Effects of Methadone on the Toll-like Receptor 4 Expression in Human Non-Small Cell Lung Carcinoma A549 Cell Line Using<i> In-silico</i> and <i>In vitro</i> Techniques

Abstract

Background: In this study, the effects of methadone and naloxone on the expression of toll-like receptor 4 (TLR4) gene have been evaluated in human non-small cell lung carcinoma A549 cell line migration using in-silico and in vitro techniques.Materials and Methods: Lung cancer A549 cell cultures were stimulated for 24 h with methadone (5, 10, and 20 mu M) and naloxone (20 and 40 mu M) concentrations. The level of TLR4 expression was determined by the quantitative real-time polymerase chain reaction. Migration of the A549 cells was investigated after a 4-h incubation period with methadone using the Boyden Chamber assay.Results: Migration rate of the A549 cells treated with 5 (P < 0.05) and 20 (P < 0.01) mu M methadone was, respectively, increased and decreased with 20 mu M naloxone (P < 0.05). Furthermore, the TLR4 expression was enhanced with 5 (P < 0.05) and 20 (P < 0.01) mu M methadone and decreased with 20 (P < 0.05) and 40 mu M naloxone (P < 0.01). In addition, in silico docking analysis revealed docking of methadone to MD-2 and TLR4.Conclusion: According to the present DATA, methadone affects the TLR4 expression. It may however cause adverse consequences by increasing the TLR4 expression. Therefore, the useful analgesic properties of methadone should be separated from the unwanted TLR4-mediated side effects.