蟾毒灵联合顺铂对骨肉瘤143B细胞凋亡的影响

The Effect of Bufalin Combined with Ciplatin on Apoptosis of Osteosarcoma 143B Cells

摘要: 目的：探讨蟾毒灵和顺铂对骨肉瘤(Osteosarcoma, OS) 143B细胞增殖和凋亡的影响及其机制。方法：选取OS-143B细胞为实验细胞，分为阴性对照、蟾毒灵、顺铂和蟾毒灵 + 顺铂联合组四组。MTT法检测细胞增殖；荧光显微镜观察细胞凋亡形态；流式细胞仪检测细胞周期和细胞凋亡；Western blot检测Bcl-2、Bax和Caspase-3蛋白表达。结果：蟾毒灵和顺铂均抑制OS-143B细胞增殖(P < 0.01)，细胞生长减缓，细胞生长阻滞S期、G2期(P < 0.05)；细胞凋亡率均显著高于对照组(P < 0.01)；蟾毒灵和顺铂组较对照组Bax，caspase-3表达显著增加(P < 0.01)，而Bcl-2表达显著减低(P < 0.01)，上述结果联合组较两单药组作用更显著(P < 0.01)。结论：蟾毒灵与顺铂均可抑制OS-143B细胞增殖，呈时间–浓度依赖性，细胞周期在S期、G₂期阻滞；蟾毒灵与顺铂均可促进OS-143B细胞凋亡，可能与上调Bax，caspase-3蛋白，下调Bcl-2蛋白的表达有关。联合组较单用二药对抑制OS细胞生长、促进其凋亡的作用显著增强。

Abstract: Objective: To investigate the effects and mechanism of bufalin and cisplatin on proliferation and apoptosis of osteosarcoma 143B cells. Methods: OS-143B cells were selected as experimental cells and divided into four groups: the negative control group, the bufalin group, the cisplatin group and the bufalin + cisplatin group. MTT method was used to detect cell proliferation, fluorescence microscope to observe the morphology of apoptosis, flow cytometry to detect cell cycle and apoptosis, and western blot to detect the expression of Bcl-2, Bax and Caspase-3. Results: Both bufalin and cisplatin inhibited the proliferation of OS-143B cells, slowed down the cell growth, blocked S phase and G2 phase, and the apoptosis rate was significantly higher than that in the control group (P < 0.01). The expression of Bax, caspase-3 in the bufalin and cisplatin group was significantly higher than that in the control group, while the expression of Bcl-2 in the bufalin and cisplatin group was significantly lower than that in the control group (P < 0.01). The effect of the bufalin and cisplatin group was more significant than that of the two single drug groups. Conclusions: Both bufalin and cisplatin can inhibit the proliferation of OS-143B cells in a time-concentration dependent manner, and the cell cycle is arrested in S phase and G2 phase, and both bufalin and cisplatin can promote the apoptosis of OS-143B cells, which may be related to the up-regulation of Bax, caspase-3 protein and down-regulation of Bcl-2 protein expression. The effect of the combination group on inhibiting the growth and promoting the apoptosis of OS cells was significantly stronger than that of the two drugs alone.