手霉素通过反应性氧基诱导甲状腺未分化癌KAT-4细胞凋亡  

Manumycin-induced apoptosis via reactive oxygen species in KAT-4 anaplastic thyroid cancer cells

作　　者：李劲高[1] 佘妙容[2] 黄湖辉[1] 宛霞[1] 

机构地区：[1]中山大学附属第二医院肾内科,广州510120 [2]广东省人民医院血液科,广州510080

出　　处：《中华普通外科学文献（电子版）》2008年第4期23-25,共3页Chinese Archives of General Surgery(Electronic Edition)

基　　金：广东省自然科学基金项目(7001196);广东省医学科研基金项目(A2006020);广东省中医药局基金(2007180)

摘　　要：目的研究手霉素对甲状腺未分化癌KAT-4细胞的作用,探讨其作用机制。方法使用人的甲状腺未分化癌KAT-4细胞,MTT细胞毒性测定评价手霉素对KAT-4细胞的作用,使用annexin v和一氧化氮(nitric oxide,NO)染料标记细胞,应用流式细胞仪术检测细胞内NO生成和细胞凋亡;细胞内超氧阴离子通过二氢乙啶(dihydroethidium,DHE)测定;采用荧光法测定谷胱甘肽(glutathione,GSH)。结果手霉素降低KAT-4细胞活性,且呈剂量依赖性。手霉素诱导KAT-4细胞凋亡和反应性氧基(reactive oxygen species,ROS)产生包括NO和超氧阴离子增加和GSH降低。手霉素诱导细胞凋亡与ROS的增加有关,N-乙酰基-L-半胱氨酸(N-acetyl-L-cysteine,NAC)可抑制ROS产生,保护KAT-4细胞逃避手霉素的细胞毒作用和避免手霉素诱导的凋亡。结论细胞内ROS的产生对手霉素的细胞毒作用起非常重要的作用。手霉素通过增加ROS产生诱导甲状腺癌细胞凋亡。Objective To investigate the effects of manumycin on KAT-4 anaplastic thyroid cancer cell line, and to explore the mechanism. Methods Human anaplastic thyroid cancer cells KAT-4 were used. The cytotoxicity was analyzed by MTT assay. Apoptosis and cellular nitric oxide were detected by flow cytometry using annexin v and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by DHE. GSH was assayed by fluorescent monochlorobimane. Results Manumycin resulted in viability decrease in a dose-dependent manner. Manumycin induced apoptosis and ROS generation including nitric oxide and superoxide anions. Manumycin reduced intracellular glutathione. Manumycin-induced apoptosis correlated with increase in ROS. Quenching of ROS with N-acetyl-L-cysteine protected leukemia cells from the cytotoxicity of manumycin and prevented apoptosis induction by manumycin. Conclusion Cellular ROS generation plays an important role in the cytotoxic effect of manumycin. Manumycin induces apoptosis through ROS generation in KAT-4 cells.

关 键 词：甲状腺未分化癌 手霉素 凋亡 反应性氧基 

分 类 号：R736.1[医药卫生—肿瘤] R285.5[医药卫生—临床医学]