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目的探讨中药活性成分蛇床子素对抗肿瘤药物TRAIL抗乳腺癌活性的影响并研究其机制。方法用蛇床子素联合TRAIL体外治疗乳腺癌细胞系BT-20,MTT法检测肿瘤细胞的细胞活力,Annexin V/PI染色检测肿瘤细胞的凋亡,免疫共沉淀法检测RIP1-FADD-caspase-8复合物的形成。Western blot法检测BT-20细胞caspase-8的活化及蛇床子素对BT-20细胞c IAP2蛋白表达的影响。结果联用蛇床子素显著提高TRAIL对BT-20细胞活力的抑制率和凋亡诱导活性。免疫共沉淀及Western blot结果发现联合蛇床子素后,TRAIL治疗的BT-20细胞内的RIP1-FADD-caspase-8复合物水平及caspase-8的活化程度显著提高。Western blot结果发现蛇床子素对BT-20细胞内的c IAP2蛋白表达有抑制作用。在BT-20细胞中转染c IAP2表达质粒后,蛇床子素对TRAIL抗肿瘤活性的促进作用受到抑制。结论蛇床子素通过促进死亡受体复合物的形成增强TRAIL对乳腺癌细胞的凋亡诱导效应。
Objective To investigate the effect of the active component osthole on the anti-tumor activity of TRAIL and its mechanism. Methods The viability of tumor cells was determined by MTT assay. The apoptosis of tumor cells was detected by Annexin V / PI staining. The expression of RIP1-FADD-caspase-8 was detected by co-immunoprecipitation Complex formation. Western blot was used to detect the activation of caspase-8 in BT-20 cells and the effect of osthole on c IAP2 protein expression in BT-20 cells. Results Combined with osthole, the inhibitory rate and apoptosis-inducing activity of TRAIL on the viability of BT-20 cells were significantly increased. Co-immunoprecipitation and Western blot showed that the level of RIP1-FADD-caspase-8 complex and the activation of caspase-8 in BT-20 cells treated with TRAIL were significantly increased after combined with osthole. Western blot results showed that osthole could inhibit c IAP2 protein expression in BT-20 cells. After transfection of c IAP2 expression plasmid in BT-20 cells, osthole could inhibit the antitumor activity of TRAIL. Conclusion Osthole could enhance the apoptosis-inducing effect of TRAIL on breast cancer cells by promoting the formation of death receptor complex.