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目的:通过酸感受离子通道(ASIC)1 a表达抑制的细胞模型,探讨在酸性条件下ASIC1 a对细胞生长增殖的影响。方法:以大鼠神经胶质瘤(C6)为模型细胞,利用RNA i技术建立ASIC1 a表达抑制的细胞模型。通过绘制细胞生长曲线、集落形成试验、MTT试验和乳酸脱氢酶(LDH)释放试验来观察不同pH条件下,ASIC1 a表达抑制细胞和阴性对照细胞生长增殖能力的变化。结果:RNA i技术可明显抑制C6细胞ASIC1 a蛋白的表达。在正常pH条件下,ASIC1 a表达缺失对C6细胞的生长增殖没有显著影响。但在不同酸性条件下,ASIC1 a表达抑制细胞与阴性对照细胞相比对酸的耐受性明显提高,表现为细胞损伤减轻,存活数目明显增多。结论:C6细胞中有ASIC1 a,ASIC1b,ASIC2 a,ASIC2b和ASIC3亚基的表达。抑制ASIC1 a的表达不影响正常pH条件下细胞的生长增殖,但能明显提高细胞对酸的耐受性,提示ASIC1 a可能是潜在的抗酸靶分子。
OBJECTIVE: To investigate the effect of ASIC1 a on cell growth and proliferation under acidic conditions through an inhibitory cell model of acid-sensing ion channel (ASIC) 1 a expression. METHODS: Rat glioma C6 cells were used as model cells, and the cell model of ASIC1 a expression inhibition was established by using RNAi technique. Cell growth curve, colony formation assay, MTT assay and lactate dehydrogenase (LDH) release assay were used to observe the changes of ASIC1 a expression-suppressive cells and negative control cells under different pH conditions. Results: The RNAi technique could significantly inhibit the expression of ASIC1α protein in C6 cells. Under normal pH conditions, the loss of ASIC1 a expression had no significant effect on the proliferation of C6 cells. However, under different acidic conditions, the ASIC1 a-expressing suppressor cells showed significantly increased tolerance to acid compared with negative control cells, showing a reduction in cell damage and a significant increase in the number of survivors. Conclusion: There are ASIC1 a, ASIC1b, ASIC2a, ASIC2b and ASIC3 subunits in C6 cells. Inhibition of ASIC1 a expression did not affect the growth of cells under normal pH conditions, but significantly increased the acid tolerance of cells, suggesting that ASIC1 a may be a potential acid-fast target.