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Aim:To study the role of acid-sensing ion channel (ASIC) 1a in the cell death andapoptosis induced by extracellular acid in C6 glioma cells.Methods:The stableASIC1a-silenced C6 cell line,built with RNA interference technology,were con-firmed by RT-PCR and Western blot analysis.The cell viability following acidexposure was analyzed with lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.The apoptoticcells dyed with Annexin-V and propidium iodide were measured with a flowcytometer,while the changes of cell cycle were also assayed.Results:Thedownregulation of ASIC1a proteins by stable transfection of short hairpin RNAdecreased the cell death percentage and increased cell viability following acidexposure with LDH and the MTT assay.The rate of apoptosis was lower in theASIC1a-silenced cell line than that in the wild-type C6 cell line.The percentage ofsub-G_0 cells was lower in the ASIC1a-silenced C6 cells than that in the wild-typecells.Conclusion:Extracellular acid induced cell death and apoptosis via ASIC1amechanisms in the C6 glioma cells.
Aim: To study the role of acid-sensing ion channel (ASIC) 1a in the cell death and apoptosis induced by extracellular acid in C6 glioma cells. Methods: The stable ASIC 1a-silenced C6 cell line, built with RNA interference technology, were con-firmed by RT-PCR and Western blot analysis. The cell viability following acidexposure was analyzed with lactate dehydrogenase (LDH) and 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay. apoptotic cells dyed with Annexin-V and propidium iodide were measured with a flowcytometer, while the changes of cell cycle were also assayed. Results: The downregulation of ASIC1a proteins by stable transfection of short hairpin RNA decreased the cell death percentage and increased cell viability following acidexposure with LDH and the MTT assay. The rate of apoptosis was lower in the ASIC1a-silenced cell line than that in the wild-type C6 cell line. The percentage of sub-G_0 cells was lower in the ASIC1a-silenced C6 cells than that in the wild-type cells. Conclusion: Extracellular acid induced cell death and apoptosis via ASIC mechanisms in the C6 glioma cells.