Breast cancer cell proliferation is inhibited by BAD: Regulation of cyclin D1

Romaine Fernando, James S. Foster, Amber Bible, Anders Ström, Richard G. Pestell, Mahadev Rao, Arnold Saxton, Joon Baek Seung, Kiyoshi Yamaguchi, Robert Donnell, Maria Cekanova, Jay Wimalasena

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Abstract

Recent investigations suggest that functions of the proapoptotic BCL2 family members, including BAD, are not limited to regulation of apoptosis. Here we demonstrate that BAD inhibits G1 to S phase transition in MCF7 breast cancer cells independent of apoptosis. BAD overexpression inhibited G1 transit and cell growth as well as cyclin D1 expression. Inhibition of cyclin D1 expression was mediated through inhibition of transcription activated by AP1. Chromatin immunoprecipitation assays indicated that BAD is localized at the 12-O-tetradecanoylphorbol-13-acetate-response element (TRE) and cAMP-response element (CRE) in the cyclin D1 promoter. This was shown to reflect direct binding interactions of BAD with c-Jun, and this interaction inhibited the activity of AP1 complexes at TRE. BAD did not interact with phosphorylated forms of c-Jun. Our data suggest that inhibitory TRE/CRE-c-Jun-BAD complexes are present at the cyclin D1 promoter in quiescent cells. Estrogen stimulation displaced BAD from TRE/CRE elements in MCF7 cells, whereas BAD overexpression inhibited estrogen-induced cyclin D1 synthesis and cell proliferation. Inhibition of endogenous BAD in MCF7 cells markedly increased the proliferative fraction and DNA synthesis, activated Cdks, and increased cyclin D1 protein levels. This action of BAD required serine residues Ser75 and Ser99. Both phosphorylated and unphosphorylated forms of BAD localized to the nuclei of human breast epithelial cells. Thus, we demonstrate a novel role for BAD in cell cycle regulation dependent upon its phosphorylation state and independent of the BAD/BCL2 interaction and apoptosis.

Original languageEnglish
Pages (from-to)28864-28873
Number of pages10
JournalJournal of Biological Chemistry
Volume282
Issue number39
DOIs
Publication statusPublished - 28-09-2007

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All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Fernando, R., Foster, J. S., Bible, A., Ström, A., Pestell, R. G., Rao, M., ... Wimalasena, J. (2007). Breast cancer cell proliferation is inhibited by BAD: Regulation of cyclin D1. Journal of Biological Chemistry, 282(39), 28864-28873. https://doi.org/10.1074/jbc.M700785200