title: Impact of oxidative stress on the cytoskeleton of pancreatic epithelial cells.
abstract: In the present study the effect of reactive oxygen species on the morphological changes of pancreatic epithelial cells in a three-dimensional culture system was investigated. In addition, the expression of signaling molecules during this process was determined. Matrigel™ was used to construct a three-dimensional culture model of pancreatic epithelial and cancer cells. The cultured cells were stimulated with 1 or 200 μmol/l H2O2(a typical reactive oxygen species), and the morphological changes were then evaluated after 15 min, 1 h and 4 h. The cytoskeleton of the cells was observed using laser scanning confocal microscopy with immunofluorescence staining. In addition, the nuclear content of nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) was detected using ELISA. The results demonstrated that treatment with 200 μmol/l H2O2induced cell contraction after 15 min, and cell morphology recovered after 1 h; however, cell size was reduced after 4 h. Consequently, intracellular actin and microtubules were rapidly lost following H2O2treatment, and the cytoskeleton became indistinct and eventually disintegrated after 4 h. Similar observations were noted for the normal pancreatic epithelial and cancer cells. By contrast, treatment with 1 μmol/l H2O2did not affect the morphology and cytoskeleton of pancreatic epithelial cells. In addition, 200 μmol/l H2O2treatment increased the activity of NF-κB gradually, while 1 μmol/l H2O2treatment was found to have little impact on the activity of NF-κB. Therefore, it was demonstrated that oxidative stress can induce the early onset of reversible cell contraction and cytoskeleton depolarization in pancreatic epithelial cells, and can increase NF-κB expression.
authors.affiliation: Department of General Surgery, Wuhan General Hospital of Guangzhou Military Command, Wuhan, Hubei 430070, P.R. China.
authors.forename: Wei-Guo
authors.initials: WG
authors.lastname: Hu
authors.affiliation: Department of General Surgery, Wuhan General Hospital of Guangzhou Military Command, Wuhan, Hubei 430070, P.R. China.
authors.forename: Qi-Ping
authors.initials: QP
authors.lastname: Lu
keywords: nuclear factor κ-light-chain-enhancer of activated B cells; oxidative stress; pancreatic disease