title: [Phosphorylation of ezrin Tyr477 is critical in invasion of breast cancer cells stimulated by precursor of nerve growth factor].
abstract: OBJECTIVE
  To investigate the effect of precursor of nerve growth factor (proNGF) in promoting invasion of breast cancer cells and its relation with ezrin expression and phosphorylation of ezrin Thr567 and Tyr477.
  
  
  METHODS
  Human breast cancer cell lines MDA-MB-231 and MCF-7 were stimulated by gradient concentrations of proNGF (0, 2.5, 5 and 10 ng/mL) for 16 h, and the invasion of the cells was assessed with Transwell assay. The expression of ezrin and the phosphorylation of ezrin Thr567 and ezrin Tyr477 in the treated cells were examined by Western blotting. MDA-MB-231 cells were transfected with pEnter-His-ezrinY477F (a dominant negative mutant) to study the role of phosphrylation of ezrin Tyr477 in the invasion of breast cancer cell stimulated by proNGF.
  
  
  RESULTS
  proNGF significantly promoted MDA-MB-231 and MCF-7 cell invasion in a concentration-dependent manner (P<0.05), and concentration- and time-dependently increased the phosphorylation of ezrin Tyr477 (P<0.05) without affecting the expression of ezrin or the phosphorylation of ezrin Thr567. The specific inhibitor of src, SKI-606, significantly inhibited the phosphorylation of ezrin Tyr477 induced by proNGF. Transfection with pEnter-His- ezrinY477F inhibited proNGF-induced invasion and phosphorylation of ezrin Tyr477 in MDA-MB-231 cells (P<0.05).
  
  
  CONCLUSIONS
  Phosphorylation of ezrin Tyr477 plays a critical role in the invasion of breast cancer cells stimulated by proNGF via proNGF/src/ezrin Tyr477 pathway.
authors.affiliation: Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China.E-mail: 514380534@qq.com.
authors.forename: Fu-Xi
authors.initials: FX
authors.lastname: Li
authors.forename: Wen-Feng
authors.initials: WF
authors.lastname: Shao
authors.forename: Rui
authors.initials: R
authors.lastname: Tang
authors.forename: Xiao-Ran
authors.initials: XR
authors.lastname: Yu
authors.forename: Qiao-Sheng
authors.initials: QS
authors.lastname: Wen
authors.forename: Yan-Lin
authors.initials: YL
authors.lastname: Yu
authors.forename: Jing-Bo
authors.initials: JB
authors.lastname: Xiong
mesh_terms: D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000737:chemistry*; D006801:Humans; D061986:MCF-7 Cells; D009361:Neoplasm Invasiveness; D020932:Nerve Growth Factor / Q000494:pharmacology*; D010766:Phosphorylation; D015398:Signal Transduction; D014162:Transfection; D014443:Tyrosine; D000099149:Ezrin
chemical_list: D003598:Cytoskeletal Proteins; D020932:Nerve Growth Factor; D014443:Tyrosine; D000099149:Ezrin