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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ＡｎｎｅｋｉｎＶ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ＴＦＡＮ１９蛋白能与ＨＬ－６０细胞结合，主要定位于细胞核。ＤＥＶＫ－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。
OBJECTIVE: To investigate the pro-apoptotic effect of a newly discovered apoptosis-related gene, TFAR19, at the protein level and to discuss its mechanism of action. Methods: The recombinant human TFAR19 protein expressed in E. coli was purified by ion exchange chromatography and added to the cultured leukemia cell line HL-60. Flow cytometric analysis was performed by DNA fragmentation, PI, and Annekin V labeling to observe TFAR19. The pro-apoptotic effects of proteins. FITC-tagged TFAR19 protein was used to analyze its binding and localization to cells. The apoptosis signal transduction pathway of TFAR19 was studied using Caspase-3 inhibitor DEVD-fmk. RESULTS: High-purity recombinant TFAR19 protein promoted the apoptosis of HL-60 cells in a dose-dependent manner. The highest level of apoptosis was in 82% of the cells, and the control was 30%. The fluorescently labeled TFAN19 protein binds to HL-60 cells and is mainly located in the nucleus. DEVK-fmk partially inhibits the pro-apoptotic effects of TFAR19 protein. Conclusion: TFAR19 protein can directly enter HL-60 cells and exert its effect of promoting apoptosis. This effect is partly related to the apoptotic effect of Caspase-3.