomerase B


Transitional endoplasmic reticulum ATPase



This work is supported by the NIH (Grant DE 018385), the National Natural Science Foundation of China (30950019), and the Medicine and Health Research Fund of Zhejiang Province, China (2009B166). We thank Chantal M. Sottas for the technical support.

Conflicts of interest



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Copyright information

© International Osteoporosis Foundation and National Osteoporosis Foundation 2010

Authors and Affiliations

  1. 1.Population CouncilNew YorkUSA
  2. 2.Orthopedic DepartmentTaizhou Hospital, Wenzhou Medical CollegeLinhaiChina
  3. 3.Proteomics Resource CenterThe Rockefeller UniversityNew YorkUSA
  4. 4.The Second Affiliated HospitalWenzhou Medical CollegeWenzhouChina

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