陈文书, 吴人亮, 田丹, 王曦
华中科技大学同济医学院病理学系，卫生部呼吸系疾病重点实验室. 湖北, 武汉 430030
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W estern blot检测b-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b-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 b-连环素的细胞核内转位；锂能激活b-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。
关键词： 糖原合成酶激酶3; b-连环素; 鳞状分化; 气道上皮细胞; 猪
Role of glycogen synthase kinase 3 in squamous differentiation of pig airway epithelial cells: A primary study
CHEN Wen-Shu, WU Ren-Liang*, TIAN Dan, WANG Xi
Department of Pathology, Tongji Medical College, Huazhong University of Science and Technology
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 in morphology of cells was monitored under microscopy, and expression of b-catenin, phosphorylated GSK3 and involucrin, a squamous differentiation 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 the expression 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 observed after the elevation of squamous differentiation markers. Taken together, these data suggest that GSK3 is possibly involved in squamous differentiation of pig airway epithelial cells.
Key words: Glycogen synthase kinase 3;β-catenin;Squamous differentiation;Airway epithelial cell;Pig
陈文书, 吴人亮, 田丹, 王曦 . 糖原合成酶激酶3在猪气道上皮细胞鳞状分化中作用的初步探讨[J]. 生理学报 2005; 57 (4): .
CHEN Wen-Shu, WU Ren-Liang*, TIAN Dan, WANG Xi . Role of glycogen synthase kinase 3 in squamous differentiation of pig airway epithelial cells: A primary study. Acta Physiol Sin 2005; 57 (4): (in Chinese with English abstract).