Kotelevets Larissa, van Hengel Jolanda, Bruyneel Erik, Mareel Marc, van Roy Frans, Chastre Eric
INSERM U410, Faculté de Médecine Bichat, Paris, France.
FASEB J. 2005 Jan;19(1):115-7. doi: 10.1096/fj.04-1942fje. Epub 2004 Nov 1.
We recently established the critical role of the lipid phosphatase activity of the PTEN tumor suppressor in stabilizing cell-cell contacts and suppressing invasiveness. To delineate the effector systems involved, we investigated the interaction of PTEN with E-cadherin junctional complexes in kidney and colonic epithelial cell lines. PTEN and the p85 regulatory subunit of phosphatidylinositol 3-OH kinase (PI3K) co-immunoprecipitated with E-cadherin and catenins. By using a yeast two-hybrid assay, we demonstrated that PTEN interacted indirectly with beta-catenin by binding the scaffolding protein MAGI-1b. This model was corroborated in various ways in mammalian cells. Ectopic expression of MAGI-1b potentiated the interaction of PTEN with junctional complexes, promoted E-cadherin-dependent cell-cell aggregation, and reverted the Src-induced invasiveness of kidney MDCKts-src cells. In this model, MAGI-1b slightly decreased the activity of AKT, a downstream effector of PI3K. By using dominant-negative and constitutively active AKT expression vectors, we demonstrated that this kinase was included in the pathways involved in Src-induced destabilization of junctional complexes and was necessary and sufficient to trigger invasiveness. We propose that the recruitment of PTEN at adherens junctions by MAGI-1b and the local down-regulation of phosphatidylinositol-3,4,5-trisphosphate pools and downstream effector systems at the site of cell-cell contacts are focal points for restraining both disruption of junctional complexes and induction of tumor cell invasion.
我们最近确定了抑癌基因PTEN的脂质磷酸酶活性在稳定细胞间接触和抑制侵袭性方面的关键作用。为了阐明所涉及的效应系统,我们研究了PTEN与肾和结肠上皮细胞系中E-钙黏蛋白连接复合物的相互作用。PTEN和磷脂酰肌醇3-羟基激酶(PI3K)的p85调节亚基与E-钙黏蛋白和连环蛋白共免疫沉淀。通过酵母双杂交试验,我们证明PTEN通过结合支架蛋白MAGI-1b与β-连环蛋白间接相互作用。该模型在哺乳动物细胞中以多种方式得到证实。MAGI-1b的异位表达增强了PTEN与连接复合物的相互作用,促进了E-钙黏蛋白依赖性细胞间聚集,并逆转了Src诱导的肾MDCKts-src细胞的侵袭性。在该模型中,MAGI-1b略微降低了PI3K下游效应器AKT的活性。通过使用显性负性和组成型活性AKT表达载体,我们证明该激酶参与了Src诱导的连接复合物不稳定所涉及的途径,并且对于触发侵袭性是必要且充分的。我们提出,MAGI-1b在黏着连接处募集PTEN以及在细胞间接触部位局部下调磷脂酰肌醇-3,4,5-三磷酸池和下游效应系统是抑制连接复合物破坏和肿瘤细胞侵袭诱导的关键点。