摘要
目的:研究发现Hela细胞经辐射后,其IER5的mRNA出现高表达,说明IER5基因与辐射之间存在的一定的量效关系[1-5]。为了进一步研究这一关系,开展了对IER5基因抑制及表达后,细胞经辐射反应影响的研究,其中包括对细胞凋亡现象进行的定量分析。方法:将Hela细胞与HepG2细胞接种在裸鼠皮下,进行饲养,一定时期后进行γ射线照射,取其脏器,制作成切片,经TUNEL技术染色后,在荧光显微镜下观察并拍摄细胞凋亡图片,对所拍摄的图片用MATLAB图像处理软件进行图像处理,并通过编程对大量的凋亡细胞进行自动计数。结果:对接种了Hela细胞与HepG2细胞的裸鼠经辐射后,其组织有细胞凋亡现象;对荧光显微镜下拍摄的细胞凋亡图片用MATLAB图像处理软件进行处理和编程,实现了对图片中的凋亡细胞的定量分析和自动计数。结论:接种了Hela细胞与HepG2细胞的裸鼠,经辐射后,其脑组织、肝组织切片经TUNEL染色后,在荧光显微镜下观察到细胞凋亡现象;用MATLAB数字图像处理软件可以对荧光显微镜下拍摄的细胞凋亡图片进行处理,并可以实现图片中凋亡细胞的定量分析。
Objective：The mRNA of IER5（Immidiate early response gene 5） in Hela cells could be up-regulated after radiation,this indicates that there is a biological relationship between IER5 and radiation.In order to study this relationship,RNAi technolo-gy and gene overexpression technology were employed to make the IER5 RNAi expression vector and overexpression vector to transfect to Hela cells,and further investigate the radiation effect on Hela cells.This study involved quantitative analysis to apop-tosis cells.Methods：Hela cells and HepG2 cells were inoculated to nude mice through hypodermic injection.They were fed and irradiated by γ radiation.The organs of the mice were later exteriorized and were cut into slices.After TUNEL technology,apoptosis were found under fluorescence microscope.In order to have a quantitative analysis on this apoptosis phenomenon,pic-tures of organ slices were taken using fluorescence microscope firstly,and then MATLAB graphic software were used to analyze the pictures.Finally,the numbers of apoptosis cells were calculated through programming.Results：To irradiate to the naked mice which were injected the Hela cells or HepG2 cells,we investigated the apoptosis cells.MATLAB graphic software was used to analyze the pictures.Finally,the number of apoptosis cells was calculated through programming.Conclusions：We removed the brain and liver tissues of naked mice which were injected the Hela or HepG2 cells and received the radiation,the tissues were stained with TUNEL method and evaluated the apoptotic circumstances under the fluorescence microscopy.MATLAB graphic software was used to analyze the pictures.Finally,the number of apoptosis cells was calculated through programming.
出处
《中国医学物理学杂志》
CSCD
2010年第5期2153-2156,共4页
Chinese Journal of Medical Physics
基金
北京市教委科技发展项目(No.KM200710025007)
国家自然科学基金项目(No:30400120)
北京市自然科学基金(No.7092013)
国家高技术研究发展计划"863"项目(No.2004AA221160)
首都医科大学基础-临床科研合作基金项目(No.2009JL06)与资助
