| 杨业鹏,朱应葆,陈冠英,周梅,沈嵚剑,沈磊.辐射诱发细胞凋亡4种定量检测方法的比较[J].中华放射医学与防护杂志,2004,24(6):497-499 |
| 辐射诱发细胞凋亡4种定量检测方法的比较 |
| Comparative study on 4 quantitative detection methods of apoptosis induced by radiation |
| 投稿时间：2004-07-15 |
| DOI： |
| 中文关键词: γ射线 肿瘤细胞 凋亡 坏死 检测方法比较 |
| 英文关键词:rays Tumor cells Apoptosis Necrosis Comparison between detecting methods |
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| 中文摘要: |
| 目的 通过检测结果的比较和原理分析,提示4种凋亡检测方法鉴别凋亡和坏死的能力及优缺点。方法 采用PI染色流式细胞测量(P-F)法、TUNEL标记流式细胞测量(T-F)法、AnnexinVFITC/PI活染流式细胞测量(A-F)法和Hoechst/PI活染荧光显微镜观察(H-O)法对γ射线诱发的人乳腺癌MCF7细胞凋亡和坏死进行检测,以羟基喜树碱和叠氮钠分别为诱发凋亡和坏死的阳性对照物。结果 4种方法对γ射线、羟基喜树碱诱发的细胞凋亡显示良好的时间剂量依赖性响应。P-F法所得凋亡率较低,曲线斜率最小;TF法反之;A-F法和HO法可获得凋亡和坏死两套数据,且两种方法的数值相近。A-F法和H-O法可识别叠氮钠所致的细胞坏死;而P-F法和T-F法不能反映其致坏死为主的真实情况。结论 P-F法和TF法不能区别凋亡和坏死,A-F法和HO法可以区别。PF法较TF法灵敏度低但简便经济。AF法快速可靠,H-O法经济实用并可同时观察细胞形态学变化。 |
| 英文摘要: |
| Objective To reveal the capability of 4 apoptosisdetecting Methods to discriminate between apoptosis and necrosis and show their respective advantages and shortcomings through comparison of detected Results and analysis of detection mechanism. Methods Four Methods,PI stainingflow cytometric detection (P-F method),TUNEL labelingflow cytometric detection (T-F method),annexin VFITC/PI vital stainingflow cytometric detection (A-F method) and Hoechst/PI vital stainingfluorescence microscopic observation (H-O method),were used to determine apoptosis and necrosis in human breast cancer MCF7 cell line induced by γ-rays.Hydroxycamptothecine and sodium azide were used to induce positive controls of apoptosis and necrosis respectively. Results All 4 Methods showed good timedependent and dosedependent respondences to apoptosis induced by γ-rays and hydroxycamptothecine.Apoptotic cell ratios and curve slopes obtained from PF method were minimum and,on the contrary,those from TF method were maximum among these 4 Methods.With AF method and HO method,two sets of data,apoptosis and necrosis,could be gained respectively and the data gained from these two Methods were close to equal.A-F method and HO method could distinguish necrosis induced by sodium azide from apotosis while P-F method and T-F method presented false increase of apoptosis. Conclusions P-F method and T-F method can not discriminate between apoptosis and necrosis.P-F method is less sensitive but more simple,convenient and economical than T-F method.AF method and HO method can distinguish necrosis from apoptosis.AF method is more costly but more quick and reliable than H-O method.H-O method is economical,practical and morphological changes of cells and nucleus can be observed simultaneously with it. |
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