Significance: We characterize a mechanism through which the polarity protein atypical PKCι controls invasion and matrix remodeling by tumor cells by regulating endosome-to-plasma membrane traffic of the membrane type 1-matrix metalloproteinase (MT1-MMP) in breast-cancer cells. Further analysis shows that atypical PKCι and MT1-MMP are co–up-regulated in hormone receptor-negative breast tumors in association with higher risk of metastasis. These findings provide previously unidentified avenues for the design of therapeutic interventions.

During their metastatic spread, cancer cells need to remodel the extracellular matrix in order to migrate through stromal compartments adjacent to the primary tumor. Dissemination of breast carcinoma cells is mediated by membrane type 1-matrix metalloproteinase (MT1-MMP/MMP14), the main invadopodial matrix degradative component. Here, we identify MT1-MMP as a novel interacting partner of dual-specificity LIM Kinase-1 and -2 (LIMK1/2) and provide several evidence for phosphorylation of tyrosine Y573 in the cytoplasmic domain of MT1-MMP by LIMK. Phosphorylation of Y573 influences association of F-actin binding protein cortactin to MT1-MMP-positive endosomes and invadopodia formation and matrix degradation. Moreover, we show that LIMK1 regulates cortactin association to MT1-MMP-positive endosomes, while LIMK2 controls invadopodia-associated cortactin. In turn, LIMK1 and LIMK2 are required for MT1-MMP-dependent matrix degradation and cell invasion in a three-dimensional type I collagen environment. This novel link between LIMK1/2 and MT1-MMP may have important consequences for therapeutic control of breast cancer cell invasion.

In migrating NRK cells, aPKCs control the dynamics of turnover of paxillin-containing focal adhesions (FA) determining migration rate. Using a proteomic approach (2-DIGE), dynein intermediate chain 2 (dyneinIC2) was identified as a protein phosphorylated inducibly during cell migration in a PKC regulated manner. By gene silencing and co-immunoprecipitation studies, we show that dyneinIC2 regulates the speed of cell migration dependent on its interaction with paxillin. This interaction is controlled by Serine 84 phosphorylation, which lies on the aPKC pathway. The evidence presented thus links aPKC control of migration to the dynein control of FA turnover through paxillin.

Cell competition is a tissue surveillance mechanism for eliminating unwanted cells, being indispensable in development, infection and tumourigenesis. Although studies have established the role of biochemical mechanisms in this process, due to challenges in measuring forces in these systems, how mechanical forces determine the competition outcome remains unclear. Here we report a form of cell competition that is regulated by differences in force transmission capabilities, selecting for cell types with stronger intercellular adhesion. Direct force measurements in ex vivo tissues and different cell lines reveal that there is an increased mechanical activity at the interface between two competing cell types, which can lead to large stress fluctuations resulting in upward forces and cell elimination. We show how a winning cell type endowed with a stronger intercellular adhesion exhibits higher resistance to elimination and benefiting from efficient force transmission to the neighbouring cells. This cell elimination mechanism could have broad implications for keeping the strong force transmission ability for maintaining tissue boundaries and cell invasion pathology.

Metastasis is the main cause of cancer-related deaths. How a single oncogenic cell evolves within highly organized epithelium is still unknown. Here, we found that the overexpression of the protein kinase atypical protein kinase C ι (aPKCi), an oncogene, triggers basally oriented epithelial cell extrusion in vivo as a potential mechanism for early breast tumor cell invasion. We found that cell segregation is the first step required for basal extrusion of luminal cells and identify aPKCi and vinculin as regulators of cell segregation. We propose that asymmetric vinculin levels at the junction between normal and aPKCi⁺ cells trigger an increase in tension at these cell junctions. Moreover, we show that aPKCi⁺ cells acquire promigratory features, including increased vinculin levels and vinculin dynamics at the cell–substratum contacts. Overall, this study shows that a balance between cell contractility and cell–cell adhesion is crucial for promoting basally oriented cell extrusion, a mechanism for early breast cancer cell invasion. Significance This study shows that an oncogenic mammary epithelial cell surrounded by normal cells can extrude basally in vivo and invade surrounding tissues without formation of a primary tumor. Here, we show that overexpression of the key polarity protein atypical protein kinase C ι (aPKCi) is sufficient for triggering basally oriented epithelial cell extrusion and early cell invasion into the mammary gland stroma. Moreover, we highlight the importance of the difference between the mechanical properties of aPKCi-overexpressing cells and those of the normal surrounding cells associated with the decrease of vinculin at the cell junction, which triggers cell segregation, the first step toward promoting and controlling the direction of cell extrusion.