Cooperative interactions between p53 and NFκB enhance cell plasticity

// Alessandra Bisio 1 , Judit Zamborszky 1, 3 , Sara Zaccara 1 , Mattia Lion 1, 4 , Toma Tebaldi 2 , Vasundhara Sharma 1 , Ivan Raimondi 1 , Federica Alessandrini 1 , Yari Ciribilli 1 , Alberto Inga 1 1 Laboratory of Transcriptional Networks, Centre for Integrative Biology, CIBIO, University of Trento, Trento, 38123, Italy 2 Laboratory of Translational Genomics, Centre for Integrative Biology, CIBIO, University of Trento, Trento, 38123, Italy 3 Institute of Enzymology, Research Centre for Natural Sciences, Budapest, Hungary 4 Department of Genetics, Massachusetts General Hospital, Boston, MA, USA Correspondence to: Yari Ciribilli, e-mail: ciribilli@science.unitn.it Alberto Inga, e-mail: inga@science.unitn.it Keywords: p53, NFkB, chemotherapy, doxorubicin, TNFα, EMT, synergy, breast cancer Received: June 25, 2014      Accepted: October 01, 2014      Published: October 21, 2014 ABSTRACT The p53 and NFκB sequence-specific transcription factors play crucial roles in cell proliferation and survival with critical, even if typically opposite, effects on cancer progression. To investigate a possible crosstalk between p53 and NFκB driven by chemotherapy-induced responses in the context of an inflammatory microenvironment, we performed a proof of concept study using MCF7 cells. Transcriptome analyses upon single or combined treatments with doxorubicin (Doxo, 1.5μM) and the NFκB inducer TNF-alpha (TNFα, 5ng/ml) revealed 432 up-regulated (log 2 FC> 2), and 390 repressed genes (log 2 FC< −2) for the Doxo+TNFα treatment. 239 up-regulated and 161 repressed genes were synergistically regulated by the double treatment. Annotation and pathway analyses of Doxo+TNFα selectively up-regulated genes indicated strong enrichment for cell migration terms. A panel of genes was examined by qPCR coupled to p53 activation by Doxo, 5-Fluoruracil and Nutlin-3a, or to p53 or NFκB inhibition. Transcriptome data were confirmed for 12 of 15 selected genes and seven (PLK3, LAMP3, ETV7, UNC5B, NTN1, DUSP5, SNAI1) were synergistically up-regulated after Doxo+TNFα and dependent both on p53 and NFκB. Migration assays consistently showed an increase in motility for MCF7 cells upon Doxo+TNFα. A signature of 29 Doxo+TNFα highly synergistic genes exhibited prognostic value for luminal breast cancer patients, with adverse outcome correlating with higher relative expression. We propose that the crosstalk between p53 and NFκB can lead to the activation of specific gene expression programs that may impact on cancer phenotypes and potentially modify the efficacy of cancer therapy.