重组免疫毒素hIL2-Luffin P1对Hut-78细胞增殖及凋亡的影响

Effects of recombinant immunotoxin hIL2-Luffin P1 on proliferation and apoptosis of hut-78 cells

目的观察并评价重组免疫毒素hIL2-Luffin P1对皮肤T细胞淋巴瘤细胞(Hut-78)增殖和凋亡的影响。方法IL-2受体α链(CD25)抗体通过免疫细胞化学技术检测Hut-78细胞表面CD25分子的表达。将不同浓度hIL2-Luffin P1分别处理Hut-78细胞,用MTT法检测细胞增殖的抑制率,以Annexin V/PI双标法流式细胞术检测Hut-78细胞的凋亡。结果CD25在Hut-78细胞表面表达阳性率为79.32%。经浓度为0.5、1、10、20、30和40μg/mL的hIL2-Luffin P1作用48h,其对Hut-78细胞增殖的抑制率分别为11.03±1.604、19.21±1.577、32.41±1.744、42.37±1.289、52.80±2.031、59.33±1.261。经浓度为1、10、30μg/mL的hIL2-Luffin P1作用48h,Hut-78细胞的凋亡率分别为8.23±1.29、15.37士0.98和27.33±1.06。结论hIL2-Luffin P1能够抑制Hut-78细胞的增殖并能诱导凋亡。这提示重组免疫毒素hIL2-Luffin P1对具有IL-2受体表达的皮肤T细胞淋巴瘤可能具有潜在治疗作用。

Objective To investigate and evaluate the effects of recombinant immunotoxin hIL2-Luffin P1 on proliferation and apoptosis of T lymphoma cell （Hut-78 cells）. Methods CD25 expression on cell surface of Hut-78 cells was detected by immunocytochemistry with the antibody against human IL2 receptor α chain （CD25）. Hut-78 cells were treated with hIL2-Luffin P1 at different concentrations. Cell proliferation and apoptosis of Hut-78 cells was assessed respectively by MTT assay and Annexin V/PI doubled-labeled flow cytometry. Results The percentage of CD25 positive cells in Hut-78 cells was 79.32%. Inhibitory rate of Hut-78 cells was respectively 11.03±1. 604,19.21±1. 577,32.41±1. 744,42.37±1. 289,52.80±2. 031,59.33±1. 261, after the treatment with hIL2-Luffin P1 at concentration of 0. 5,1,10,20,30,40 μg/mL for 48h. Apoptotic rate of Hut-78 cells was respectively 8.23±1.29,15.37±0.98,27.33±1.06 after the treatment with hIL2-Luffin P1 at concentration of 1, 10 and 30 μg/ml for 48h. Conclusion hIL2-Luffin P1 could inhibit cell proliferation and induce cell apoptosis in Hut-78 cells. This suggests that hIL2- Luffin P1 may have potential therapeutic effect on cutaneous T cell lymphoma with expression of IL2 receptor.