摘要
目的探究绿原酸对胃癌SGC-7901细胞增殖及凋亡诱导因子(Apoptosis inducing factor,AIF)表达的影响。方法体外培养胃癌SGC-7901细胞24 h后,分别加入不同浓度的绿原酸处理细胞12、24、48、72 h,用四甲基偶氮唑蓝(MTT)法测定绿原酸对SGC-7901细胞抑制率,选取抑制率变化明显的3个浓度进行后续实验。将实验分为4组:对照组、低浓度组(19.375μg/m L)、中浓度组(38.75μg/m L)、高浓度组(77.5μg/m L),倒置荧光显微镜观察线粒体膜电位变化,qRTPCR及Western blot法检测不同浓度绿原酸作用于细胞后AIF的表达水平。结果绿原酸浓度越高,对SGC-7901细胞的生长抑制率越高,绿原酸浓度为19.375、38.75、77.5μg/m L时,抑制率变化最明显(P<0.01);绿原酸作用时间越长,对SGC-7901细胞的生长抑制率越高;与对照组相比,用药组随着用药浓度的增加,红色荧光逐渐降低,绿色荧光逐渐增强;与对照组相比,用药组AIF基因的表达显著增加(P<0.01),细胞质中AIF蛋白的表达显著增加(P<0.01);线粒体中AIF蛋白的表达显著降低(P<0.01)。结论绿原酸能抑制胃癌SGC-7901细胞的增殖,并通过降低线粒体膜电位和促进AIF从线粒体转移至细胞质,可能诱导细胞凋亡。
Objective To studythe effects of chlorogenic acid on the proliferation of gastric cancer SGC-7901 cells and the expression of apoptosis inducing factor（ AIF）. Methods After in vitro culture of gastric cancer SGC-7901 cell for 24 h,the cells were treated with different concentrations of chlorogenic acid for 12,24,48,72 h,respectively. The SGC-7901 cell inhibition was tested by thiazolyl blueterazolium bromide（ MTT） assay. Select three concentrations that had obvious changes in inhibition rate were selected for the subsequent experiments. The experiment was divided into four groups： control group,chlorogenic acid low concentration（ 19. 375 μg/m L）group,middle concentration（ 38. 75 μg/m L） group and high concentration（ 77. 5 μg/m L） group. The mitochondrial membrane potential was observed by inverted fluorescence microscope. qRT-PCR and western blot methods were used to detect the expression of AIF in cells. Results Compared with the control group,the high the concentration of chlorogenic acid induced the higher the growth inhibition rate of SGC-7901 cells. When the chlorogenic acid concentration was 19. 375,38. 75 and 77. 5 μg/m L,the inhibition rate was changed most significantly（ P〈0. 01）. The longer chlorogenic acid acted,the higher growth inhibition rate of SGC-7901 cells was shown. Compared with the control group,the red fluorescence in the treatment group was gradually decreased and the green fluorescence in the treatment group was gradually increased with the increased drug concentration. Compared with the control group,the expression of AIF gene in the treatment group was increased significantly（ P〈0. 01）,the expression of AIF protein in the cytoplasm was increased significantly（ P〈0. 01）,and the expression of AIF protein in mitochondria was decreased significantly（ P〈0. 01）. Conclusion Chlorogenic acid could inhibit the proliferation of gastric cancer SGC-7901 cells,and may induce cell apoptosis by reducing the mitochondrial membrane potential and promoting AIF protein transfer from mitochondrial to cytoplasm.
作者
白建伟
王峰
潘鸿
赵兰
粟立羽
李岩
Bai Jianwei;Wang Feng;Pan Hong;Zhao Lan;Su Liyu;Li Yan(Department of Occupational Health and Environmental Hygiene,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Labor Hygiene and Environmental Hygiene,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Food and Nutritional,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Health Toxicology,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China)
出处
《遵义医学院学报》
2018年第2期150-155,共6页
Journal of Zunyi Medical University
基金
贵州省科学技术基金资助项目(NO:黔科合LH字[2015]7555)
遵义医学院硕士启动基金资助项目(NO:F-716)
关键词
绿原酸
胃癌
凋亡诱导因子
细胞凋亡
Chlorogenic acid
gastric cancer
apoptosis inducing factor
cell apoptosis
