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目的研究IFN-α联合苦参碱对皮肤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联合苦参碱治疗CTCL的机制。方法分别用3 000,5 000,10 000,20 000U/m L IFN-α;0.2,0.4,0.8,1.6mg/m L苦参碱;以及10 000U/m L IFN-α联合0.8mg/m L苦参碱分别作用HH细胞24,48,72h后利用MTS法检测HH细胞的生存率。并用流式细胞术检测10 000U/m L IFN-α,0.8mg/m L苦参碱以及其联合作用细胞24h后,HH细胞的凋亡情况。运用Real-time PCR检测10 000U/m L IFN-α,0.8mg/m L苦参碱以及其联合作用HH细胞24h后凋亡相关基因Fas,Fas L,Caspase8,Caspase3,Bax以及Bcl-2 mRNA的表达情况。结果 IFN-α和苦参碱单独作用均能有效抑制HH细胞的增殖,并引起HH细胞的凋亡。IFN-α联合苦参碱比单药对HH细胞的增殖抑制和凋亡诱导作用更明显,IFN-α联合苦参碱能使Fas,Caspase8,Caspase3以及Bax表达明显增加,能明显抑制Bcl-2的表达,但对Fas L的表达没有影响。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。与单独使用IFN-α相比,IFN-α联合苦参碱能明显提高CTCL的治疗效果,减少IFN-α的使用量。
OBJECTIVE To investigate the effects of IFN-α and matrine on the proliferation and apoptosis induction of cutaneous T-cell lymphoma (CTCL) cells and to explore the mechanism of IFN-α and matrine in the treatment of CTCL. Methods Matrine with 3 000, 5 000, 10 000 and 20 000 U / mL IFN-α, 0.2, 0.4, 0.8 and 1.6 mg / m L respectively and 10 000 U / L matrine, respectively, HH cells 24,48,72 hours after the use of MTS assay HH cell survival rate. Flow cytometry was used to detect the apoptosis of HH cells after 10 000 U / mL IFN-α, 0.8 mg / mL matrine and their combined action for 24 h. The apoptosis-related genes Fas, Fas L, Caspase8, Caspase3, Bax and Bcl-2 mRNA were detected by Real-time PCR after treated with 10 000 U / mL IFN-α and 0.8 mg / The expression of the situation. Results Both IFN-α and matrine alone could effectively inhibit the proliferation of HH cells and induce the apoptosis of HH cells. The combination of IFN-α and matrine significantly inhibited the proliferation and apoptosis of HH cells. IFN-α combined with matrine could significantly increase the expressions of Fas, Caspase8, Caspase3 and Bax, and significantly inhibit the expression of Bcl-2 But had no effect on Fas L expression. Conclusion IFN-α plays a therapeutic role in CTCL by inhibiting cell proliferation and promoting apoptosis. Compared with IFN-α alone, IFN-α combined with matrine can significantly improve the therapeutic effect of CTCL and reduce the amount of IFN-α.