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人重组TFAR19蛋白对白血病细胞株HL |
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张颖妹 徐秀珍 刘红涛 宋泉声 马大龙
摘 要：目的：对一种新发现的凋亡相关基因TFAR19进行蛋白质水平的促凋亡效应研究，并对其作用机理进行初步的探讨。 方法：利用离子交换层析对大肠杆菌表达的人重组TFAR19蛋白进行纯化，将其加入到培养的白血病细胞株HL-60，通过DNA片段化、PI及Annexin V标记进行流式细胞仪分析，观察TFAR19蛋白的促凋亡效应。利用FITC标记TFAR19蛋白，分析其与细胞的结合及定位。利用Caspase-3抑制剂DEVD-fmk研究TFAR19的凋亡信号转导途径。结果：高纯度的重组TFAR19蛋白剂量依赖性地促进撤除血清的HL-60细胞的凋亡，最高可使82%细胞凋亡，对照为30%。荧光标记的TFAR19蛋白能与HL-60细胞结合，主要定位于细胞核。DEVE-fmk可部分抑制TFAR19蛋白的促凋亡效应。结论：TFAR19蛋白能够直接进入HL-60细胞，发挥其促进细胞凋亡的效应。这一效应部分地与Caspase-3的凋亡执行效应有关。
关键词：TFAR19 凋亡 HL-60细胞 Caspase-3
分类号：R371
The apoptosis-accelerating effect of human recombinant TFAR19 protein on leukemia HL-60 cells
ZHANG Ying-Mei
Laboratory of Medical Immunology, Beijing Medical University, Beijing 100083
XU Xiu-Zhen
Laboratory of Medical Immunology, Beijing Medical University, Beijing 100083
LIU Hong-Tao
Laboratory of Medical Immunology, Beijing Medical University, Beijing 100083
Abstract：Objective: To study the apoptosis-accelerating effect of a novel human protein named TFAR19 (TF-1 cells apoptosis related protein 19) whose encoding cDNA has been cloned in the laboratory, and try to find out the mechanism of its effect. Methods： By using ion exchange chromatography, the human recombinant TFAR19 protein was isolated. The purified protein was co-cultured with HL-60 cells and the apoptotic cells were analyzed by DNA fragmentation, PI and Annexin V-labeled FACS assay. The FITC labeled TFAR19 protein w used as probe to observe its localization within the HL-60 cells. For the signal transduction study, a caspase-3 inhibitor DEVD-fmk was used to block the TFAR19 effect. Results： The TFAR19 protein dose-dependently increases the apoptotic HL-60 cells which have been previously withdrawn serum from culture, up to 82% cells are apoptotic cells comparing with control 30% cells. FITC labeled TFAR19 protein binds with HL-60 cells and mainly localized within the nucleus. DEVD-fmk partly inhibits apoptosis-acceleratin[1] [2] [3] 下一页
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