Li Shaowei, Braverman Richard, Li Hongzhen, Vass William C, Lowy Douglas R, DeClue Jeffrey E
Laboratory of Cellular Oncology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Mol Carcinog. 2003 Jun;37(2):98-109. doi: 10.1002/mc.10125.
We investigated the effects of overexpression of the tuberous sclerosis-1 and -2 (TSC1/2) gene products (hamartin and tuberin, respectively) in the human kidney epithelial cell line 293 with an inducible expression system. As we had observed previously in fibroblasts, 293 cells overexpressing hamartin and/or tuberin grew more slowly in vitro. However, here we also observed that the 293 overexpressing cells underwent a dramatic morphological change in which groups of cells formed compact clusters. The overexpressing cells also displayed decreased dissociation and increased reaggregation in vitro. These changes were found to be associated with an increased level of E-cadherin, which is known to regulate cell-cell interactions in epithelial cells, and of its binding partner beta-catenin. Consistent with the role of E-cadherin in these effects, we found that the observed changes in 293 cell morphology, dissociation, and adhesion were calcium-dependent, and were reproduced by overexpression of E-cadherin. In contrast, overexpression of TSC1 in rat embryo fibroblasts, which lack E-cadherin, failed to elicit the same changes as in 293 cells. We conclude that the hamartin/tuberin complex exerted a direct effect on the morphology and adhesive properties of 293 cells through regulation of the level and/or activity of cellular E-cadherin/beta-catenin.
我们利用诱导表达系统研究了结节性硬化症1和2(TSC1/2)基因产物(分别为错构瘤蛋白和结节蛋白)在人肾上皮细胞系293中的过表达效应。正如我们之前在成纤维细胞中观察到的那样,过表达错构瘤蛋白和/或结节蛋白的293细胞在体外生长得更慢。然而,在这里我们还观察到,过表达的293细胞发生了显著的形态变化,细胞群形成了紧密的簇。过表达的细胞在体外还表现出解离减少和重新聚集增加。这些变化被发现与E-钙黏蛋白及其结合伴侣β-连环蛋白水平的增加有关,已知E-钙黏蛋白可调节上皮细胞中的细胞间相互作用。与E-钙黏蛋白在这些效应中的作用一致,我们发现293细胞形态、解离和黏附中观察到的变化是钙依赖性的,并且通过E-钙黏蛋白的过表达得以重现。相比之下,在缺乏E-钙黏蛋白的大鼠胚胎成纤维细胞中过表达TSC1未能引发与293细胞相同的变化。我们得出结论,错构瘤蛋白/结节蛋白复合物通过调节细胞E-钙黏蛋白/β-连环蛋白的水平和/或活性,对293细胞的形态和黏附特性产生直接影响。