线粒体凋亡途径参与TRAIL诱导胃腺癌细胞凋亡  

The mitochondrial apoptotic pathway is involved in TRAIL-induced apoptosis of gastric adenocarcinoma

作　　者：吴萍[1] 陈思[1] 程文晋[1] 江蓓蕾[1] 张旭东[1] 张林杰[1] 

机构地区：[1]安徽医科大学免疫学教研室,合肥230032

出　　处：《安徽医科大学学报》2007年第6期599-603,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30572118);安徽省自然科学基金(编号:070413077)

摘　　要：目的研究TRAIL诱导胃腺癌细胞凋亡的途径。方法PI染色、流式细胞仪检测细胞凋亡;广谱caspase抑制剂(z-VAD-fmk)、caspase-8抑制剂(z-IETD-fmk)和caspase-9抑制剂(z-LEHD-fmk)分别与TRAIL共同作用后PI染色、流式细胞仪检测细胞凋亡率的变化;Western blot检测caspase-3的活化及PARP的裂解;JC-1染色、流式细胞仪分析线粒体膜电位的变化。结果TRAIL诱导胃腺癌细胞凋亡具有剂量和时间依赖性,BGC-823较SGC-7901对TRAIL诱导的凋亡更敏感,TRAIL(100μg/L)作用24h细胞凋亡率分别是59.9%、24.3%;3种caspase抑制剂都能很大程度上阻止TRAIL诱导胃腺癌细胞凋亡;caspase-3、PARP在TRAIL作用早期即活化、裂解,且BGC-823比SGC-7901发生得更快;线粒体膜电位随TRAIL处理时间延长而不断下降。结论TRAIL能诱导胃腺癌细胞凋亡,且这种凋亡依赖于caspases;除死亡受体途径外,线粒体途径也参与了凋亡信号通路。Objective To explore the apoptotic pathway induced by TRAIL in gastric adenocarcinoma cells. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. Effect of caspase inhibitors on TRAIL-induced apoptosis were carried out by pretreating the cells with pan-caspase inhibitor （zVAD- fmk）, the caspase-8 inhibitor（ z-IETD-fmk）, or the caspase-9 inhibitor（z-LEHD-fmk） before adding TRAIL. The activation of caspase-3 and PARP cleavage were conducted by Western blot analysis. The changes in mitochondrial membrane potential were measured by uptake of JC-1 using flow cytometry. Results TRAIL induced apoptosis of gastric adenocarcinoma cells in a dose- and time-dependent manner. It appeared that BGC-823 was more sensitve to TRAIL-induced apoptosis than SGC-7901. The rates of apoptotic cells were 59, 9% and 24. 3%, respectively, after treatment with TRAIL（ 100 ~g/L） for 24 h. Caspase inhibitors blocked TRAIL-induced apoptosis nearly, completely. Caspase-3 activation and PARP cleavage were detected early after exposure to TRAIL. The mitochondrial membrane potential decreased in response to TRAIL. Conclusion Induction of apoptosis of gastric adenocarcinoma cells by TRAIL is caspase-dependent. TRAIL induces both receptor-mediated and mitochondrial intrinsic pathways.

关 键 词：胃肿瘤 细胞凋亡 线粒体 

分 类 号：R735.2[医药卫生—肿瘤] R329.2[医药卫生—临床医学]