纳米SiO_2对人神经母细胞瘤SH-SY5Y细胞的凋亡诱导作用及其机制  

Induction effect of nano-SiO_2 on apoptosis of neuroblastoma SH-SY5Y cells and its mechanism

作　　者：杨艳艳[1] 金明华[1] 李艳博[2] 李阳[2] 王佳慧[1] 郑彤[1] 王思惠[1] 迟翔宇[1] 张宇佳[1] 孙志伟[1,2] 

机构地区：[1]吉林大学公共卫生学院卫生检验学教研室,吉林长春130021 [2]首都医科大学公共卫生学院卫生毒理与卫生化学学系,北京100069

出　　处：《吉林大学学报（医学版）》2015年第2期249-254,I0002,共7页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81172704);吉林大学大学生创新创业项目资助课题(20130943)

摘　　要：目的:探讨纳米二氧化硅(SiO2)对人神经母细胞瘤SH-SY5Y细胞凋亡和周期的影响,阐明其作用机制。方法:选取不同浓度粒径为90nm的SiO2颗粒作用于SH-SY5Y细胞,分别为0、3.125、6.250、12.500、25.000、50.000mg·L-1 SiO2组,其中0mg·L-1组为对照组。MTT法检测各组SH-SY5Y细胞存活率;采用AO/EB染色和AnnexinⅤ-FITC/PI法检测各组细胞凋亡率;流式细胞术(FCM)检测细胞周期百分比和细胞中活性氧(ROS)水平。结果:MTT检测,6.250、12.500、25.000和50.000mg·L-1 SiO2组细胞存活率低于对照组(P<0.05),12.500、25.000和50.000mg·L-1 SiO2组细胞存活率低于3.125mg·L-1 SiO2组(P<0.05)。AO/EB染色和Annexin V-FITC/PI法检测,6.250、12.500、25.000和50.000 mg·L-1 SiO2组细胞凋亡率高于对照组(P<0.05),12.500、25.000和50.000 mg·L-1 SiO2组细胞凋亡率高于3.125mg·L-1 SiO2组(P<0.05)。FCM检测,3.125、6.250、12.500、25.000和50.000mg·L-1 SiO2组细胞中ROS水平高于对照组(P<0.05)。25.000和50.000 mg·L-1 SiO2组G0/G1期细胞百分比低于3.125mg·L-1SiO2组和对照组(P<0.05),50.000 mg·L-1 SiO2组G2/M期细胞百分比高于对照组(P<0.05)。结论:纳米SiO2可降低SH-SY5Y细胞的活力,诱导细胞凋亡,增加细胞中ROS水平,可干扰细胞周期进程,对SH-SY5Y细胞的增殖具有抑制作用。Objective To observe the effects of nano-SiO2 on apoptosis and cell cycle of human neuroblastoma SHSY5 Y cells,and to explore the mechanism. Methods The SH-SY5 Y cells were treated with different concentrations of 90 nm SiO2 and divided into 0,3.125,6.250,12.500,25.000,and 50.000 mg·L-1 SiO2 groups.The survival rates of the cells in various groups were assessed by MTT method;the apoptotic rates of SHSY5 Ycells were determined by AO/EB staining and Annexin Ⅴ-FITC/P method;the percentage of cell cycle and the level of intracellular reactive oxygen species（ROS） were tested by flow cytometry（FCM） method.Results The MTT results showed the survival rates of the cells in 6.250,12.500,25.000 and 50.000mg·L-1SiO2 groups were significantly lower than that in control group（P〈0.05）;the survival rates of the cells in 12.500,25.000,and 50.000mg·L-1 SiO2 groups were significantly lower than that in 3.125mg·L-1 SiO2group（P〈0.05）;the AO/EB staining and Annexin Ⅴ-FITC/PI results showed the apoptotic rates of the cells in 6.250,12.500,25.000,and 50.000 mg·L-1 SiO2 groups were higher than that in control group（P〈0.05）;the apoptotic rates of the cells in 12.500,25.000,and 50.000 mg· L-1 SiO2 groups were higher than that in3.125mg·L-1 group（P〈0.05）.The FCM results showed the levels of ROS of the cells in 3.125,6.250,12.500,25.000,and 50.000mg·L-1 SiO2 groups were significantly higher than that in control group（P〈0.05）;compared with control group,the percentages of the cells at G0/G1 phase in 25.000 and 50.000 mg·L-1 SiO2 groups were increased（P〈0.05）;the percentage of the cells at G2/M phase in 50.000mg·L-1 SiO2 group was increasd（P〈0.05）.Conclusion Nano-SiO2 can decrease the activity of SH-SY5 Ycells and increase the apoptosis and the level of ROS,and nano-SiO2 can interfere the cell cycle and inhibit the proliferation of SH-SY5 Ycells.

关 键 词：纳米二氧化硅 SH—SY5Y细胞 细胞周期 细胞凋亡 

分 类 号：R730.264[医药卫生—肿瘤]