青蒿琥酯增加人脑胶质瘤细胞CHG-5对β射线的敏感性  

Artesunate enhances radiosensitivity of human glioma cell line CHG-5 to β-ray

作　　者：董俊清[1] 赵妍妍[1] 姚琦[1] 蒋为薇[1] 李军[1] 李斌[1] 庞学利[2] 周红[1] 

机构地区：[1]第三军医大学医学检验系暨药学院药理学教研室,重庆400038 [2]第三军医大学西南医院肿瘤科,重庆400038

出　　处：《第三军医大学学报》2010年第6期588-592,共5页Journal of Third Military Medical University

摘　　要：目的从青蒿素及其衍生物中筛选出对人脑胶质瘤细胞CHG-5及U87具有较好放射增敏效果的药物,并初步探讨其放射增敏机制。方法①采用MTT法测定上述青蒿素衍生物对2株胶质瘤细胞的IC50,观察以20%IC50浓度的青蒿素衍生物联合β射线对CHG-5及U87的放射增敏效果,从中筛选出放射增敏效果最好的药物;②通过克隆形成法计算目标药物的放射增敏比;③划痕法观察它对β射线照射后CHG-5细胞迁移的影响;④检测胞内还原型GSH水平;⑤流式细胞术检测细胞凋亡及细胞周期分布情况。结果青蒿素(artemisinin,ART)、双氢青蒿素(dihydroartemisinin,DHA)、青蒿琥酯(artesunate,AS)对CHG-5的IC50分别为:1.47、0.20、0.29mmol/L;对U87,则分别为1.61、0.25、0.46mmol/L。以20%IC50为β射线的协同给药浓度,仅有AS可以同时抑制经β射线照射后的CHG-5及U87细胞的增殖。且0.06mmol/L的AS对CHG-5细胞的放射增敏比为1.25,该浓度的AS可以明显地抑制CHG-5细胞的迁移。GSH结果表明0.06mmol/LAS处理组较未处理组细胞内GSH水平下调(P<0.05),流式细胞术结果提示0.06mmol/LAS可以阻滞细胞周期于G1期。结论从3种ART衍生物中筛选出AS对CHG-5细胞具有放射增敏作用,机制可能与下调的胞内GSH水平及细胞周期阻滞于G1期有关。Objective To investigate the cytotoxicity of 3 artemisinin derivatives, artemisinin （ART）, dihydroartemisinin （DHA） and artesunate （AS） to 2 glioma cell lines CHG-5 and U87, to select a compound wbich have radiosensitization on both cell lines when synergized with β-ray. Methods Firstly, we employed MTT assay to measure the IC50 of 2 cell lines to ART and its derivatives. When synergized with β-ray, the radiosensitization of drugs with 20% IC50 and selected the optimal one were observed. Subsequently, sensitivity enhancement ratio （SER） was determined by colony-forming assay. Scratch test was used to observe the effect of objective drug on cell migration. Then, intraeellular reduced GSH was measured. Finally, flow cytometry was employed to assay the cell apoptosis and cell cycle distribution. Results For CHG-5, the IC50 of ART, DHA and AS was 1. 47, 0.20 and 0.29 retool/L, respectively. Correspondingly, the IC50 for U87 was 1.61, 0.25 and 0.46 mmol/L, respectively. Only AS suppressed the proliferation of both cell lines under the concentration of 20% IC50 when combined with β-ray. Furthermore, the SER of 0. 06 mmol/L AS for CHG-5 was 1.25. Meanwhile, 0.06 mmol/L AS significantly inhibited the cell migration. Compared with medium group, GSH production was reduced after the treatment of 0.06 mmol/L AS （P 〈 0.05）. Flow cytometry indicated that the cell cycle was arrested in G1 phase after 0.06 mmol/L AS treatment. Conclusion Our results indicate that AS from the 3 artemisinin derivatives enhances the radiosensitivity of human glioma cell line CHG- 5 to β-ray, which may be associated with the decreased intracellnlar GSH and G1 arrest.

关 键 词：脑胶质瘤 青蒿素 衍生物 IC50 放射增敏 

分 类 号：R739.41[医药卫生—肿瘤] R965[医药卫生—临床医学]