Journal: International Journal of Molecular Sciences
Article Title: Oxaliplatin Causes Transient Changes in TRPM8 Channel Activity
doi: 10.3390/ijms22094962
Figure Lengend Snippet: Phosphatidyl 4,5-bisphosphate concentrations are altered after oxaliplatin treatment. ( A ) Relative PIP 2 (phosphatidylinositol 4,5-bisphosphate) concentrations of untransfected HEK cells, empty-vector-transfected and TRPM8-transfected HEK cells treated with vehicle (saline) or 10 µM oxaliplatin for 24 h. Data represent the means ± SEM from n = 10–12 dishes per condition. * p < 0.05; *** p < 0.001; one-way ANOVA and Dunnett´s multiple comparison test. ( B ) Relative PIP 2 concentrations of untransfected HEK cells, empty-vector-transfected and TRPM8-transfected HEK cells that were treated with vehicle (saline), 10 µM oxaliplatin ±10 µM of the PLC inhibitor D609 for 5 h. Data represent the means ± SEM from n = 4–6 dishes per condition. *** p < 0.001; one-way ANOVA and Dunnett´s multiple comparison test. ( C ) Schematic overview of the proposed mechanisms leading to TRPM8 channel desensitization after acute oxaliplatin treatment. TRPM8 channel activation causes PLC pathway activation. In turn, PLC hydrolyzes PIP 2 to IP 3 (inositol 1,4,5-triphosphate) and DAG (1,2-diacylglycerol). PIP 2 -depletion is responsible for TRPM8 channel desensitization after oxaliplatin treatment.
Article Snippet: For PI (4,5) P2 ELISA measurement analysis, (r)TRPM8-transfected or empty-vector-transfected and untransfected HEK cells were treated with 10 μM oxaliplatin for 24 h. For investigating the contribution of the PLC pathway to the PIP 2 amount, (r)TRPM8-transfected or empty-vector-transfected and untransfected HEK cells were treated with 10 μM oxaliplatin ±10 μM of the PLC inhibitor D609 (Tocris) for 5 h.
Techniques: Plasmid Preparation, Transfection, Saline, Comparison, Activation Assay
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