The relationship between members of the canonical NF-kB pathway, components of tumour microenvironment and survival in patients with invasive ductal breast cancer

The aim of the present study was to examine the relationship between tumour NF-kB activation, tumour microenvironment including local inflammatory response (LIR) and cancer-specific survival in patients with operable ductal breast cancer. Immunohistochemistry (tissue microarray of 376 patients) and western blotting (MCF7 and MDA-MB-231 breast cancer cells) was performed to assess expression of key members of the canonical NF-kB pathway (inhibitory kappa B kinase (IKK beta) and phosphorylated p65 Ser-536 (p-p65)). Following silencing of IKK beta, cell viability and apoptosis was assessed in both MCF7 and MDA-MB-231 cell lines. P-p65 was associated with cancer-specific survival (CSS) (nuclear P=0.042 and total P=0.025). High total p-p65 was associated with increase grade tumour grade (P=0.010), ER positivity (P=0.023), molecular subtype (P=0.005), lower Klintrup-Makinen grade (P=0.013) and decreased CD138 count (P=0.032). On multivariate analysis, total p-p65 expression independently associated with poorer CSS (P=0.020). In vitro work demonstrated that the canonical NF-kB pathway was inducible by exposure to TNFa in ER-positive MCF7 cells and to a lesser extent in ER-negative MDAMB-231 cells. Reduction of IKK beta expression by siRNA transfection increased levels of apoptosis and reduced cell viability in both MCF7 (P=<0.001, P=<0.001, respectively) and MDA-MB-231 cells (P=>0.001, P=0.002, respectively). This is consistent with the hypothesis that canonical IKK beta-NF-kB signalling drives tumour survival. These results suggest that activation of the canonical NF-kB pathway is an important determinant of poor outcome in patients with invasive ductal breast cancer.