【摘要】 为探讨糖原合成酶激酶3(glycogen synthase kinase 3,GSK3)在气道(气管和支气管)上皮细胞鳞状分化中的作用,培养原代猪气道上皮细胞,用GSK3的高度选择性抑制剂氯化锂处理,观察细胞形态变化,用Western blot检测β-连环素、磷酸化GSK3和鳞状分化标记物外皮蛋白的表达、RT-PCR检测鳞状分化标记物小脯氨酸丰富蛋白mRNA的表达、荧光素酶报告基因分析β-连环素/Tcf信号的激活状态。结果显示,锂能诱导猪气道上皮细胞出现鳞状形态、增加小脯氨酸丰富蛋白mRNA和外皮蛋白的表达、促进GSK3的抑制性丝氨酸磷酸化和β-连环素的细胞核内转位;锂能激活β-连环素/Tcf信号,但该作用出现于鳞状分化标记物增加之后。上述结果提示,GSK3可能参与猪气道上皮细胞的鳞状分化。更多还原

【Abstract】 To investigate if glycogen synthase kinase 3 (GSK3) is involved in squamous differentiation of airway (tracheobronchial)epithelial cells, primary pig airway epithelial cells were treated with lithium chloride, a highly selective inhibitor of GSK3. Change inmorphology of cells was monitored under microscopy, and expression of β-catenin, phosphorylated GSK3 and involucrin, a squamousdifferentiation marker, were dectected by Western blotting, while expression of mRNA of another squamous differentiation marker,small proline-rich protein, was detected by RT-PCR. Further, luciferase reporter assay was used to assess the activation of β-catenin/Tcf signaling. The results demonstrated that lithium was able to induce a squamous morphology of the cells, and to enhance theexpression of involucrin and small proline-rich protein mRNA. Moreover, lithium increased inhibitory phosphorylation of GSK3,augmented nuclear translocation of β-catenin in a dose- and time-dependent manner. Activation of β-catenin/Tcf signaling was observedafter the elevation of squamous differentiation markers. Taken together, these data suggest that GSK3 is possibly involved in squamousdifferentiation of pig airway epithelial cells.更多还原