IFN-α对人皮肤淋巴瘤细胞系Hut78的影响  

The Effects of IFN-α on Human Cutaneous T-cell Lymphoma Cell Line Hut78

作　　者：谷晓广[1] 汪旸[1] 张高磊[1] 涂平[1] 

机构地区：[1]北京大学第一医院皮肤性病科,北京100034

出　　处：《中国皮肤性病学杂志》2012年第10期871-874,879,共5页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金(编号:81072233)

摘　　要：目的研究IFN-α对人皮肤T细胞淋巴瘤(CTCL)细胞的增殖抑制和凋亡诱导作用,探讨IFN-α治疗CTCL的机制。方法分别用3000,5000,10000,20000U/mL的IFN-α作用Hut78细胞24,48,72h后,利用MTS法检测Hut78细胞的生存率;并用流式细胞术检测不同浓度IFN-α作用细胞24h后,Hut78细胞的凋亡情况。运用Real-time PCR检测不同时间点IFN-α作用后,BCL11B mRNA的表达情况,Western blot检测不同时间点IFN-α作用后BCL11B蛋白的表达情况,免疫荧光检测IFN-α不同时间点作用后,BCL11B蛋白在Hut78细胞表达程度和分布情况。结果 IFN-α能明显抑制Hut78细胞的增殖,且这种作用呈剂量依赖性和时间依赖性。其抑制效应在48h达到峰值。IFN-α还能显著诱导Hut78细胞的凋亡,其作用亦呈浓度依赖性。10000U/mL IFN-α作用细胞3,6,9,12h后,BCL11B mR-NA表达均显著降低,其中在6h时间点BCL11B的mRNA表达下降到最低点,10000U/mL IFN-α作用细胞6,12,24h后BCL11B的蛋白也明显降低,在12h时间点,BCL11B的蛋白表达量下降至最低。结论 IFN-α通过抑制细胞增殖和促进细胞凋亡对CTCL起到治疗作用,并可通过降低BCL11B的表达,促进细胞凋亡,并提高CTCL细胞对常规化疗的敏感性。Objective To study the effects of IFN_α on the proliferation and apoptosis of human cutaneous T cell lymphoma modeled cell line Hut-78, and to explore the molecular mechanism underlying it. Methods Hut78 cells were incubated with 3 000U ,5 000U, 10 000U ,20 000U/ml IFN_α for 24, 48, 72 hours. Cell viability assays were performed to measure MTS-based cell viability at each time points. Cell apoptosis after IFN_α incubation was quantified by flow cytometry analysis. Quantification of BCL11B expression on both mRNA and protein level was measured with quantitative realtime RT-PCR and western blot analysis before and after IFN_α treatment. Immunofluorescence analysis was used to observe the BCL11B expression on Hut-78 cell before and after IFN_α treatment. Results IFN_α significantly inhibited the growth of Hut78 cells. The effect was concentration dependent and duration dependent with the lowest growth rate at the time point of 48 hours. Meanwhile, IFN_α induced cell apoptosis, also in a concentration dependent way, with ceils treated with the highest concentration demonstrated the highest apoptosis rate. Furthermore, 10 000U/ml IFN_α incubated cells 3,6,9,12h time point, the BCL11B mRNA was significantly reduced. Specially, the BCL11B mRNA expression was at lowest point at 6h time point. 10 000U/ml IFN_α incubated cells 6,12,24h time point, the BCL11B protein also significantly reduced. The BCL11B protein expression decreased to a minimum at 12h time point. Conclusion IFN_α may exert its therapeutic effects on CTCL by directly inhibiting cell proliferation and inducing cell apoptosis. IFN_α may induce cell apoptosis and resensitize CTCL cells to chemotherapy by suppressing the expression of BCL11B gene.

关 键 词：IFN-Α 皮肤T细胞淋巴瘤 蕈样肉芽肿 SEZARY综合征 Hut78 BCL11B 

分 类 号：R733.4[医药卫生—肿瘤]