双氢青蒿素联合顺铂对人神经母细胞瘤细胞增殖及凋亡的影响

Effects of dihydroartemisinin combined with cisplatin on proliferation and apoptosis in human neuroblastoma cells

目的:为提高顺铂的抗肿瘤效果,检测双氢青蒿素(DHA)与顺铂(CDDP)联合应用时,DHA是否能增强顺铂对人神经母细胞瘤细胞的生长抑制作用。方法:人神经母细胞瘤细胞SH-SY5Y为实验细胞株,分别使用不同浓度的DHA和CDDP单独和同时处理对数期肿瘤细胞;CCK-8法测定细胞的增殖抑制率,流式细胞术检测细胞周期及凋亡率;采用Compusyn软件计算DHA与CDDP的联合指数(CI)。Western Blot实验检测细胞凋亡相关蛋白(Bax、Bcl-2、Cleaved-PARP、PARP、Cleaved-Caspase3、Caspase3)的变化情况。结果:DHA单独用药时药物IC_(50)值为5.794μg/mL,CDDP单独用药时药物IC_(50)值为4.326μg/mL,DHA与CDDP联合用药时药物IC_(50)值为3.253μg/mL,DHA能抑制人神经母细胞瘤细胞增殖并呈浓度依赖性(P<0.05),促进细胞凋亡(P<0.05),与CDDP联用后作用更显著,且两药小剂量联用时作用拮抗,大剂量联用时作用协同。结论:DHA体外增强顺铂对人神经母细胞瘤细胞的抗肿瘤效应,提高人神经母细胞瘤细胞对顺铂治疗的敏感性。

Objective:To improve the antitumor effect of cisplatin,to test whether dihydroartemisinin(DHA) can enhance the inhibitory effect of cisplatin(CDDP) on the growth of human neuroblastoma cells when combined DHA with and cisplatin.Methods:Human neuroblastoma cell line SH-SY5Y was used as experimental cell line.The logarithmic stage tumor cells were treated with different concentrations of DHA and CDDP separately and simultaneously.Cell proliferation inhibition rate was determined by CCK-8 method,and cell cycle and apoptosis rate were determined by flow cytometry.Compusyn software was used to calculate the combination index(CI) of DHA and CDDP.The changes of apoptosis-related proteins(Bax,Bcl-2,Cleaved-PARP,PARP,Cleaved-Caspase3,Caspase3) were detected by Western Blot.Results:The IC_(50) value of DHA alone was 5.794 μg/mL,that of CDDP alone was 4.326 μg/mL,and that of DHA combined with CDDP was 3.253 μg/mL.DHA inhibited the proliferation of human neuroblastoma cells in a concentration-dependent manner(P<0.05) and promoted apoptosis(P<0.05).DHA combined with CDDP had a more significant effect,and the two drugs had an antagonistic effect in low dose and a synergistic effect in high dose.Conclusion:DHA enhances the antitumor effect of cisplatin on human neuroblastoma cells in vitro,and enhances the sensitivity of human neuroblastoma cells to cisplatin therapy.