毒毛旋花子阿洛糖苷诱导胃腺癌细胞SGC-7901凋亡作用及机制研究  

Apoptosis of Gastric Adenocarcinoma Cells Type SGC- 7901 Induced by Strophalloside and Its Mechanism

作　　者：张雪娇[1] 梅文莉[2] 蔡彩虹[2] 董文化[2] 姜北[3] 黄风迎[4] 戴好富[2] 

机构地区：[1]大理学院,云南省大理市671099 [2]中国热带农业科学院热带生物技术研究所 [3]大理学院药物研究所 [4]海南医学院海南省热带病重点实验室

出　　处：《中国全科医学》2015年第15期1743-1747,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(30860345);国家科技支撑计划课题资助项目(2013BAI11B04);公益性行业(农业)科研专项(201303117)

摘　　要：目的探讨毒毛旋花子阿洛糖苷诱导胃腺癌细胞SGC-7901凋亡作用及其机制。方法将人胃腺癌细胞SGC-7901分为对照组、0.125μg/ml组、0.250μg/ml组、0.500μg/ml组、1.000μg/ml组,四甲基偶氮唑盐微量酶反应比色法(MTT)检测毒毛旋花子阿洛糖苷对胃腺癌细胞SGC-7901的生长抑制作用,Hoechst染色和Annexin V-FITC/碘化丙啶(PI)流式细胞术检测细胞凋亡情况,JC-1法检测线粒体膜电位变化,Western blotting法检测线粒体途径相关蛋白细胞色素C、Caspase-3和Caspase-9的表达。结果 MTT结果显示,24 h和48 h时不同组胃腺癌细胞SGC-7901吸光度(A)、生长抑制率比较,差异均有统计学意义(P<0.05);组间两两比较胃腺癌细胞SGC-7901 A值依次降低、生长抑制率依次升高(P<0.05)。荧光显微镜下观察发现,对照组细胞核显示为蓝荧光且较均匀,0.500μg/ml组处理12 h后细胞核呈紧密较亮荧光体,颜色较白。不同组胃腺癌细胞SGC-7901总相对死亡率比较,差异有统计学意义(P<0.05);组间两两比较胃腺癌细胞SGC-7901总相对死亡率依次升高(P<0.05)。JC-1染色结果显示,毒毛旋花子阿洛糖苷处理胃腺癌细胞SGC-7901 12 h后,0.125μg/ml组、0.250μg/ml组、0.500μg/ml组显示绿色荧光,随毒毛旋花子阿洛糖苷浓度的增加绿色荧光增强。Western blotting法显示,随着细胞凋亡的发生,细胞色素C从线粒体中释放到细胞质中,同时,Caspase-3和Caspase-9也被激活并发生剪切活化,活化片段表达增加。结论毒毛旋花子阿洛糖苷通过线粒体凋亡途径诱导体外培养的人胃腺癌细胞SGC-7901凋亡。Objectjve To explore the apoptosis - inducing effect of Strophalloside on gastric adenocarcinoma cells type SGC - 7901 and its mechanism. Methods The human gastric adenocarcinoma cells type SGC - 7901 were divided into control group,0. 125 μg/ ml group,0. 250 μg/ ml group,0. 500 μg/ ml group,and 1. 000 μg/ ml group. The growth inhibition effect of Strophalloside on gastric adenocarcinoma cells type SGC - 7901 was detected by MTT assay,and the apoptosis was measured by Annexin V - FITC/ PI flow cytometry and Hoechst staining. The changes of mitochondrial membrane potential were examined by JC - 1 kit. The expressions of mitochondrial pathway - related protein cytochrome C,Caspase - 3 and Caspase - 9 were detected by Western blotting analysis. Results According to MTT assay results,there were significant differences in value A and rate of growth inhibition of gastric adenocarcinoma cells type SGC - 7901 among different groups at 24 h and 48 h after treatment, respectively（P 〈 0. 05）;comparison of the above two indicators between groups showed that value A of gastric adenocarcinoma cells type SGC - 7901 decreased sequentially,and rate of growth inhibition increased sequentially（P 〈 0. 05）. By fluorescence microscope,the cell nucleus of control group showed homogeneous blue fluorescence,the cell nucleus of 0. 500 μg/ ml group showed compact bright fluorescence with white color. There was significant difference in total relative mortality of gastric adenocarcinoma cells type SGC - 7901 among different groups（ P 〈 0. 05）;comparison of the above indicator between groupsnbsp;showed that total relative mortality of gastric adenocarcinoma cells type SGC - 7901 increased sequentially（ P 〈 0. 05 ）. According to results of JC - 1 staining,gastric adenocarcinoma cells type SGC - 7901 showed green fluorescence 12 h after Strophalloside treatment,and fluorescence intensity increased as concentration of Strophalloside rised. According to Western blotting results,cytochrome C was released from mitochon

关 键 词：胃肿瘤 毒毛旋花子甙类 线粒体 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]