独立生长因子1在Sézary综合征患者外周血Sézary细胞中的表达

目的 检测独立生长因子1(GFI-1)在Sézary综合征患者和正常人外周血中的表达,为开发针对GFI-1基因靶点的治疗提供实验依据.方法 利用流式细胞术分离纯化7例Sézary综合征患者外周血中CD4+CD7-的Sézary细胞(SS细胞)作为实验组,以10例正常人外周血CD4+T细胞、Sézary综合征来源细胞系Hut78细胞和人急性T细胞白血病细胞系Jurkat细胞为对照组.应用qPCR检测各组细胞中GFI-1 mRNA的表达,蛋白质印迹法检测GFI-1蛋白表达.用干扰素α2b(IFN-α2b)诱导Hut78细胞凋亡后,采用MTS法测定细胞增殖情况,用qPCR检测GFI-1、细胞周期依赖性蛋白激酶抑制因子P21、肿瘤坏死因子相关的凋亡诱导配体(TRAIL)和Caspase-3 mRNA的表达情况,用流式细胞术检测细胞凋亡情况.结果 Sézary综合征患者外周血SS细胞GFI-1 mRNA表达水平高于Jurkat细胞和正常人外周血CD4+T细胞(P均＜0.05).SS细胞和Hut78细胞的GFI-1蛋白表达水平高于Jurkat细胞和正常人外周血CD4+T细胞(P均＜0.05).IFN-α2b能够抑制Hut78细胞增殖,且其抑制作用呈时间和浓度依赖性.IFN-α2b处理Hut78细胞12 h和24 h后GFI1 mRNA的表达水平呈时间依赖性降低,P21、TRAIL和Caspase-3 mRNA的表达水平呈时间依赖性增加(P＜0.05).IFN-α2b处理Hut78细胞12 h和24 h后细胞的凋亡水平增加(P＜0.05).结论 GFI-1基因在Sézary综合征患者外周血SS细胞中表达增加,IFN-a2b能抑制Hut78细胞GFI-1基因的表达,表明GFI-1基因可能在Sézary综合征患者SS细胞的肿瘤性增殖中发挥重要调控作用.

Expression of growth factor independence 1 in peripheral blood Sézary cells of patients with Sézary syndrome

Objective To detect the expression of growth factor independence 1 (GFI-1) in peripheral blood of patients with Sézary syndrome and normal persons,so as to provide a theoretical basis for developing GFI-1 gene target therapy.Methods CD4+CD7-Sézary cells (SS cells) were separated and purified from peripheral blood of 7 patients with Sézary syndrome by flow cytometry,CD4+ T cells from peripheral blood of 10 normal persons,Sézary syndromederived cell line Hut78 and human acute T cell leukemia cell line Jurkat as controls.The mRNA and protein expressions of GFI-1 were detected by qPCR and Western blotting,respectively.Then after interferon-p-α-2b (IFN-α2b) was used to induce Hut78 cell apoptosis,the cell proliferation was measured by MTS,the mRNA expression of GFI-1,cell cycle dependent protein kinase inhibitor P21,tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Caspase-3 was detected by qPCR,and the cell apoptosis was detected by flow cytometry.Results The expression of GFI-1 mRNA in the SS cells was significantly higher than that in the Jurkat and CD4+T cells (all P＜0.05).The expression of GFI-1 protein in the SS cells and Hut78 cells was significantly higher than that in the Jurkat and CD4+T cells (all P＜0.05).IFN-α2b significantly inhibited the proliferation of Hut78 cells,and the effect was concentration-dependent and time-dependent.The mRNA expression of GFI-1 in Hut78 cells was significantly decreased in a time-dependent manner at 12 h and 24 h treated with IFN-α2b,while the mRNA expressions of P21,TRAIL and Caspase-3 were significantly increased (P＜0.05).The apoptosis of Hut78 cells was significantly increased at 12 h and 24 h treated with IFN-α2b (P＜0.05).Conclusion The expression of GFI-1 gene in peripheral blood SS cells of patients with Sézary syndrome is increased and can be inhibited by IFN-cα2b,indicating that GFI-1 gene may play an important regulatory role in tumor proliferation of SS cells in patients with Sézary syndrome.