Inactivation of Focal Adhesion Kinase FAK Rapidly Abrogates Keratinocyte Entry in Mitosis via Rho‐Associated Kinase, Resulting in Squamous Differentiation

Cell adhesion tightly controls cell proliferation and homeostasis in stratified epithelia by mechanisms that remain unclear. Focal adhesion kinase (FAK) transduces cell adhesion signals, is frequently deregulated in epithelial cancer, and it has been associated with proliferation and resistance to treatments. The mechanisms by which FAK controls the epithelial cell cycle are still intriguing. We previously unraveled a mitosis-differentiation checkpoint that is the limiting factor in the keratinocyte cell cycle. To investigate whether FAK plays a role in this checkpoint, we inactivated the protein in normal human oral keratinocytes by specific shRNAs or by the specific inhibitor defactinib. Inactivation of FAK very rapidly and strikingly blocked entry into mitosis and triggered a differentiation response. This response was independent of DNA damage. Tumor suppressor P53 was induced shortly after inhibition of FAK, while mitotic Cyclin B was not translocated into the nucleus. Human epidermal N-TERT cells that were synchronized in prometaphase failed to execute mitosis. Concomitant inhibition of FAK-downstream Rho-associated kinase (Rock) rescued mitotic progression. The results unveil a rapid Rock-dependent mitosis switch upon inactivation of FAK, inducing terminal differentiation, pointing at a mitotic automatic mechanism of epithelia to suppress suprabasal proliferation of precancerous cells.