Nasopharynx epithelial cells are constantly exposed to both commensal and pathogenic micro-organisms. After being stimulated by some microbial, the nasopharynx epithelial cells secrete a lot of inflammatory factors that lead to the inflammation of nasopharynx and further result in the nasopharyngeal carcinoma and other disorders. LPS is the most important commensal or pathogenic bacteria constituents. It is related to the pathogenesis of a variety of disease. It has been shown that the 5-8F cells could bind with the FITC-labeling LPS for its expression of CD14, TLR4 and MD2 with flow cytometry and RT-PCR assay. With immunofluoresense, Western-blot and luciferase reporter system assay, it is indicated that LPS activated the TLR4 signaling pathway in 5-8F cells. Phospho-NFκB p65 expression was increased and entered into the nuclear in 5-8F cells with LPS inducement. Furthermore, after LPS stimulation, TNF-α promoter activity and the relevant amount of TNF-α being produced were increased in 5-8F cells. In conclusion, 5-8F cells expressed CD14, TLR4 and MD2 that are crucial for LPS binding. When nasopharnyx epithelial cells were induced by LPS, they did respond to LPS via TLR4 signaling pathways and activated NFκB signaling pathway, which can further lead to nasopharnyx inflammation and other nasopharnyx disorders.