青蒿琥酯诱导肿瘤细胞凋亡与抑制存活蛋白表达有关  

Apoptosis of tumor cells induced by artesunate is associated with suppressing survivin expression

作　　者：李哲[1] 袁守军[2] 聂丽平[1] 田增月[2] 徐兰平[2] 韩昌明[2] 

机构地区：[1]辽宁师范大学生命科学学院 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《中国临床药理学与治疗学》2004年第6期607-611,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金资助项目 (№ 3 0 2 715 18)

摘　　要：目的 :探讨青蒿琥酯 (artesunate ,Art)诱导肿瘤细胞凋亡与存活蛋白 (survivinprotein)表达的关系。方法 :采用细胞荧光染色、流式细胞术、琼脂糖凝胶电泳法和测定细胞浆Caspase 3的活性等手段检测肿瘤细胞暴露于不同浓度的Art时 ,对肿瘤细胞凋亡的诱导作用 ;用RT PCR、WesternBlotting法 ,检测不同浓度的Art作用于肿瘤细胞时 ,对survivinmRNA和survivin蛋白表达的影响。结果 :HL6 0细胞暴露于Art时 ,呈现典型细胞凋亡特征 ,如 :胞核固缩、形成凋亡小体 ;凋亡细胞的比例呈浓度依赖性增高 ;琼脂糖电泳出现明显的“梯状”条带 ;细胞浆Caspase 3的活性呈浓度依赖性增高等。RT PCR检测表明 ,A5 49细胞暴露于Art 10和 5 0g·L-172h后 ,survivinmRNA的表达呈浓度依赖性降低 ,对照组、10和 5 0mg·L-1处理组的survivin条带和内标GAPDH条带灰度的比值分别为 1.74 5、0 .390和0 .0 2 3;WesternBlotting法也检测到Art抑制Survivin蛋白的表达。结论 :Art诱导肿瘤细胞发生凋亡 ,激活Caspase 3途径 。AIM: To study the relations between apoptosis of tumor cells induced by artesunate (Art) and expression of survivin. METHODS: Tumor cells were exposured to different concentrations of Art, apoptosis was examined by fluorescent staining, flow cytometry, agarose gel electrophoresis (AGE) and Caspase-3 activity assay. The expressions of survivin mRNA and survivin protein were assayed with RT-PCR and Western blotting after cells treated with Art. RESULTS: With Art treatment, HL60 cells showed typical apoptotic features, including chromatin condensation and apoptotic bodies (fluorescent staining), and apoptosis rate of tumor cells increasing in concentration-dependent manners (flow cytometry), DNA ladder (AGE), activity of Caspase-3 enhancing. Analysis of RT-PCR indicated that expression of survivin mRNA was reduced in concentration-dependent manners after A549 cells were exposured to 10 and 50 mg·L -1 Art for 72 h. Intensity ratios between survivin strip and GAPDH strip in control, 10 and 50 mg·L -1 Art groups were 1.745, 0.390 and 0.023, respectively. Western blotting detection was also indicated that Art inhibited expression of survivin protein. CONCLUSION: Tumor cells apoptosis induced by Art might be associated with suppressing survivin expression.

关 键 词：青蒿素 青蒿琥酯 肿瘤 凋亡 作用机理 存活蛋白 

分 类 号：R965.1[医药卫生—药理学] R966[医药卫生—药学]