钾通道阻滞剂对人肺腺癌细胞增殖和凋亡的影响

目的研究电压依赖性延迟整流钾通道阻断剂4-氨基吡啶(4-aminopyridine,4-AP)、钙离子激活钾通道阻断剂四乙铵(tetraethylammonia,TEA)对人肺腺癌细胞(AGZY-83-a)的影响,并探讨人肺腺癌细胞株凋亡的机制.方法体外细胞培养法培养人肺腺癌细胞株,采用MTT法检测不同浓度4-AP、TEA对细胞增殖的影响.透射电镜、流式细胞术(FCM)检测细胞凋亡情况.比色法检测细胞内Caspase-3活性改变.结果不同浓度的4-AP、TEA作用于细胞48h后,与对照组相比各组吸光度值(A490)均明显降低(n=10,P＜0.001),并且呈浓度依赖关系.4-AP、TEA组的IC50为3.59mmol/L和3.97mmol/L.4-AP及高浓度的TEA使人肺腺癌细胞株细胞凋亡率增加.经4-AP、TEA处理48h的AGZY-83-a细胞内Caspase-3的A405较正常对照组有显著增加(n=6,P＜0.001).结论钾通道阻滞剂4-AP、TEA对AGZY-83-a细胞增殖有明显的抑制作用,抑制率与药物浓度正相关.诱导人AGZY-83-a细胞凋亡,4-AP和TEA诱导AGZY-83-a细胞凋亡的过程中细胞内Caspase-3活性增加.

Effect of potassium channel blocker on proliferation and apoptosis of human pulmonary adenocarcinoma cells

Objective To investigate the effect of voltage dependent K~ channel blocker(4-aminopyridine,4-AP) and calcium activated K~ channel blocker(tetraethylammonia,TEA)on human pulmonary adenocarcinoma AGZY-83-a cell line and to explore its mechanism.Methods The inhibitory rate of cultured AGZY-83-a cell treated with 4-AP,TEA at various concentration was observed with MTT assay.Apoptosis was detected with flow cytometry and electronic microscope.Cellular caspase-3 activities were observed with colorimetric method.Results 48h after AGZY-83-a cell treated with different concentration of 4-AP,TEA administration,the absorbancy(A_(490)) was obviously lower than the control group accordingly(n=10,P<0.001),showing a concentration-dependent manner.The IC_(50) of 4-AP,TEA on cells were 3.59mmol/L and 3.97mmol/L,respectively.The flow cytometry analysis showed that 4-AP and high concentration TEA increased the percentage of early phase apoptosis.The cellular caspase-3 A_(405) of AGZY-83-a cell treated with 4-AP,TEA was obviously higher than control group(n=6,P<0.001).Conclusion The proliferation of AGZY-83-a cell is inhibited by potassium channel blocker 4-AP,TEA in concentration-dependent manner.4-AP,TEA induce apoptosis of AGZY-83-a cell.Caspase-3 is actived when 4-AP,TEA induce apoptosis of AGZY-83-a cell.