【摘要】 目的:比较荧光标记的乳粘素(Lactadherin)和半胱氨酸天冬肽酶3(caspase-3)在检测由三氧化二砷(As2O3)诱导舌鳞癌细胞CAL-27凋亡中的敏感性,以期对两种方法在检测结果方面的不同意义有进一步的了解。方法:将配置好终浓度的As2O3分别作用舌鳞癌细胞CAL-27 0 h、3 h、6 h、12 h,用荧光标记的caspase-3抑制剂(FAM200-1)标记原位激活的caspase-3,荧光标记的Lactadherin647检测细胞凋亡,荧光倒置显微镜分析其形态学变化,流式细胞仪定量检测并分析其活性变化。结果:As2O3诱导细胞3 h后,98.5%细胞被FAM200-1标记,只有1.3%细胞被Lactadherin647标记。6 h、12 h后,被Lactadherin647标记的细胞分别增至2.5%、6.1%。结论:As2O3作用细胞3 h后细胞发生凋亡。FAM200-1先于乳粘素检测出早期凋亡的细胞,caspase-3的激活早于PS翻转。更多还原

【Abstract】 Objective: To investigate the sensitivity of fluorescent lactadherin and cystatin radon peptidase 3(caspase-3) in detecting the apoptosis of tongue squamous carcinoma CAL-27 cells induced by arsenic trioxide in order to establish a more effective evaluation measure of apoptosis. Methods: As2O3was incubated with tongue squamous carcinoma CAL-27 cells for 0 h, 3 h, 6 h, 12 h.The activated caspase-3 was tested with the caspase-3 inhibitor(FAM200-1) in situ and the apoptotic cells were detected with fluorescent lactadherin. Fluorescence microscope was used to observe the morphological changes and flow cytometry to test in quantity as well as to analyse the activity of cells. Results: After incubation with As2O3for three hours, the percentage of cells marked with FAM200-1was 98.5% while only 1.3 % were positive for Lactadherin647. The cells labled by Lactadherin647 were increased to 2.5 % and 6.1 % respectively after 6 or 12 hours incubation. Conclusion: The apoptosis of CAL-27 cells began at 3 hours of inducement by arsenic trioxide. FAM200-1had the priority over lactadherin in detecting the early apoptotic cells, and PS externalization was delayed compared with early caspase-3activation.更多还原