Dachshund inhibits oncogene-induced breast cancer cellular migration and invasion through suppression of interleukin-8

Proteomic analysis of DACH1-regulated chemokines. (A) Transwell migration assay of MDA-MB-231 cells treated with conditioned media derived from either vehicle control or ponasterone A-treated cells. (B) Cytokine array analysis of supernatant derived from MDA-MB-231 cells treated with either vehicle or ponasterone A to induce DACH1 identified a subset of cytokines differentially regulated by DACH1. Relative abundance is shown, comparing DACH1 expressing cells (PonA) with control. (C) Transwell migration assays of MDA-MB-231 cells treated with cytokines as indicated, including interleukin-8 or control IgG at 5 ng/ml. Migration of cells is shown as cell number per field with data as mean ± SEM for n > 5 separate experiments. (D) 3D collagen invasion assay of MCF10A-Ras cells expressing control (GFP) or DACH1 in the presence of IL-8 or vehicle. The mean depth of migration is shown quantified as mean ± SEM. (E) Cellular migration of MEFs derived from Dach1 WT or knockout mice, using time lapse video microscopy. Dach1^−/− (KO) MEFs display enhanced cellular migratory velocity and distance (P < 0.01). (F) The number of lung metastasis identified in nude mice injected with Met-1 cells transduced with pLRT-DACH1, treated with vehicle or Doxycycline DACH1 or immunoneutralizing antibody to CRCL1/KC. *, P < 0.01.