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目的探讨Wnt/β-catenin信号通路活化对人表皮细胞表型变化的影响。方法差速贴壁法分离人成熟表皮细胞。并将细胞分为对照组和诱导组,诱导组又分为氯化锂(LiCl,20mmol/L)诱导组和GSK-3β抑制子(15μmol/L)诱导组,诱导6d后观察细胞的形态变化,并采用Westernblot检测表皮细胞内β-catenin的表达水平,免疫组织化学法检测表皮细胞表面标志性蛋白CK10、CK14、CK19和β1整合素的表达变化。结果人表皮细胞内β-catenin的表达在被LiCl(2.15±0.54)和GSK-3β(2.58±0.65)抑制子诱导后分别比对照组(0.71±0.23)增加了2倍和2.5倍(P<0.01)。而且诱导后细胞体积变小变圆,细胞核、细胞质比例变大。免疫组织化学检测结果表明,诱导前表皮细胞高表达CK10,部分细胞表达CK14,CK19和β1整合素表达阴性;诱导后已无CK10阳性细胞存在,有部分CK14阳性细胞,而且CK19和β1整合素的表达呈强阳性。结论 Wnt/β-catenin通路活化可引起人成熟表皮细胞的去分化,即Wnt/β-catenin通路可能是调控人表皮细胞去分化的关键环节。
Objective To investigate the effect of Wnt / β-catenin signaling pathway on the phenotype of human epidermal cells. Methods Differential adherence method was used to separate human mature epidermal cells. The cells were divided into the control group and the induction group. The induced group was further divided into groups of LiCl (20mmol / L) and GSK-3β (15μmol / L) Western blot was used to detect the expression of β-catenin in epidermal cells. The expression of epidermal cell surface markers CK10, CK14, CK19 and β1 integrin were detected by immunohistochemistry. Results The expression of β-catenin in human epidermal cells increased 2-fold and 2.5-fold compared with the control group (0.71 ± 0.23) after being induced by LiCl (2.15 ± 0.54) and GSK-3β (2.58 ± 0.65) 0.01). After the induction, the cell size becomes smaller and round, and the ratio of nucleus and cytoplasm becomes larger. The results of immunohistochemistry showed that CK10 was highly expressed in the epidermal cells before induction, and some cells expressed negative CK14, CK19 and β1 integrin; no CK10 positive cells existed after the induction, and some CK14 positive cells were found, and CK19 and β1 integrin The expression was strongly positive. Conclusion Activation of Wnt / β-catenin pathway leads to the dedifferentiation of human mature epidermal cells. Wnt / β-catenin pathway may be the key point regulating the dedifferentiation of human epidermal cells.