APO2.7在凋亡和坏死的HL-60细胞中的表达

Expression of APO2.7 in Apoptotic and Necrotic HL-60 Cells

目的 研究单克隆抗体ＡＰＯ２．７识别的一种线粒体膜蛋白质在凋亡和坏死的ＨＬ－６０细胞中的表达情况。方法 依托泊甙（ＶＰ１６）和热处理分别诱导ＨＬ－６０细胞产生凋亡和坏死，用荧光显微镜检查、ＤＮＡ片段化和ＰＩ摄取试验进行鉴定，利用流式细胞仪分析其ＡＰＯ２．７的动态表达。结果 凋亡细胞的ＡＰＯ２．７表达率在０、２、４、８、１６、２４ｈ分别为（２．５±０．５６）％、（４．４±０．６１）％、（６．５±０．７０）％、（６．２±０、１７）％、（２５．８±０．５６）％和（１５．７±０．２６）％。而坏死细胞却分别是（４．５±０．７２）％、（６２．８±０．２６）％、（８７．８±０．４１）％、（９３．５±０．９５）％、（９４．８±０．５３）％和（８９．３±０．８５）％。二者均在１６ｈ达到峰值。结论 提示ＡＰＯ２．７可能不是凋亡测定的特定标志而是在细胞死亡（凋亡和坏死）过程中发挥作用。

Objective To investigate the expression pattern of a kind of mitochondria mem- brane protein recognized by the APO2.7 monoclonal antibody in cells undergoing apoptosis and necrosis. Methods Apoptotic and necrotic HL - 60 cells were determined with fluorescene microscopy, DNA fragmentation and PI intake test, which were induced by VP16 and thermodynamics, respectively. Then, the cells were analyzed by flow cytometry to monitor the kinetics of APO2.7 expression Results At 0、2、4、8、16 and 24h, the positive population of apoptotic cells were (2.5 ± 0.56) %, (4.4 ± 0.61)%, (6.5 ± 0.70)%, (6.2 ± 0.l7)%, (25.8 ± 0.56) % and (15.7 ± 0.26) %, respectively. Meanwhile, necrotic cells were (4.5 ± 0.72)%, (62.8 ± 0.26) %, (87.8 ± 0.41)%, (93.5 ± 0.95)%, (94.8 ± 0.53) %, and (89.3 ± 0.85)%, respectively. Both apoptotic and necrotic cells reached the peak at 16h. Conclusion The data herein suggested that APO2.7 may not be a specific marker for apoptosis measurement but it plays a role in the cell death process (apoptosis and necrosis).