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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。
Objective: To compare the sensitivity of fluorescent labeled Lactadherin and caspase-3 in the apoptosis of tongue squamous carcinoma cell line CAL-27 induced by As2O3, There is a better understanding of the different meanings of the two methods in testing results. Methods: The squamous cell carcinoma of the tongue, CAL-27, was treated with As2O3 at the final concentration for 0 h, 3 h, 6 h, 12 h respectively. The in situ activated caspase was labeled with fluorescently labeled caspase-3 inhibitor (FAM200-1) -3, fluorescence labeled Lactadherin647 detection of apoptosis, fluorescence inverted microscope analysis of its morphological changes, quantitative detection by flow cytometry and analysis of changes in its activity. RESULTS: After As2O3 induced cells for 3 h, 98.5% of cells were labeled with FAM200-1 and only 1.3% of cells were labeled with Lactadherin647. After 6 h and 12 h, the cells labeled with Lactadherin647 increased to 2.5% and 6.1%, respectively. Conclusion: As2O3-induced cells apoptosis after 3 h. FAM200-1 detects early apoptotic cells prior to lactadherin, and caspase-3 activation precedes PS turnover.