微波辐射诱导人神经母细胞瘤SH-SY5Y细胞凋亡的可能机制  

Mechanism of apoptosis induced by microwave radiation in human neuroblastoma SH-SY5Y cells

作　　者：王程[1,2] 李琴[3] 刘微[1] 闫岗林[2] 徐冶[1] 于洋[1] 刘师兵[1] 潘文干[1] 

机构地区：[1]吉林医药学院生物化学教研室,吉林吉林132013 [2]吉林大学分子酶工程教育部重点实验室,吉林长春130012 [3]中国环境科学研究院环境标准研究所,北京100012

出　　处：《吉林大学学报（医学版）》2011年第3期422-426,I0002,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅白求恩基金资助课题(200705408);吉林省教育厅科研基金资助课题(2009315);吉林医药学院科研基金资助课题(200903)

摘　　要：目的:观察微波辐射对人神经母细胞瘤SH-SY5Y细胞凋亡的影响,并探讨其机制。方法:采用10、30和50 mW.cm-2辐射强度的微波辐射SH-SY5Y细胞5 min,以假辐射组(0 mW.cm-2)为对照,光镜下观察细胞形态变化;荧光显微镜下观察DAPI染色细胞核的形态;CTAB法提取细胞基因组DNA,电泳观察DNAladder;台盼蓝拒染法检测细胞存活率;AnnexinV-FITC和PI双染色,流式细胞仪检测细胞凋亡率;MTT法检测细胞相对活性;蛋白质印迹检测细胞凋亡相关蛋白的表达。结果:微波辐射后SH-SY5Y细胞形态即刻改变,胞核、胞质结构欠清,细胞皱缩甚至脱壁;细胞核内的染色质出现不规则凝集,碎裂成2～5个微核;细胞DNA出现明显的梯状条带;细胞存活率随微波辐射强度增大逐渐降低,10 mW.cm-2辐射组与假辐射组比较,细胞存活率差异无统计学意义(P>0.05),但30、50 mW.cm-2辐射组细胞存活率明显低于假辐射组(P<0.05);微波辐射后6 h的细胞凋亡率随辐射强度的增加逐渐升高,各辐射组细胞凋亡率均明显高于假辐射组(P<0.05);微波辐射后24和48 h,细胞相对活力随辐射强度的增加明显降低,各辐射组的细胞相对活力均明显低于假辐射组(P<0.05)。微波辐射后,细胞凋亡相关蛋白Bcl-2、survivin表达水平逐渐下降,Bax、caspase-3和caspase-7表达水平逐渐升高,caspase-8和-9表达水平无明显变化。结论:微波辐射可诱导SH-SY5Y细胞凋亡,其作用机制可能与survivin、Bcl-2蛋白表达水平下调、Bax蛋白表达水平上调及caspase-3和caspase-7的作用有关,而与caspase-8和caspase-9介导的caspase-3活化这一作用无密切关系。Objective To observe the influence of microwave radiation in the apoptosis of human neuroblastoma SH-SY5Y cells and explore its mechanism.Methods Regarding 0 mW·cm-2 as sham radiation group,the SH-SY5Y cells were radiated for 5 minutes by 10,30 and 50 mW·cm-2 of microwave.The morphological changes of cells were observed under light microscope;the cell nucleus stained by DAPI were observed under fluorescence microscope;the cell genomic DNA was extracted by CTAB and electrophoresed to observe DNA ladder;the cell viability was detected by trypan blue exclusion assay;the apoptotic rate was detected by flow cytometry with annexin V-FITC/PI double staining;the relative activity of cells was detected by MTT assay;the expressions of apoptosis-related proteins were detected by Western blotting.Results After microwave radiation,the morphology of SH-SY5Y cells changed immediately,the nucleus and cytoplasm structure were less clear,and cell shrinkage or even off the wall were observed;chromatin in the nucleus appeared irregular condensation,and fragmentated into 2 to 5 micronuclei;cell DNA ladder was apparent.The survival rate of SH-SY5Y cells was reduced gradually with the increasing of microwave radiation intensity,there was no significant difference between 10 mW·cm-2 radiation group and sham radiation group（P0.05）;however,the cell survival rates in 30 and 50 mW·cm-2 radiation group were lower than that in sham radiation group（P0.05）.The apoptotic rate of SH-SY5Y cells was gradually increased with the microwave radiation intensity in 6 h after microwave radiation,and the apoptotic rates in each radiation group were higher than that in sham radiation group（P0.05）.Compared with sham radiation group,the cell relative activities of SH-SY5Y cells in each radiation group were markedly decreased 24 and 48 h after microwave radiation（P0.05）,and they were decreased with the increasing of microwave radiation intensity.After microwave radiation,the expressions of apoptosis related proteins Bcl-2 and survivin were

关 键 词：微波辐射 人神经母细胞瘤 SH-SY5Y细胞 细胞凋亡 

分 类 号：Q684[生物学—生物物理学]