title: Expression and regulation of amphiregulin in Gsα-mutated human bone marrow stromal cells of fibrous dysplasia of mandible.
abstract: OBJECTIVE
  Fibrous dysplasia (FD) is a focal bone lesion composed primarily of immature bone marrow stromal cells along with spicules of immature woven bone. However, cellular differentiation and proliferation in mutant human bone marrow stromal cells (hBMSCs) of FD have not been fully elucidated. Therefore, the aim of this study was to investigate the occurrence of G(s)α mutation at the Arg(201) codon in hBMSCs and human trabecular bone cells (hTBCs, osteoblast-like cells). In addition, we evaluated the gene expression and protein secretion of amphiregulin from hBMSCs and hTBCs and assessed the biologic activity and possible mechanism of amphiregulin in our system.
  
  
  METHODS
  Mutant hBMSCs from FD patients and hTBCs from disease-free bone specimens of the same patient were successfully cultured. We studied the G(s)α mutations at the Arg(201) codon by means of polymerase chain reaction (PCR)-restriction fragment length polymorphism. Gene expression and protein secretion of amphiregulin in hBMSCs and hTBCs was confirmed by reverse-transcription (RT) PCR and Western blotting analysis. The modulation proliferation and possible mechanism of the exogenous addition of amphiregulin and epidermal growth factor receptor tyrosine kinase inhibitor (AG-1478) were assessed by using Wst-1 assays.
  
  
  RESULTS
  The G(s)α mutations in clonal adherent mutant hBMSCs and hTBCs were successfully identified. We identified amphiregulin to be highly expressed in hBMSCs compared with hTBCs. The growth of hBMSCs was stimulated by the exogenous addition of amphiregulin and inhibited by AG-1478.
  
  
  CONCLUSIONS
  The G(s)α-mutant hBMSCs were successfully identified, and amphiregulin may play a significant role in the proliferation of hBMSCs.
authors: Department of Orthodontics, Provincial Hospital, Affiliated to Shandong University, Ji'nan, China.
authors: Shanshan
authors: S
authors: Jia
authors: Jianbin
authors: J
authors: Yu
authors: Dongsheng
authors: D
authors: Zhang
authors: Peihui
authors: P
authors: Zheng
authors: Shizhou
authors: S
authors: Zhang
authors: Li
authors: L
authors: Ma
authors: Guijun
authors: G
authors: Liu
authors: Shengfeng
authors: S
authors: Li
mesh_terms: D000293:Adolescent; D019943:Amino Acid Substitution; D066258:Amphiregulin; D001120:Arginine / Q000235:genetics; D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004396:Coloring Agents; D004252:DNA Mutational Analysis; D066255:EGF Family of Proteins; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005260:Female; D005357:Fibrous Dysplasia of Bone / Q000235:genetics* / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics*; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D008336:Mandibular Diseases / Q000235:genetics*; D020125:Mutation, Missense; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D012150:Polymorphism, Restriction Fragment Length; D011799:Quinazolines; D015398:Signal Transduction; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism; D013778:Tetrazolium Salts; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; C587848:AREG protein, human; D066258:Amphiregulin; D004396:Coloring Agents; D066255:EGF Family of Proteins; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D011799:Quinazolines; D013778:Tetrazolium Salts; D020032:Tyrphostins; C101044:RTKI cpd; D001120:Arginine; D066246:ErbB Receptors; D019205:GTP-Binding Protein alpha Subunits, Gs