泛素连接酶Cbl对TRAIL诱导Jurkat T细胞凋亡调节作用的探讨

Role of Cbl family of ubiquitin ligases in TRAIL-induced apoptosis of Jurkat T cells

目的:探讨泛素连接酶Cbl在TRAIL诱导Jurkat T细胞凋亡中的调节作用。方法:台盼蓝拒染法检测细胞增殖能力,流式细胞仪PI染色检测细胞凋亡,蛋白质印迹法检测蛋白的表达。结果:不同浓度的TRAIL作用于Jurkat T细胞24 h,TRAIL能以剂量依赖性的方式抑制JurkatT细胞增殖,并能诱导Jurkat T细胞凋亡。100 ng/mL的TRAIL作用24 h,有35.98%的细胞发生凋亡,而对照组仅有2.52%的细胞发生凋亡,P<0.01。在TRAIL诱导Jur-kat T细胞凋亡过程中伴随有p-Akt表达的一过性上调和泛素连接酶Cbl-b和c-Cbl表达的持续上调。结论:TRAIL上调Cbl蛋白的表达是抑制Jurkat T细胞中PI3K/Akt信号过度活化,进而诱导细胞凋亡的重要机制。

OBJECTIVE： To investigate the role of Cbl family of ubiquitin ligases in TRAIL-induced apoptosis of Jurkat T cells.METHODS： Cell proliferation was measured by trypan blue exclusion test and cell apoptosis was determined by flow cytometry using propidium iodide staining.Protein expression was assayed by Western blot.RESULTS： Treatment Jurkat T cells with different doses of TRAIL for 24 h,TRAIL could significantly inhibit Jurkat T cell proliferation in a classical dose-dependent manner.Jurkat T cells were exposed to 100 ng/mL TRAIL for 24 h,35.98% of cells underwent apoptosis,while only 2.52% cells underwent apoptosis in control group（P0.01）.During the process of TRAIL-induced apoptosis of Jurkat T cells,TRAIL transiently activated PI3K/Akt signaling and sustained upregulated Cbl-b and c-Cbl expression.CONCLUSION： TRAIL can increase the expression of Cbl proteins,which is an important mechanism to inhibit excessive activation of PI3K/Akt signaling,and thus to induce apoptosis of Jurkat T cells.