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Updated: Jul 13, 2026

Three-Dimensional (3D) Tumor Spheroid Invasion Assay
Published on: May 1, 2015
Proliferation-associated Brn-3b transcription factor can activate cyclin D1 expression in neuroblastoma and breast
V S Budhram-Mahadeo1, S Irshad, S Bowen
1Medical Molecular Biology Unit, Institute of Child Health, University College London, London, UK. v.mahadeo@ich.ucl.ac.uk
Abstract:
Brn-3b transcription factor enhances proliferation of neuroblastoma (NB) and breast cancer cell lines in vitro and increases the rate and size of in vivo tumour growth, whereas reducing Brn-3b slows growth, both in vitro and in vivo. Brn-3b is elevated in >65% of breast cancer biopsies, and here we demonstrate that Brn-3b is also elevated in NB tumours. We show a significant correlation between Brn-3b and cyclin D1 (CD1) in breast cancers and NB tumours and cell lines. Brn-3b directly transactivates the CD1 promoter in co-transfection experiments, whereas electrophoretic mobility shift assay and chromatin immunoprecipitation assays demonstrate that Brn-3b protein binds to an octamer sequence located in the proximal CD1 promoter. Site-directed mutagenesis of this sequence resulted in loss of transactivation of the CD1 promoter by Brn-3b. Thus, Brn-3b may act to alter growth properties of breast cancer and NB cells by enhancing CD1 expression in these cells.
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