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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 investigate the effects of IFN-α on proliferation and apoptosis of human cutaneous T-cell lymphoma (CTCL) cells and to explore the mechanism of IFN-α in CTCL. Methods Hut78 cells were treated with IFN-α at 3000, 5000, 10000 and 20000U / mL for 24, 48, and 72 h, respectively. The survival rate of Hut78 cells was detected by MTS assay. Flow cytometry was used to detect the effect of IFN- After Hut78 cell apoptosis. The expression of BCL11B mRNA was detected by Real-time PCR at different time points and the expression of BCL11B protein was detected by Western blot at different time points. After IFN-α was detected by immunofluorescence at different time points, The expression and distribution of BCL11B protein in Hut78 cells. Results IFN-α significantly inhibited the proliferation of Hut78 cells in a dose-dependent and time-dependent manner. The inhibitory effect peaked at 48h. IFN-α also significantly induced apoptosis in Hut78 cells, and its effect was also concentration-dependent. The expression of mR-NA of BCL11B was significantly decreased after treated with 10000 U / mL IFN-α for 3, 6, 9 and 12 h, and the mRNA expression of BCL11B decreased to the lowest point at 6 h, After 12 and 24 h, the protein of BCL11B also decreased significantly, and the protein expression of BCL11B decreased to the lowest at 12h. CONCLUSION: IFN-α plays a therapeutic role in CTCL by inhibiting cell proliferation and promoting cell apoptosis. It can also promote the apoptosis of CTCL by decreasing the expression of BCL11B and enhance the sensitivity of CTCL cells to conventional chemotherapy.