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目的探讨比较华蟾素(cinobufagin)协同顺铂(cisplatin)对人骨肉瘤细胞株MG-63的杀伤作用。方法体外培养骨肉瘤细胞株MG-63细胞,取对数生长期细胞进行实验;采用MTT比色法测定MG-63细胞的生长抑制作用并计算细胞抑制率;流式细胞仪检测骨肉瘤细胞的凋亡率和细胞周期,观察细胞凋亡率的变化情况和药物对细胞周期的抑制作用;原位末端标记法(TUNEL)观察骨肉瘤细胞的凋亡情况并检测细胞凋亡率;相差显微镜观察骨肉瘤细胞生长状况及形态学上的改变。结果华蟾素和顺铂均对骨肉瘤细胞有生长抑制作用,华蟾素组、顺铂组及华蟾素+顺铂组在药物浓度不同时对骨肉瘤细胞的生长抑制作用各不相同,在药物浓度低于80μg/ml时,各实验组药物对细胞的生长抑制作用随药物浓度的增加而上升;当药物浓度达到80μg/ml时,华蟾素组和顺铂组药物对骨肉瘤细胞的生长抑制作用不再增加,但华蟾素+顺铂组药物对细胞的生长抑制作用在药物浓度达到160μg/ml时,抑制作用仍强于其浓度为80μg/ml时。各实验组在药物浓度相同时华蟾素+顺铂对骨肉瘤细胞的生长抑制作用明显强于单独应用华蟾素或者顺铂。结论华蟾素+顺铂对骨肉瘤MG-63细胞的联合杀伤作用明显强于单独应用顺铂或者华蟾素;在抑制骨肉瘤MG-63细胞生长方面,华蟾索协同顺铂对细胞的杀伤作用效果显著,并且具有时间和浓度依赖性。
Objective To investigate the killing effect of cinobufagin and cisplatin on human osteosarcoma cell line MG-63. Methods The osteosarcoma MG-63 cells were cultured in vitro and the cells in logarithmic growth phase were cultured. The growth inhibition of MG-63 cells was measured by MTT colorimetric assay and the inhibition rate of cells was calculated. The expression of osteosarcoma cells Apoptotic rate and cell cycle were observed to observe the changes of cell apoptosis rate and drug’s inhibition on cell cycle. The apoptosis of osteosarcoma cells was observed by TUNEL and the apoptosis rate was detected by phase contrast microscopy Osteosarcoma cell growth and morphological changes. Results Both cinobufotalin and cisplatin had inhibitory effects on the growth of osteosarcoma cells. The inhibitory effects of cinobufacini, cisplatin and cinobufotalin plus cisplatin on the growth of osteosarcoma cells were different at different drug concentrations, When the drug concentration was lower than 80μg / ml, the inhibitory effect of each experimental group on the cell growth increased with the increase of the drug concentration; when the drug concentration reached 80μg / ml, the cinobufacini and cisplatin group drugs on osteosarcoma cells The growth inhibitory effect no longer increased, but the cinobufotalin + cisplatin group inhibited the growth of cells when the drug concentration reached 160μg / ml, the inhibitory effect was still stronger than its concentration of 80μg / ml. The inhibitory effects of cinobufotalin and cisplatin on osteosarcoma cells were significantly stronger than those of cinobufacine or cisplatin alone in the experimental groups at the same drug concentration. Conclusion Cinobufacini combined with cisplatin is more effective in killing osteosarcoma MG-63 cells than cisplatin or cinobufacine alone. In the inhibition of the growth of osteosarcoma MG-63 cells, The killing effect is remarkable and time-dependent and concentration-dependent.