A proteomic approach to mimic fibrosis disease evolvement by an in vitro cell line

Subepithelial fibrosis in asthma involves an increase in the thickening of the lamina reticularis and is due to increased deposition of collagen I, II I and V, and fibronectin. The cause of the thickening of the reticular laye r is not known in detail, however, it is proposed to be caused by bronchial myofibroblasts. The transformation of fibroblasts to myofibroblasts may be contributed by inflammatory cytokines. In this paper we have studied and c ompared in vivo tissue material with a human fibroblast target cell. A norm al primary fetal fibroblast cell line and HFL-1 (human fibroblast lurg cell s) were used as a comparison between fibroblasts from human central biopsie s regarding morphology and cell proliferation. Both cell morphology and cel l proliferation rate was similar between the different set of cell cultures . Furthermore, it could be concluded that fibroblasts cultures from patient s with asthma were surrounded by more extracellular matrix molecules compar ed to the primary cell line HFL-1, which may mimic the in vivo situation du ring formation of fibrosis. We wanted to investigate if differential protei n display by two-dimensional (2-D) gel electrophoresis and subsequent prote in identification by matrix assisted laser desorption/ionization-time of fl ight (MALDI-TOF)-mass spectrometry could reveal proteins induced by cytokin e stimulation that can be correlated to the transformation of normal human fetal lungs cells into a more myofibroblast like phenotype. After stimulati on with transforming growth factor-beta (TGF-beta) several myofibroblast ma rkers were found to be regulated. Especially cytoskeletal and cytoskeletal- associated proteins like actin isoforms and tropomyosin, proteins that are responsible for contraction as well as transportation of extra cellular mat rix proteins, which are overproduced in the formation of fibrosis. These re sults indicate that TGF-beta, which is increased in a fibrotic process, par ticipates in the transformation of fibroblasts to myofibroblasts.