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目的病毒来源的凋亡蛋白Apoptin诱导不同类型肿瘤细胞的凋亡而对正常细胞无损伤。通常,机体系统治疗往往因其对快速分裂细胞组织尤其是对免疫细胞体系的毒性而受到制约。因此,本文旨在研究小鼠转基因Apoptin是否在正常淋巴细胞的发育、激活和增殖过程中对其具有干扰性。方法建立H-2K~b启动子表达调控的Apoptin转基因小鼠模型。以Northern和Western印迹法分析Apoptin在转基因小鼠不同组织中的表达。以FACS方法分析Apoptin的表达对B、T淋巴细胞的细胞效应。并以脂多糖(LPS)刺激Apoptin转基因细胞和对照细胞,比较生长曲线。结果 Apoptin主要表达于淋巴组织。重要的是,Apoptin不影响转基因小鼠的B、辅助T或细胞毒T细胞生长数量,提示诸细胞类型对Apoptin凋亡效应的非敏感性。蛋白酶体抑制实验表明,在这些正常细胞中,Apoptin呈短半衰期形式,该现象部分解释了Apoptin的肿瘤特异性机制。此外,LPS对B细胞的激活或IL-2和Con A对T细胞的刺激,不导致转基因淋巴细胞的生长劣势,亦不导致细胞死亡效应的增加。结论 Apoptin转基因小鼠的淋巴细胞在发育和增殖过程中对Apoptin的表达具有耐受性,为发展Apoptin的体系统肿瘤治疗消除了首要障碍。
Purpose Apoptosis protein Apoptin induced different types of tumor cells apoptosis without damage to normal cells. In general, systemic therapy is often limited by its toxicity to rapidly dividing cell tissues, especially to immune cell systems. Therefore, this article aims to investigate whether mouse transgenic Apoptin is interfering with normal lymphocyte development, activation and proliferation. Methods Apoptin transgenic mice model with H-2K ~ b promoter expression was established. The expression of Apoptin in different tissues of transgenic mice was analyzed by Northern and Western blotting. The cellular effects of Apoptin expression on B and T lymphocytes were analyzed by FACS. Apoptin transgenic cells and control cells were stimulated with lipopolysaccharide (LPS), and the growth curves were compared. Results Apoptin was mainly expressed in lymphoid tissues. Importantly, Apoptin does not affect the amount of B, helper T or cytotoxic T cell growth in transgenic mice, suggesting the non-sensitivity of cell types to the apoptotic effect of Apoptin. Proteasome inhibition experiments show that Apoptin exhibits a short half-life form in these normal cells, a phenomenon that partially explains the tumor-specific mechanism of Apoptin. In addition, activation of B cells by LPS or stimulation of T cells by IL-2 and Con A do not result in the growth disadvantage of transgenic lymphocytes and do not lead to an increase in cell death effects. Conclusion Lymphocytes in Apoptin transgenic mice are resistant to the expression of Apoptin during development and proliferation, and eliminate the primary obstacle for the development of systemic tumor therapy of Apoptin.