The in vivo antitumoral effects of lipopolysaccharide against glioblastoma multiforme are mediated in part by toll-like receptor 4

OBJECTIVE: Toll-like receptor 4 (Tlr-4) mediates many biological effects of lipopolysaccharide (LPS), which has antitumoral effects on glioblastoma both in vivo and in vitro. However, the precise role of Tlr-4 in these antitumoral effects remains unknown. METHODS: The role of Tlr-4 in the antitumoral effect of LPS on glioblastomas was assessed in wild-type BALB/c mice and in Tlr-4 knockout (KO) BALB/c mice. Mice were implanted with DBT glioblastoma cells intracranially or subcutaneously, were treated with intratumoral LPS, and were assessed by histopathological examination for degrees of tumor progression and inflammation. Flow cytometry and Western blotting with antibodies to the Tlr-4 receptor and flow cytometry to the related CD14 moiety were performed to quantitate the expression levels of these two receptors by glioblastoma cells. RESULTS: For subcutaneous tumors, LPS caused near complete tumor elimination in wild-type mice, but only a 50% reduction in Tlr-4 KO mice. For mice implanted with intracranial glioblastomas, LPS increased survival times modestly in wild-type mice, but showed no benefit in the Tlr-4 KO mice. There were no histological differences among wild-type and Tlr-4 KO mice, except for tumor size. In both models, an early neutrophilic and later macrophage-rich inflammatory infiltrate were seen after LPS administration. Quantitative flow cytometry and Western blotting showed no Tlr-4 receptor or CD14 expression in murine and human glioblastoma cells in vitro, and Western blotting suggested that Tlr-4 effects are mediated by nontumoral elements such as microglia and inflammatory cells. CONCLUSION: LPS-induced antitumoral effects on glioblastoma multiforme are mediated, in part, by the Tlr-4 receptor. Further understanding of this process may lead to novel treatment strategies for this uniformly fatal disease.