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GFI-1 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-α2b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。
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 syndrome-derived 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-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