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Cytoplasmic mutant p53 increases Bcl-2 expression in estrogen receptor-positive breast cancer cells

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Abstract

The Bcl-2 gene is positively regulated by estrogen (E2) primarily through E2-response elements in the coding region and a putative p53 negative regulatory element (NRE) containing a short upstream open reading frame (uORF). The ability of mutant p53 to repress or induce Bcl-2 expression is controversial. In this study E2-receptor positive (ER+)/wild-type p53 MCF-7cells were transfected with p53Δ291, which lacks a nuclear localization signal or a DNA binding domain mutant, p53(173L). Both p53 mutants but especially p53Δ291 increased Bcl-2 protein expression from a CMV-NRE-Bcl-2 cDNA construct in an NRE-position/orientation independent manner as well as from a 1.7 kb Bcl-2 promoter reporter gene. Bcl-2 protein expression prevented the p53Δ291-mediated increase in Bcl-2 promoter activity although immunoprecipitation demonstrated that only a small proportion of the wild-type p53 but not p53Δ91 protein interacts with Bcl-2. Unless levels of ectopically expressed mutant p53 were extremely high, stable expression of mutant p53 in MCF-7 cells moderately increased Bcl-2 protein levels. Expression of mutant p53 did not alter E2 regulation of Bcl-2, however, mutation of the uORF prevented regulation by both mutant p53 and E2. Adenovirus-mediated overexpression of WT p53 strongly reduced Bcl-2 expression in ER−/mut p53 MDA-MB-231 cells. Taken together these data support the position that mutant p53 behaves in a dominant “positive” manner relieving repression by WT p53 or another Bcl-2 transcriptional inhibitor in a manner independent of nuclear translocation.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, University of Ottawa, 451 Smyth Road, Ottawa, Ontario, Canada, K1H 8M5

    M. A. Christine Pratt, Dawn White, Neena Kushwaha, Emma Tibbo & Min Ying Niu

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  1. M. A. Christine Pratt
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  2. Dawn White
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  3. Neena Kushwaha
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  4. Emma Tibbo
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  5. Min Ying Niu
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Correspondence to M. A. Christine Pratt.

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Supported by a grant from the National Cancer Institute of Canada/Canadian Institutes of Health Research Breast Cancer Initiative to M. A. C. P.

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Pratt, M.A.C., White, D., Kushwaha, N. et al. Cytoplasmic mutant p53 increases Bcl-2 expression in estrogen receptor-positive breast cancer cells. Apoptosis 12, 657–669 (2007). https://doi.org/10.1007/s10495-006-0023-y

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