Differentiation of gastric surface mucous cells (GSM06) induced by air–liquid interface is regulated partly through mitogen-activated protein kinase pathway

Background and Aim: The aim of the present study was to examine the role of mitogen-activated protein (MAP) kinase pathway on gastric surface epithelium using an established cell culture model in which differentiation is promoted in GSM06 cells by air–liquid interface.

Methods: A double-dish culture system of mouse gastric surface mucous cell line GSM06 in Ham's F12 medium supplemented with 10% fetal calf serum and 50 μg/mL gentamicin at 37°C in a humidified atmosphere of 5% CO₂ in air was used for an air–liquid interface. Culture cells were examined on histology, cell proliferation was evaluated by bromodeoxy-uridine (BrdU) uptake, and western blot analysis of extracellular signal-regulated kinase (ERK)1/2 and phosphate ERK1/2. On day 3, U0126, an inhibitor of MAP kinase kinase (MEK), was added to medium of incubated cells.

Results: GSM06 cells were differentiated with an air–liquid interface for 3 weeks. Compared to immersion control culture, phosphorylated ERK 1/2 expression increased significantly. This increase was completely suppressed with U0126, and tall columnar cells developed by air–liquid interface in GSM06 were not observed in U0126-treated cells. Increase in BrdU uptake with air–liquid interface was suppressed by U0126.

Conclusion: These results suggested that MAP kinase signaling, activated by air–liquid interface, was, at least in part, related to cell differentiation in GSM06 cells induced by air–liquid interface.