艾迪协同卡铂高效杀伤骨肉瘤细胞的实验研究

Killing effect of osteosarcoma cells induced by combination of AiDi and carboplatin

目的:研究中药制剂艾迪对骨肉瘤化疗的增效作用及其相关机制。方法:将艾迪与卡铂单独及联合应用于骨肉瘤OS-732细胞,用MTT法检测细胞抑制率,流式细胞仪定量分析凋亡细胞所占比例,倒置相差显微镜及荧光显微镜下观察凋亡细胞形态改变,免疫细胞化学技术分析凋亡相关蛋白Fas表达。结果:艾迪与卡铂对骨肉瘤细胞都有剂量依赖性杀伤作用,艾迪浓度达到25μl/ml时,细胞抑制率为38.47%,再增加艾迪浓度抑制率不再明显改变,但如与1μg/ml卡铂合用将使细胞抑制率进一步提升为59.53%(P<0.01)。细胞超微结构观察及凋亡率测定也显示,艾迪与卡铂联用可诱导更多骨肉瘤细胞凋亡,且联用时骨肉瘤Fas表达明显提升(P<0.01)。结论:艾迪与小剂量卡铂合用和单用卡铂相比,可更高效杀伤骨肉瘤细胞,此作用与上调Fas表达诱导骨肉瘤细胞凋亡有关。

Objective：To study the killing effect of osteosarcoma cells induced by combination of AiDi and carboplatin and related mechanism.Methods：AiDi and carboplatin were applied on the osteosarcoma cells respectively or jointly and inhibition rate was measured by MTT assay.The celluar ultrastructure were observed with inverted phase contrast microscope and fluorescence microscope.The apoptotic rate was analyzed with flow cytometry（FCM） Imunocytochemistry was used to examine the expression of Fas on OS-732 cells.Results：Both AiDi and carboplatin had the killing effect on OS-732 cell lines depending on the drug doze.The inhibition rate was 38.47% when 25μl / ml AiDi was used on OS-732 cell lines and the inhibition rate did not rise significantly with more AiDi application,if combined with 1μg / ml carboplatin however,inhibition rate was 59.53% which was significantly higher than that of 25μl /ml AiDi individually（P 0.01）.Changes of celluar ultrastructure and apoptotic rate also indicated apoptosis-inducing effect of joint application was much stronger than those of respective application,in addition,up-regulated expression of Fas and induced apoptosis were also observed along with the killing effect of OS-732 cell lines（P 0.01）.Conclusion：Combination of AiDi and small dose of carboplatin has stronger killing effect on OS-732 cell lines compared with carboplatin alone which may be initiated by up-regulated Fas expression and induced apoptosis.