TLRs and Bcl-2 family proteins in neutrophils of oral cavity cancer patients.

Abstract

Human neutrophils (PMNs), the cells engaged in the early phase of anti-tumor response, express TLR2 and TLR6 that can modulate the Bcl-2 family proteins, regulating the intrinsic apoptotic pathway in these cells. The expression of TLRs and Bcl-2 family is controlled by means of activating the transcriptional signaling pathways that involve the p38 MAP kinase. As previously described, PMNs from cancer patients exert accelerated apoptosis associated with decreased expression of anti-apoptotic Mcl-1 protein. In the present study we have been interested in establishing the involvement of TLR2 and TLR6, and p38 MAP kinase in the Mcl-1-modulated apoptosis in PMNs of oral cavity cancer patients. The expression of these proteins in neutrophils and autologous peripheral blood mononuclear cells (PBMCs) was analyzed by Western blot, the intensity of apoptosis was estimated by flow cytometry, caspase-9 activity by colorimetric assay, and the cytochrome c concentration by ELISA. The simultaneous decreased expression of examined TLRs receptors and Mcl-1 protein, associated with the acceleration of PMNs apoptosis, suggests that this process in PMNs controlled by Mcl-1 is dependent on the TLR2 and TLR6 signalling. Impaired TLRs expression can lead to insufficient activation of p38PAPK, resulting in low expression of antiapoptotic Mcl-1 protein responsible for shortened lifespan of the examined PMNs.