Glucocorticoid down-regulation of fascin protein expression is required for the steroid-induced formation of tight junctions and cell-cell interactions in rat mammary epithelial tumor cells

Glucocorticoid hormones, which are physiological regulators of mammary epit helium development, induce the formation of tight junctions in rat Con8 mam mary epithelial tumor cells. We have discovered that, as part of this proce ss, the synthetic glucocorticoid dexamethasone strongly and reversibly down -regulated the expression of fascin, an actin-bundling protein that also in teracts with the adherens junction component beta-catenin, Ectopic constitu tive expression of full-length mouse fascin containing a Myc epitope tag (M yc-fascin) in Con8 cells inhibited the dexamethasone stimulation of transep ithelial electrical resistance, disrupted the induced localization of the t ight junction protein occludin and the adherens junction protein beta-caten in to the cell periphery, and prevented the rearrangement of the actin cyto skeleton, Ectopic expression of either the carboxyl-terminal 213 amino acid s of fascin, which includes the actin and beta-catenin-binding sites, or th e amino-terminal 313 amino acids of fascin failed to disrupt the glucocorti coid induction of tight junction formation. Mammary tumor cells expressing the full-length Myc-fascin remained generally glucocorticoid responsive and displayed no changes in the levels or protein-protein interactions of junc tional proteins or the amount of cytoskeletal associated actin filaments, H owever, a cell aggregation assay demonstrated that the expression of Myc-fa scin abrogated the dexamethasone induction of cell-cell adhesion. Our resul ts implicate the down-regulation of fascin as a key intermediate step that directly links glucocorticoid receptor signaling to the coordinate control of junctional complex formation and cell-cell interactions in mammary tumor epithelial cells.