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目的:探讨氯通道在三氧化二砷(arsenic trioxide,As_2O_3)诱导人鼻咽癌CNE-2Z细胞凋亡中的作用。方法:采用流式细胞术检测CNE-2Z细胞经As_2O_3作用24及48 h的凋亡率;应用膜片钳技术记录As_2O_3激活的细胞膜电流,并分析其电流特性;采用流式细胞术检测氯通道阻断剂DIDS对As_2O_3诱导的CNE-2Z细胞凋亡的抑制作用。结果:(1)5μmol/L As_2O_3能够时间依赖性地诱导的CNE-2Z细胞凋亡;(2)CNE-2Z细胞外灌流含5μmol/L As_2O_3的等渗溶液能够激活一个具有外向整流特征的电流,激活的电流无明显的时间及电压依赖性失活,翻转电位接近氯平衡电位;(3)As_2O_3激活的电流能够被氯通道阻断剂DIDS和NPPB完全抑制,细胞外灌流47%的高渗溶液能够完全抑制As_2O_3激活的电流;(4)氯通道阻断剂DIDS能够抑制As_2O_3诱导的CNE-2Z细胞凋亡。结论:As_2O_3可以激活CNE-2Z细胞的容积敏感性氯通道。氯通道在As_2O_3诱导的CNE-2Z凋亡中发挥重要作用。
Objective: To investigate the role of chloride channel in the apoptosis of human nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As 2 O 3). Methods: The apoptotic rate of CNE-2Z cells treated with As_2O_3 at 24 and 48 h was measured by flow cytometry. Cell membrane current activated by As_2O_3 was recorded by patch-clamp technique. The current characteristics of CNE-2Z cells were analyzed by flow cytometry. Inhibitory effect of blocking agent DIDS on apoptosis of CNE-2Z cells induced by As 2 O 3. Results: (1) Apoptosis of CNE-2Z cells induced by 5μmol / L As_2O_3 can be time-dependently induced by 5μmol / L As_2O_3. (2) The CNE-2Z extracellular perfusion solution containing 5μmol / L As_2O_3 can activate an outward rectifying current (3) The currents activated by As 2 O 3 were completely inhibited by chloride channel blockers DIDS and NPPB, and the extracellular perfusion of 47% of the hypertonic medium Solution can completely inhibit the current of As 2 O 3 activation; (4) DIDS chloride channel blocker can inhibit As 2 O 3 induced CNE-2Z cell apoptosis. Conclusion: As 2 O 3 can activate volume-sensitive chloride channels in CNE-2Z cells. Chloride channels play an important role in the apoptosis of CNE-2Z induced by As 2 O 3.