目的：初步探讨蟾毒灵对人肠癌HCT8细胞凋亡及钙离子代谢的影响及其机制。方法：采用细胞活性CCK-8法检测不同浓度的蟾毒灵以及相同浓度不同时间孵育后其对人肠癌HCT8细胞增殖的影响；运用Annexin V/PI双染凋亡试剂盒检测不同浓度的蟾毒灵对细胞凋亡的影响情况；采用Ca2+荧光探针Fluo-3 AM孵育人结肠癌HCT8细胞并标记其细胞内的Ca2+，检测不同浓度的蟾毒灵对细胞内Ca2+的影响。结果：CCK-8法结果显示，蟾毒灵对人结肠癌细胞的抑制作用呈明显的剂量以及时间依赖性；流式凋亡结果表明，蟾毒灵能诱导HCT8细胞发生凋亡，且呈一个剂量依赖性；根据Ca2+荧光探针标记胞内钙离子的结果显示，越高浓度的蟾毒灵作用下，细胞内的钙离子浓度越高。结论：蟾毒灵能诱导人结肠癌HCT8细胞凋亡，可能与细胞内的Ca2+代谢相关。
Objective：To study the influence and mechanism of bufalin on apoptosis and calcium metabolism in human colorectal cancer cell HCT 8. Methods：We used the method of CCK-8 to detect the proliferation of HCT 8 treated with bufalin of different concentrations at 24 hours or the same one at different time. And then , the apoptosis of the cell with different concentrations was detected by Annexin V / PI double staining apoptosis kit. Finally, we investigated the effect of different concentrations of bufalin on intracellular Ca2 + by using Ca2+ fluorescence probe Fluo-3 AM to mark its intracellular Ca2+. Results：The inhibition of human colorectal cancer cell HCT 8 with bufalin showed dose and time dependent manner obviously by the method of CCK-8.
The result of apoptosis with flow cytometry illustrated that bufalin can induce apoptosis in the cell HCT 8, and represented a dose-dependent manner. According to the results of the intracellular Ca2+ labeled by calcium fluorescent probe calcium, there was a higher concentration of intracellular calcium when the cell treated at a higher concentration of bufalin. Conclusion：Bufalin can induce apoptosis in the cell of HCT 8, and it may be related to the metabolism of intracellular Ca2+.
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