Effect Of TLR4 Activation On The Cell Proliferation And Cytokine Secretion Of Oral Squamous Cell Carcinoma

ObjectiveThe purpose of this project is to investigate the expression of Toll-like receptors(TLRs)in Tca8113 cells and the effect of toll-like receptor(TLR)4 activation on the cell proliferation and cytokine secretion of Tca8113 cells.Methods1.TLRs expression was detected by quantitative realtime PCR and flowcytometry in Tca8113 cells.2.Quantitative realtime PCR and flowcytometry were applied to detect the change of TLR4 m RNA and protein levels,after 10 ?g/ml LPS deal with Tca8113 cells.3.MTT assay was applied to determine the cell proliferation,after different concentrations of LPS deal with Tca8113 cells 24 h,48 h,72 h.4.NF-?B was detected by the Western blot,after different concentrations of LPS deal with Tca8113 cells 30 min,2 h,16 h.5.Interleukin(IL)-6 and IL-8 secretion were examined by ELISA,after 10 ?g/ml LPS deal with Tca8113 cells 16 h.Results1.All tested TLRs including TLR2,3,4,5,7,and 9 were expressed in Tca8113 cells.TLR 3,5 were weakly expressed,whereas the expression of TLR2,4,7,9 was relatively strong,2.The m RNA and protein levels of TLR4 increased,after 10 ?g/ml LPS deal with Tca8113 cells.3.MTT assay results show that: 1 ?g/ml and 10 ?g/ml LPS significantly induced the proliferation of Tca8113 cells(P<0.05).When cells were treated in the presence of 2 ?g/ml anti-TLR-4 monoclonal antibody for 30 min and then treated in 10 ?g/ml LPS for 72 h,proliferation rate was significantly inhibited(P<0.05).4.LPS can activate NF-?B signal pathway in Tca8113 cells.5.MTT assay results show that: IL-6 production was increased by the treatment of 1 ?g/ml and 10 ?g/ml LPS(P<0.05),IL-8 production was increased only by the treatment of 10 ?g/ml LPS(P<0.05).Conclusions1.LPS can increase the expression of TLR4 in Tca8113 cells.2.Activation of TLR4 can promote the proliferation of Tca8113 cells.3.Activation of TLR4 can activate NF-?B signaling pathway,promote the secretion of IL-6,IL-8 from Tca8113 cells.