三氧化二砷通过线粒体依赖性通路诱导喉癌细胞株及其耐药株细胞凋亡  

Arsenic Trioxide Induced Cell Apoptosis by Mitochondria Dependent Pathway in KB and KBv200 Cells

作　　者：李艳芳[1] 王旭东[1] 梁永钜[1] 陈黎明[1] 丁岩[1] 黄民[2] 符立梧[1] 

机构地区：[1]中山大学肿瘤防治中心抗癌药物室 [2]中山大学药学院,广东广州510089

出　　处：《癌症》2004年第6期631-634,共4页Chinese Journal of Cancer

基　　金：国家自然科学基金(No.30371659);广东省自然科学基金(No.2003C30313)~~

摘　　要：背景与目的:三氧化二砷(arsenictrioxide,As2O3)已作为治疗实体瘤的新药应用于临床,但其作用机理仍不清楚。本研究拟探讨As2O3通过线粒体依赖性凋亡通路介导喉癌细胞及其耐药株细胞凋亡的作用。方法:采用MTT法测定As2O3对喉癌细胞KB及其耐药细胞KBv200的细胞毒作用;用AnnexinVFITC染色法检测凋亡早期细胞;细胞线粒体跨膜电位(ΔΨm)用DiOC6标记,流式细胞仪检测。结果:As2O3对KB及KBv200细胞的生长有明显的抑制作用,IC50分别为(0.22±0.02)μg/ml和(0.20±0.01)μg/ml。用AnnexinVFITC染色检测到As2O3呈时间依赖性介导细胞凋亡,2.5μg/mlAs2O3处理24h、48h,KB细胞凋亡率分别为(20.2±3.1)%和(52.2±11.0)%;KBv200细胞凋亡率分别为(15.8±1.3)%和(36.4±5.9)%。2.5μg/mlAs2O3作用于KB与KBv200细胞,ΔΨm降低呈时间依赖性。结论:ΔΨm降低在As2O3诱导喉癌细胞凋亡过程中起着重要的作用。BACKGROUND &OBJECTIVE: Arsenic trioxide (As2O3) is a new drug used to treat the patients with solid tumor,but the mechanism is still unclear. This study was designed to investigate the effect of mitochondrial dependent pathway in apoptosis induced by arsenic trioxide in multidrug resistant KBv200 cells an d their parental sensitive KB cells. METHODS: The cytotoxic effect of As2O3 on K B and KBv200 cells was measured by MTT assay. KB and KBv200 cells were treated r espectively with As2O3 for 12, 24, 48 hours. Cell apoptosis was determined by An nexin V FITC staining. Mitochondrial membrane potential was labeled by DiOC6 and examined by flow cytometry. RESULTS: Arsenic trioxide showed the inhibition of KB and KBv200 cells proliferation in vitro. The IC50s of As2O3 to KB and KBv200 cells were (0.22±0.02)靏/ml and (0.20±0.01)靏/ml, respectively. As2O3 in duced cell apoptosis in time-dependent manner. The apoptosis rates were 20.2% ±3.1%and 52.2%±11.0%for KB cells and 15.8%±1.3%and 36.4%±5.9%for KBv2 00 cells under 2.5 靏/ml As2O3 treating 24 h and 48 h, respectively. The levels of mitochondrial membrane potential were concentration-dependently decreased a fter treating with arsenic trioxide for 12, 24, and 48 hours. CONCLUSION: The de crease of mitochondrial membrane potential maybe plays an important role in apop tosis induced by As2O3.

关 键 词：三氧化二砷 MTT 细胞凋亡 线粒体跨膜电位 

分 类 号：R739.6[医药卫生—肿瘤]