成像流式细胞术在细胞自噬及血液学研究中的应用
首发时间:2014-12-09
摘要:本文初步探索了成像流式细胞仪在细胞自噬及血液学研究中的应用。细胞自噬水平通常用LC3聚集的程度来表示,荧光染色后,成像流式仪内置的Spot count模块可对细胞内聚集的荧光斑点数进行统计,得出一个细胞群体的整体的自噬水平。本文以急淋白血病细胞697为对象,饥饿HBSS以及Baf A1处理细胞,成像流式细胞仪检测细胞自噬水平,Spot count模块统计细胞内的自噬斑点数,结果显示饥饿可显著激活细胞自噬水平,斑点数从对照组的19.8±1.8 到饥饿组的37.4±3.9,(P<0.05)。Rapamycin及HBSS处理细胞,成像流式细胞仪检测白血病融合蛋白E2A/Pbx1与自噬蛋白LC3的共定位程度,Co-localization模块分析结果显示共定位百分比在对照,HBSS和Rapamycin组分别为17.5±0.8,23.3±1.2,28.1±2.9,(P<0.05),药物Rapamycin和饥饿HBSS可提高细胞蛋白的共定位程度,白血病融合蛋白可能通过自噬途径降解。
关键词: 细胞生物学 成像流式细胞术 细胞自噬 蛋白共定位 血液学
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Application of image flow cytometry in cell autophagy and hematology study
Abstract:This study explored the Application of image flow cytometry in cell autophagy and hematology study. Autophagy level is usually measured by autophagy plot of LC3 spot in cells. The LC3 spot count could be statisticaled by the spot count wizard in image flow cytometry and the cell autophagy level was analyzed. Acute lymphoblastic leukemia 697 cells were used in this study. Cells were treated with starvation of HBSS, Bafilomycin A1 or combined to measure the LC3 spot conut. It showed that the spot increased from 19.8±1.8 in ctrl to 37.4±3.9 in starvation(P<0.05),indicated that autophagy was activated significantly by starvation.The co-localization of leukemia oncoprotein E2A/Pbx1 with autophagy marker LC3 showed that the co-localization percentage in ctrl, rapamycin and HBSS were 17.5±0.8,23.3±1.2,28.1±2.9,respectively,(P<0.05),indicated that the leukemia oncoprotein was degraded might thorough autophagy mechanism.?????
Keywords: Cell biology image stream flow cytometry cell autophagy protein co-localizationl hematology
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成像流式细胞术在细胞自噬及血液学研究中的应用
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