Toscani Andrés Martín, Sampayo Rocío G, Barabas Federico Martín, Fuentes Federico, Simian Marina, Coluccio Leskow Federico
Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Biológica, Buenos Aires, Argentina.
CONICET, Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), Buenos Aires, Argentina.
PLoS One. 2017 Mar 17;12(3):e0174230. doi: 10.1371/journal.pone.0174230. eCollection 2017.
ErbB2 is a member of the ErbB family of tyrosine kinase receptors that plays a major role in breast cancer progression. Located at the plasma membrane, ErbB2 forms large clusters in spite of the presence of growth factors. Beta1 integrin, membrane receptor of extracellular matrix proteins, regulates adhesion, migration and invasiveness of breast cancer cells. Physical interaction between beta1 integrin and ErbB2 has been suggested although published data are contradictory. The aim of the present work was to study the interaction between ErbB2 and beta1 integrin in different scenarios of expression and activation. We determined that beta1 integrin and ErbB2 colocalization is dependent on the expression level of both receptors exclusively in adherent cells. In suspension cells, lack of focal adhesions leave integrins free to diffuse on the plasma membrane and interact with ErbB2 even at low expression levels of both receptors. In adherent cells, high expression of beta1 integrin leaves unbound receptors outside focal complexes that diffuse within the plasma membrane and interact with ErbB2 membrane domains. Superresolution imaging showed the existence of two distinct populations of ErbB2: a major population located in large clusters and a minor population outside these structures. Upon ErbB2 overexpression, receptors outside large clusters can freely diffuse at the membrane and interact with integrins. These results reveal how expression levels of beta1 integrin and ErbB2 determine their frequency of colocalization and show that extracellular matrix proteins shape membrane clusters distribution, regulating ErbB2 and beta1 integrin activity in breast cancer cells.
ErbB2是酪氨酸激酶受体ErbB家族的成员,在乳腺癌进展中起主要作用。尽管存在生长因子,ErbB2仍位于质膜上并形成大的簇。β1整合素是细胞外基质蛋白的膜受体,可调节乳腺癌细胞的粘附、迁移和侵袭性。尽管已发表的数据相互矛盾,但有人提出β1整合素与ErbB2之间存在物理相互作用。本研究的目的是研究在不同表达和激活情况下ErbB2与β1整合素之间的相互作用。我们确定,β1整合素与ErbB2的共定位仅在贴壁细胞中取决于两种受体的表达水平。在悬浮细胞中,缺乏粘着斑使整合素可以在质膜上自由扩散并与ErbB2相互作用,即使两种受体的表达水平都很低。在贴壁细胞中,β1整合素的高表达使未结合的受体留在粘着斑复合物之外,这些受体在质膜内扩散并与ErbB2膜结构域相互作用。超分辨率成像显示存在两种不同的ErbB2群体:主要群体位于大簇中,次要群体位于这些结构之外。在ErbB2过表达时,大簇外的受体可以在膜上自由扩散并与整合素相互作用。这些结果揭示了β1整合素和ErbB2的表达水平如何决定它们的共定位频率,并表明细胞外基质蛋白塑造了膜簇的分布,调节了乳腺癌细胞中ErbB2和β1整合素的活性。