表皮生长因子对结肠细胞中紧密连接蛋白表达和磷酸化的影响

Effect of epidermal growth factor on the expression and phosphorylation of tight junction protein in colonic cells

目的探讨表皮生长因子(EGF)对T84人结肠癌细胞中紧密连接(TJs)蛋白表达和磷酸化的影响。方法培养T84细胞至单层融合状态,对照组和实验组分别加入二甲基亚砜(DMSO)和EGF,观察TJs蛋白claudin亚型、occludin和Zo-1的表达差异和磷酸化,以及损伤/修复的效果。结果与对照组比较,EGF干预组细胞在48 h后磷酸化claudin-3水平下降,磷酸化claudin-5、claudin-7水平升高,非磷酸化claudin-1、claudin-3水平升高(P <0. 05),非磷酸化claudin-7水平没有变化;非磷酸化claudin-5水平在72 h后下降(P <0. 05)。非磷酸化Zo-1水平在EGF干预24 h后即升高,72 h后有所下降(P <0. 05)。划痕后24 h及48 h,EGF干预组细胞迁移率明显高于对照组(P <0. 05)。结论 EGF可以影响肠上皮细胞间TJs蛋白的表达和磷酸化状态,并促进损伤肠上皮细胞的修复。

Objective To investigate the effect of epidermal growth factor(EGF)on the tight junction protein expression and phosphorylation in T84 colonic cell.Methods T84 cells were cultured to monolayer state,and DMSO and EGF were added to the control and experimental groups respectively.The expression and phosphorylation of tight junction proteins,claudins,occludin and Zo-1,and the damage/repair rate in T84 cells were observed after treatment.Results Compared with the control group,EGF decreased the phosphorylation of claudin-3,increased the phosphorylation of claudin-5 and claudin-7,increased the levels of non-phosphorylated claudin-1,claudin-3 at 48 hours after intervention as well(P<0.05).The level of non-phosphorylated claudin-7 did not change at 48 hours after EGF intervention,but the level of non-phosphorylated claudin-5 was decreased at 72 hours after EGF intervention(P<0.05).The level of non-phosphorylated Zo-1 was increased 24 hours after EGF intervention,but decreased at 72 hours(P<0.05).Compared with the control group,the migration of EGF group was significantly higher than that of control group,24 hours and 48 hours after scratches(P<0.05).Conclusion EGF can affect the expression and phosphorylation of tight junction proteins,and promote the damage/repair of intestinal epithelial cells.