title: Independent of ErbB1 gene copy number, EGF stimulates migration but is not associated with cell proliferation in non-small cell lung cancer.
abstract: BACKGROUND
  Lung cancer often exhibits molecular changes, such as the overexpression of the ErbB1 gene. ErbB1 encodes epidermal growth factor receptor (EGFR), a tyrosine kinase receptor, involved mainly in cell proliferation and survival. EGFR overexpression has been associated with more aggressive disease, poor prognosis, low survival rate and low response to therapy. ErbB1 amplification and mutation are associated with tumor development and are implicated in ineffective treatment. The aim of the present study was to investigate whether the ErbB1 copy number affects EGFR expression, cell proliferation or cell migration by comparing two different cell lines.
  
  
  METHODS
  The copies of ErbB1 gene was evaluated by FISH. Immunofluorescence and Western blotting were performed to determine location and expression of proteins mentioned in the present study. Proliferation was studied by flow cytometry and cell migration by wound healing assay and time lapse.
  
  
  RESULTS
  We investigated the activation and function of EGFR in the A549 and HK2 lung cancer cell lines, which contain 3 and 6 copies of ErbB1, respectively. The expression of EGFR was lower in the HK2 cell line. EGFR was activated after stimulation with EGF in both cell lines, but this activation did not promote differences in cellular proliferation when compared to control cells. Inhibiting EGFR with AG1478 did not modify cellular proliferation, confirming previous data. However, we observed morphological alterations, changes in microfilament organization and increased cell migration upon EGF stimulation. However, these effects did not seem to be consequence of an epithelial-mesenchymal transition.
  
  
  CONCLUSIONS
  EGFR expression did not appear to be associated to the ErbB1 gene copy number, and neither of these aspects appeared to affect cell proliferation. However, EGFR activation by EGF resulted in cell migration stimulation in both cell lines.
authors: Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of Sao Paulo, Av, Prof, Lineu Prestes, 1524, Butantã, São Paulo, SP 05508-000, Brazil. gmmsante@usp.br.
authors: Camila
authors: C
authors: Lauand
authors: Paula
authors: P
authors: Rezende-Teixeira
authors: Beatriz Araújo
authors: BA
authors: Cortez
authors: Evandro Luís de Oliveira
authors: EL
authors: Niero
authors: Gláucia Maria
authors: GM
authors: Machado-Santelli
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