双硫仑联合Cu对淋巴瘤Raji细胞增殖与凋亡影响及其机制的探讨

目的:研究双硫仑(Disulfiram,DS)联合Cu离子对Burkitt’s淋巴瘤细胞株(Raji细胞)增殖和凋亡的影响及其与JNK和NF-κB通路的关系。方法:不同浓度DS和DS/CuMTT法检测对Raji细胞的增殖抑制作用;AnnexinⅤ-FITC/PI流式细胞术检测不同浓度DS和DS/Cu作用Raji细胞后凋亡细胞比例;蛋白质印迹法检测Cu、DS及DS/Cu处理细胞后p65、p-JNK及c-jun蛋白表达的变化。结果:DS和DS/Cu对Raji细胞均具有抑制增殖作用,作用72h两组IC50值分别为(0.793±0.08)和(0.085±0.015)μmol/mL,DS/Cu对Raji细胞增殖抑制作用显著强于DS单药,P=0.000。DS单药及DS/Cu对Raji细胞均有诱导凋亡作用,但DS/Cu诱导Raji细胞凋亡比例显著高于DS单药,P=0.001。蛋白质印迹法显示DS及DS/Cu作用Raji细胞后p-JNK及c-jun蛋白表达水平均显著增加、p65蛋白表达水平显著下调。结论:DS/Cu及DS对Raji细胞均有抑制增殖及诱导凋亡作用,其中DS/Cu的作用显著强于DS单药,其机制与同时抑制NF-κB和活化JNK通路有关。

Inhibition effect of Disulfiram and combined with Cu on the proliferation and apopsis of Raji cells and its medchanism

OBJECTIVE: To investigate the proliferation and apoptosis of Disulfiram(DS) and DS/Cu on Raji cells in Burkitt’s lymphoma and the relationship with JNK and NF-κB pathway.METHODS: The Raji cells were treated with various concentrations of DS and DS/Cu.The inhibitory ratio was measured by MTT assay.with flow cytometric the apoptotic Raji cells were analyzed after treatment with DS and DS/Cu.Western bloting were used to analyzed p65,p-JNK and c-jun expression in Raji cells after treatment with Cu,DS and DS/Cu.RESULTS: Different concentrations of DS could inhibit the proliferation of Raji cells with the 50% inhibitory concentration(IC50) values of(0.793±0.08) μmol/mL.In combination with Cu,the IC50 of DS to Raji cells was significantly reduced to(0.085±0.015) μmol/mL(P=0.000).The flow cytometry result showed significant incrementing of apoptotic Raji cells after treatment with DS and DS/Cu,DS/Cu had a significantly higher proportion of apoptotic Raji cells than DS(P=0.001).Western blot results showed markedly increase the expression of p-JNK,c-jun and decrease the expression of p65 in Raji cells treated with a DS and DS/Cu.CONCLUSIONS: DS and DS/Cu can inhibit the proliferation and induce the apopsis of Raji cells.This may be caused by simultaneous activation of JNK pathway and inhibition of NF-κB pathway.