岩藻黄素对脑胶质瘤细胞U87增殖及自噬的影响  

Effect of fucoxanthin on proliferation and autophagy of human glioma U87 cells

作　　者：刘钰罡 孙移坤 戴宜武[1] 

机构地区：[1]南方医科大学附属北京军区总医院临床学院附属八一脑科医院,北京100700 [2]解放军医学院附属北京军区总医院临床学院附属八一脑科医院

出　　处：《山东医药》2016年第15期4-7,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81271391)

摘　　要：目的观察岩藻黄素对脑胶质瘤细胞U87增殖及自噬的影响。方法未经处理的U87细胞作为对照组,不同浓度岩藻黄素处理的U87细胞作为实验组,比较各组细胞生存率、细胞凋亡率及自噬体、自噬溶酶体数目和分布,另比较细胞亚显微结构、微管相关蛋白1轻链3Ⅱ型/微管相关蛋白1轻链3Ⅰ型(LC3Ⅱ/LC3Ⅰ)、酵母自噬基因6同系物(Beclin1)。结果 12.5、25、50、75μmol/L岩藻黄素处理组细胞生存率分别为93.19%±5.88%、73.05%±2.33%、49.95%±1.59%、25.95%±1.40%,对照组为100.00%±1.35%,25、50、75μmol/L岩藻黄素处理组分别与对照组比较,P均<0.05。25、50μmol/L岩藻黄素处理组细胞凋亡率分别为12.00%±0.56%、31.97%±1.83%,对照组为2.97%±0.21%,各岩藻黄素处理组分别与对照组比较,P均<0.05。25、50μmol/L岩藻黄素处理组细胞LC3-GFP融合蛋白转移至自噬体膜并呈斑点状分布于细胞质内,且斑点数目随药物浓度的升高而增加;对照组LC3-GFP融合蛋白弥散分布在胞质中。镜下可见对照组正常的细胞器结构;50μmol/L岩藻黄素处理组可见大量的自噬小泡、自噬体及自噬溶酶体,其中可见尚未完全降解线粒体结构。25、50μmol/L岩藻黄素处理组LC3Ⅱ/LC3Ⅰ分别为0.86±0.05、0.89±0.04,对照组为0.65±0.01,各岩藻黄素处理组分别与对照组比较,P均<0.05;25、50μmol/L岩藻黄素处理组Beclin1相对表达量分别为0.39±0.02、0.59±0.04,对照组为0.23±0.01,各岩藻黄素处理组分别与对照组比较,P均<0.05。结论岩藻黄素可有效抑制U87细胞增殖,并诱导自噬的发生。Objective To observe the effect of fucoxanthin on the proliferation and autophagy of glioma U87 cells. Methods The untreated U87 cells were taken as the control group, and U87 cells treated with different concentrations of fucoxanthin were taken as the experimental group. The viability, the apoptosis rate, the number and distribution of autopha-gosome and autolysosome were compared between different groups. Meanwhile, the cell ultrastructure, microtubule-associ-ated protein 1 light chain 3Ⅱ/microtubule-associated protein 1 light chain 3Ⅰ（ LC3Ⅱ/LC3Ⅰ） , and Beclin-1 were com-pared between the two groups. Results The cell survival rates of the experiment groups treated with 12. 5, 25, 50 and 75μmol/L fucoxanthin were 93. 19% ± 5. 88%, 73. 05% ± 2. 33%, 49. 95% ± 1. 59% and 25. 95% ± 1. 40%, respective-ly. The viability of the control group was 100. 00% ± 1. 35%. Significant difference was found between the experimental groups treated with 25, 50, 75μmol/L fucoxanthin and the control group （all P〈0. 05）. The apoptosis rates of the exper-imental groups treated with 25 and 50 μmol/L fucoxanthin were respectively 12. 00% ± 0. 56% and 31. 97% ± 1. 83%, and it was 2. 97% ± 0. 21% in the control group （all P〈0. 05）. Fucoxanthin induced autophagy in dose-dependent man-ner, which was assayed by transient transfection of LC3-GFP. Compared with the control group, there were more lysosome and autophagolysosomes, indicated by LC3-GFP, in U87 cells treated with fucoxanthin （25 and 50μmol/L）. Fucoxanthin-treated cells displayed many lysosome and autophagolysosomes which were observed by transmission electron microscopy. In the experiment groups treated with fucoxanthin （ 25 and 50 μmol/L ） , the expression of LC3Ⅱ/LC3Ⅰand Beclin1 was 0. 86 ± 0. 05, 0. 89 ± 0. 04 and 0. 39 ± 0. 02, 0. 59 ± 0. 04. In the control group, the expression of LC3Ⅱ/LC3Ⅰand Bec-lin1 was 0. 65 ± 0. 01 and 0. 23 ± 0. 01. Compared with the control group, autophagic-related protein expression of LC3Ⅱ/

关 键 词：岩藻黄素 胶质瘤 微管相关蛋白1轻链3 Ⅱ型 微管相关蛋白1轻链3 Ⅰ型 酵母自噬基因6同系物 

分 类 号：R739.4[医药卫生—肿瘤]