新疆家蚕抗菌肽诱导人胃癌细胞AGS凋亡的研究  

Apoptosis of human gastric carcinoma AGS cells induced by cecropin XJ

作　　者：吴艳玲[1] 夏丽洁[1] 张富春[1] 

机构地区：[1]新疆生物资源基因工程重点实验室、新疆大学生命科学与技术学院,新疆乌鲁木齐830046

出　　处：《中国药理学通报》2015年第2期186-193,共8页Chinese Pharmacological Bulletin

基　　金：新疆维吾尔自治区高技术研究发展计划项目(No201110101)

摘　　要：目的探讨新疆家蚕抗菌肽（cecropinXJ）是否通过诱导人胃癌细胞AGS凋亡产生抗肿瘤的作用。方法选择0．01～1000mg·L^-1浓度范围内的cecropinXJ与人胃癌细胞AGS和人正常胃上皮细胞GES-1共培养24h，采用MTT法检测cecropinXJ对AGS细胞和GES-1细胞增殖的影响；透射电镜观察细胞超微结构变化；Hoechst染色观察细胞凋亡情况；流式细胞术检测细胞内活性氧和线粒体膜电位的变化；实时荧光定量PCR（qRT-PCR）和Westernblot检测Bax、Bcl-2、caspase-3以及细胞色素CmRNA和蛋白水平的表达变化。结果CecropinXJ在体外能明显抑制胃癌AGS细胞的增殖（P〈0．05），并具有浓度依赖性，IC90值为61．19mg·L^-1，但对GES-1细胞无明显的抑制增殖作用。经cecropinXJ处理24h后，AGS细胞核固缩，呈现典型细胞凋亡特征，同时细胞内活性氧增加，线粒体膜电位下降。qRT-PCR和Westernblot结果表明，cecropinXJ能够引起Bcl-2表达下调，Bax表达上调，促进细胞色素C的释放并活化caspase-3。CecropinXJ促进caspase-3活性呈剂量依赖，经caspase-3和caspase-9特异性抑制剂处理后可降低cecropinXJ介导的AGS细胞死亡率。结论CecropinXJ可通过下调Bcl-2表达，上调Bax表达和活化caspase-3诱导AGS细胞凋亡，是其抗肿瘤机制之一。Aim To investigate the apoptosis of human gastric carcinoma AGS cells induced by cecropin XJ.Methods Human gastric carcinoma AGS cells and human normal epithelial cells GES-1 were co-cultured with different concentrations of cecropin XJ ranging from0. 01 to 1 000 mg·L- 1for 24 h. MTT assay was used to observe the effects of cecropin XJ on the proliferation of AGS cells and GES-1 cells. The ultrastructural changes of the AGS cells were observed by transmission electron microscopy. Hoechst staining was used to detect cell apoptosis. The changes of intracellular reactive oxygen species( ROS) and mitochondrial potential were analysed by flow cytometery. The expression of Bax,Bcl-2,caspase-3 and cytochrome C in mRNA level was investigated by qRT-PCR. Western blot was used to determine the protein expression of Bax,Bcl-2,caspase-3 and cytochrome C. Results Cecropin XJ significantly suppressed the proliferation of AGS cells in vitro( P < 0. 05) in a dose-dependent manner,IC50= 61. 19 mg·L- 1,but had no inhibitive effects on the proliferation of GES-1 cells. After treatment for 24 h,cecropin XJ induced AGS cells nuclear condensation,and increased ROS production,disrupted mitochondrial integrity. The results of qRT-PCR and Western blot demonstrated cecropin XJ could up-regulate the expression of Bax and down-regulate the expression of Bcl-2,promote the release of cytochrome C and activate caspase-3. Meanwhile,cecropin XJ promoted caspase-3activity in a dose-dependent manner,and cell death ratio of AGS cells induced by cecropin XJ was significantly reduced by caspase-3 and caspase-9 specific inhibitors treatment. Conclusion Cecropin XJ can induce apoptosis of AGS cells by downregulating Bcl-2,upregulating Bax and activating caspase-3,which may be one of its anti-tumor mechanisms.

关 键 词：新疆家蚕抗菌肽(cecropinXJ) 胃癌 AGS细胞 细胞凋亡 Bcl-2 BAX CASPASE-3 抗肿瘤 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R735.202.2[医药卫生—基础医学]