蜂毒素调节人胃癌细胞线粒体相关蛋白表达  

Melittin regulates mitochondrial protein expression in human gastric cancer cells

作　　者：陶文华[1] 孔桂美[1] 卜平[1] 王继军[1] 胡茂志 张强[1] 

机构地区：[1]扬州大学医学院,江苏省扬州市225001 [2]扬州大学检测中心,江苏省扬州市225001

出　　处：《世界华人消化杂志》2013年第15期1400-1404,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;Nos.81272537;30801497;江苏省高校自然科学基金资助项目;No.11KJB360010~~

摘　　要：目的:观察蜂毒素(melittin)诱导人胃腺癌细胞凋亡及其对线粒体相关蛋白表达的影响.方法:4μg/mL melittin诱导SGC-7901细胞不同时间(0、1、2、4h),观察细胞的形态学改变;免疫荧光法观察活性氧(reactive oxygen species,ROS)、线粒体膜电位(Jc-1)表达;IHC观察胃癌细胞线粒体释放蛋白细胞凋亡诱导因子(apoptosis inducing factor,AIF)、Smac/Diablo、细胞色素C(cytochrome C)、EndoG的表达情况.结果:melittin能明显诱导SGC-7901细胞凋亡;ROS实验显示,melittin组与对照组比较,细胞胞浆出现很强的绿色荧光;MPT实验显示,melittin组显示绿色荧光的细胞数占多数(P<0.05),而阴性对照组,均表现为红色荧光细胞,线粒体相关蛋白检测结果显示,melittin组AIF、cytochrome C、EndoG、Smac/Diablo表达率分别为15.99%±1.66%、42.73%±3.48%、62.34%±2.71%、28.58%±2.09%(P<0.05).结论:melittin可诱导SGC-7901细胞凋亡,线粒体凋亡相关蛋白可能参与并发挥了重要作用.AIM： To observe the effect of melittin on apoptosis and expression of mitochondrial proteins in human gastric cancer cells （SGC-7901） in vitro. METHODS： SGC-7901 cells were treated with 4 ~g/mL of melittin for different durations （0, 1, 2, or 4 h）. After treatment, cell morphological changes were observed under an inverted microscope. The levels of reactive oxygen species （ROS） and mitochondrial transmembrane potential （MTP） were detected by fluorescence microscopy. The expression of apoptosis inducing factor （AIF）, Smac/Diablo, cytochrome C, and EndoG was exarnined by immunohistochemistry.RESULTS： Melittin induced the apoptosis of SGC-7901 cells. ROS experiments showed that compared to the control group, more strong cy- toplasmic green fluorescence was observed in the melittin group. MPT experiments showed that the majority of cells in the melittin group exhib- ited green fluorescence （P 〈 0.05）, while the con- trol group showed red fluorescence. The positive rates of mitochondrial proteins AIF, cytochrome C, EndoG, and Smac/Diablo protein expression were 15.99% ＋ 1.66%, 44.48% ＋ 4.85%, 62.34% ＋ 2.71%, and 28.58% ＋ 2.09%, significantly different from those in the control group （all P 〈 0.05）. CONCLUSION： Melittin induces SGC-7901 cell apoptosis possibly by regulating the expression of mitochondrial apoptosis related proteins.

关 键 词：蜂毒素 人胃腺癌SGC-7901细胞 活性氧 线粒体跨膜电位 凋亡诱导因子 细胞色素C 核酸内切酶 SMAC/DIABLO 

分 类 号：R735.2[医药卫生—肿瘤]