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• W566858133 abstract "Polyaminesputrescine, spermidine, and spermineare involved in the growth of breast cancer cells. A possible target of polyamine action is at the site of interaction of transcription factors with their response elements. NF-κB is a member of the rel family of transcription factors that regulate transcription of genes in the proliferative/anti-apoptotic pathways. We performed electrophoretic mobility shift assays to study the role of polyamines in NF-κB binding to NF-κB response elements (NREs), the consensus sequence of which is GGGGAATTCCCC. Using cellular extract from MCF-7 breast cancer cells, we found very little binding of NF-κB to NRE in the absence of polyamines. Addition of 1 mM spermidine or spermine caused a 4- and 6-fold increase in NF-κB−NRE binding, respectively. Putrescine induced a 2-fold increase in the binding at 2 mM concentration. Using antibody supershift assays, we identified the p50 subunit of NF-κB to be a major component in NF-κB−NRE complex formation in the presence of polyamines. However, the decreased intensity of the band corresponding to NF-κB−NRE complex in the presence of anti-p65, c-rel, relB and p52 antibodies suggested the participation of these subunits also. Spermine also stimulated NF-κB−NRE binding using cellular extracts from other breast cancer cell lines and a normal breast epithelial cell line. A differential effect of spermine analogues on NF-κB−NRE binding was observed, with spermine exerting the maximal effect. CD spectra of NRE containing oligonucleotides was asymmetric and distinct from that of a typical B-DNA CD spectrum. A concentration-dependent increase in Tm of the duplex NRE was seen in the presence of polyamines. In transient transfection experiments using an NF-κB driven secreted alkaline phosphatase (SEAP) reporter, spermine induced NF-κB activity by ∼2-fold as compared to controls. Spermine induced activation of NF-κB was also confirmed using an NF-κB−EGFP (enhanced green fluorescent protein) vector in transient transfections in which expression of the green fluorescent protein was visualized by fluorescence microscopy. These data show a gene regulatory function of polyamines involving enhanced binding of NF-κB to NRE and a possible mechanism for the action of polyamines in breast cancer cell proliferation." @default.
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• W566858133 date "1999-10-23" @default.
• W566858133 modified "2023-10-02" @default.
• W566858133 title "Activation of Nuclear Factor κB by Polyamines in Breast Cancer Cells" @default.
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• W566858133 doi "https://doi.org/10.1021/bi991291v" @default.
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