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2015 ; 16
(1
): 72
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Fibroblast-epithelial cell interactions drive epithelial-mesenchymal transition
differently in cells from normal and COPD patients
#MMPMID26081431
Nishioka M
; Venkatesan N
; Dessalle K
; Mogas A
; Kyoh S
; Lin TY
; Nair P
; Baglole CJ
; Eidelman DH
; Ludwig MS
; Hamid Q
Respir Res
2015[Jun]; 16
(1
): 72
PMID26081431
show ga
BACKGROUND: Epithelial-to-mesenchymal transition (EMT), which involves changes in
cellular morphology of highly polarized epithelial cells and the gain of
mesenchymal cell phenotype with migratory and invasive capacities, is implicated
in smoking-related chronic obstructive pulmonary disease (COPD). However, the
interactions of fibroblasts and epithelial cells and the participation of
fibroblasts in the EMT processes in COPD are poorly understood. Here, we
investigated the hypothesis that EMT is active in human bronchial epithelial
(HBE) cells of COPD patients, and that mediators secreted by lung fibroblasts
from COPD patients induce EMT. METHODS: Primary HBE cells from normal subjects
and COPD patients were purchased from LONZA. HLFs were derived from resected lung
obtained from normal (N) and COPD (D) subjects and their conditioned medium (CM)
was collected after 2-day culture in serum-free medium. The expression of
epithelial and mesenchymal markers as well as EMT-related transcription factors
in lung biopsies, and in HBE cells following stimulation with CM from both normal
human lung fibroblasts (NHLF) and COPD human lung fibroblasts (DHLF) was
evaluated by immunohistochemistry, qRT-PCR and western blot. RESULTS: Basal mRNA
expression of mesenchymal markers and EMT-related transcription factors were
increased in DHBE cells compared to normal human bronchial epithelial cells
(NHBE) cells as well as in COPD lungs. CM from NHLF significantly induced
vimentin expression in both NHBE and COPD human bronchial epithelial cells (DHBE)
cells, but only increased N-cadherin expression in DHBE cells. CM from NHLF
significantly induced Twist1 and Twist2 expression in NHBE cells and increased
Snai2 (Slug) expression in DHBE cells. While CM from NHLF had no effect on such
EMT markers, CM from DHLF significantly increased the protein expression of
E-cadherin and vimentin in NHBE cells compared to control. N-cadherin expression
was upregulated to a greater degree in NHBE cells than DHBE cells. Only CM from
DHLF significantly increased E-/N-cadherin ratio in DHBE cells. CONCLUSIONS: Our
results suggest that DHBE cells have partially undergone EMT under baseline
conditions. DHLF-CM promoted EMT in NHBE, suggesting that interactions between
fibroblast and epithelial cells may play an important role in the EMT process in
COPD.