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Spatial--temporal distribution of glycogen synthase kinase 3#beta# and adenomatous polyposis coli protein are involved in the injury and repair of airway epithelial cells induced by scratching

Zhu Min, Li Jiansha, Tian Dan, Ma Yan, Li Naping, Wu Renliang

Department of Pathology,Tongji Medical College,Huazhong University of Science and Technology,and Key Laboratory of Pulmonary Disease of Ministry of Health of China.Wuhan 430030,Hubei

Abstract

To investigate the roles of glycogen synthase kinase 3#beta#(GSK3#beta#) and adenomatous polyposis coli (APC) protein in wound repair of airway epithelial cells (AECs), we established a wound model of airway epithelium in vitro. Then the following tests were undertaken: (1) Western blot was used to detect the levels of total GSK3#beta# and phosphorylated GSK3#beta# in human bronchial epithelial (16HBE) cells; (2) The localizations of APC protein was observed by using immunofluorescence technique; (3) Immunoprecipitation was used to investigate the relationship between APC protein and GSK3#beta# during the repair of 16HBE cells. The results showed: (1) The level of phosphorylated GSK3#beta# increased 0.5 h after scratching (P<0.05), reached a maximum at 6 h (P<0.05), and maintained until 12 h, while the total levels of GSK3#beta# remained constant; (2) Results of immunofluorescence study showed that the APC protein clustered with tubulin in the region of the migrating leading cells 6 h after scratching, which was dissimilar with the localization of the protein in the control group; (3) GSK3#beta# and APC protein were immunoprecipitated and analysed on SDS-PAGE. We found that GSK3#beta# and APC protein were precipitated, indicating that the two proteins existed in a complex. After scratching, significant dissociation occurred between the proteins. Taken together, we conclude that scratching caused a decrease in phosphorylation of GSK3#beta#, and that reduced phosphorylation of GSK3#beta# promoted the APC protein to bind to the plus ends of microtubules and stabilize the growing ends. These observations suggest that APC protein and GSK3#beta# may synergistically play an important role in the repair of airway epithelium.

Key words: glycogen synthase kinase 3#beta#;adenomatous polyposis coli protein;airway epithelial cells;injury and repair

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Citing This Article：

Zhu Min, Li Jiansha, Tian Dan, Ma Yan, Li Naping, Wu Renliang. Spatial--temporal distribution of glycogen synthase kinase 3#beta# and adenomatous polyposis coli protein are involved in the injury and repair of airway epithelial cells induced by scratching. Acta Physiol Sin 2007; 59 (2): 197-203 (in Chinese with English abstract).