Increased Cell Migration and Plasticity in Nrf2 Deficient Cancer Cell Lines

Abstract

Nuclear Factor (erythroid-derived 2)-like 2 (Nrf2) expression is deregulated in many cancers. Genetic and biochemical approaches coupled with functional assays in cultured cells were used to explore the consequence of Nrf2 repression. Nrf2 suppression by Keap1-directed ubiquitylation or expression of independent shRNA/siRNA sequences enhanced cellular ROS, Smad-dependent tumor cell motility, and growth in soft agar. Loss of Nrf2 was accompanied by concomitant Smad linker region/C-terminus phosphorylation, induction of the E-Cadherin transcriptional repressor Slug, and suppression of the cell-cell adhesion protein E-Cadherin. Ectopic expression of wildtype Nrf2, but not dominant negative Nrf2, suppressed the activity of a synthetic TGF-β1 responsive CAGA-directed luciferase reporter. shRNA knock-down of Nrf2 enhanced the activity of the synthetic CAGA-reporter, as well as the expression of the endogenous Smad target gene plasminogen activator inhibitor-1. Finally, we found that Nrf2/Smad3/Smad4 formed an immunoprecipitable nuclear complex. Thus, loss of Nrf2 increased R-Smad phosphorylation and R-Smad signaling, supporting the hypothesis that loss of Nrf2 in an oncogenic context-dependent manner can enhance cellular plasticity and motility, in part by using TGF-β/Smad signaling.