Underlying mechanism of ASIC1a involved in acidosis-induced cytotoxicity in rat C6 glioma cells
Abstract
Aim: To investigate the underlying mechanism of acid-sensing ion channel (ASIC) 1a involved in the acidosis-induced cytotoxicity of rat C6 glioma cells.
Methods: The stable ASIC1a-silenced C6 cells built with the RNA interference technology were confirmed by RT-PCR and Western blot analysis. Intracellular calcium ([Ca2+]i) in both the wild-type rat C6 glioma cells and the ASIC1a-silenced C6 cells were analyzed before and after acid application/exposure with the calcium imaging experiment.
Results: The rapid extracellular pH drop induced the increase of [Ca2+]i in the wild-type C6 cells, but not in the ASIC1a-silenced C6 cells. During the prolonged acid exposure, [Ca2+]i was lower in the ASIC1a-silenced C6 cells than that in the control cells.
Conclusion: The resultant toxicity of [Ca2+]i might contribute to the acidosis-induced cytotoxicity.
Keywords:
Methods: The stable ASIC1a-silenced C6 cells built with the RNA interference technology were confirmed by RT-PCR and Western blot analysis. Intracellular calcium ([Ca2+]i) in both the wild-type rat C6 glioma cells and the ASIC1a-silenced C6 cells were analyzed before and after acid application/exposure with the calcium imaging experiment.
Results: The rapid extracellular pH drop induced the increase of [Ca2+]i in the wild-type C6 cells, but not in the ASIC1a-silenced C6 cells. During the prolonged acid exposure, [Ca2+]i was lower in the ASIC1a-silenced C6 cells than that in the control cells.
Conclusion: The resultant toxicity of [Ca2+]i might contribute to the acidosis-induced cytotoxicity.