Nicotinic receptors modulate antitumor therapy response in triple negative breast cancer cells

Abstract

Triple negative breast cancer is more aggressive than other breast cancer subtypes and constitutes a public health problem worldwide since it has high morbidity and mortality due to the lack of defined therapeutic targets. Resistance to chemotherapy complicates the evolution of these patients. Several authors have highlighted the participation of nicotinic acetylcholine receptors (nAChR) in the modulation of conventional chemotherapy treatment in lung, head and neck, oral and nasal cavity, and pancreatic cancers. However, in smoking cancer patients, the action of nicotine on nAChR expressed in the breast or other organs near the tumor during chemotherapy treatment is less known.AIMTo investigate the effect of nicotine on paclitaxel treatment and the signaling pathways involved in human breast MDA-MB231 tumor cells.METHODSCells were treated with paclitaxel alone or in combination with nicotine, administered for one or three 48 h cycles. The effect of the addition of nicotine (in a concentration similar to that found in smokers? blood) on the treatment with paclitaxel (in a therapeutic concentration) was determined by using the 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The signaling mediators involved in this effect were determined by using selective inhibitors. We also investigated nAChR expression, and ATP ?binding cassette? G2 drug transporter (ABCG2) expression and its modulation by the different treatments by Western blot. The effect of the treatments on apoptosis induction was determined by flow cytometry using annexin-V and 7AAD markers. RESULTSOur results confirmed that the treatment with paclitaxel reduced MDA-MB231 cell viability in a concentration-dependent manner and that the presence of nicotine reversed the cytotoxic effect induced by paclitaxel by involving the expression of functional α7 and α9 nAChRs in these cells. The action of nicotine on paclitaxel treatment was linked to the modulation of the protein kinase C, mitogen-activated protein kinase, extracellular signal-regulated kinase, and NF-B signaling pathways, and to an up-regulation of ABCG2 protein expression. We also detected that nicotine significantly reduced the increase in cell apoptosis induced by paclitaxel treatment. Moreover, we observed that the presence of nicotine reduced the efficacy of paclitaxel treatment administered in three cycles to MDA-MB231 tumor cells. CONCLUSIONOur findings point to nAChRs as responsible for the decrease in the chemotherapeutic effect of paclitaxel in triple negative tumors. Thus, they should be considered as targets in smoking patients.