摘要
目的探讨γ干扰素(IFN-γ)与吲哚胺2,3-双加氧酶1(IDO1)抑制剂Epacadostat(Epa)联用对三阴乳腺癌细胞系4T1和MDA-MB-231凋亡作用的影响。方法IFN-γ与Epa单独或联合处理4T1和MDA-MB-231细胞48 h,流式细胞仪检测细胞凋亡;蛋白免疫印迹检测IFN-γ处理后细胞IDO1分子的表达;ELISA检测培养上清中犬尿氨酸的含量;CRISPR-Cas9法敲除IDO1分子,蛋白免疫印迹检测敲除效率。结果IFN-γ诱导三阴乳腺癌细胞系凋亡的作用较弱;IFN-γ可显著上调三阴乳腺癌细胞系IDO1的表达(P<0.001),而且细胞内Kyn的水平也显著上调(P<0.01);敲除IDO1后IFN-γ对4T1和MDA-MB-231细胞的凋亡作用大大增强;IFN-γ联用Epa促进了4T1和MDA-MB-231细胞的凋亡,大大延长了荷瘤小鼠的生存时间。结论IDO1抑制剂Epa可大大增强IFN-γ对三阴乳腺癌的治疗作用。
Objective To investigate the effect of IFN-γcombined with IDO1 inhibitor Epacadostat(Epa)on the treatment of triple-negative breast cancer cell lines 4T1 and MDA-MB-231.Methods Following treatment with IFN-γ,Epa or IFN-γcombined with Epa on 4T1 and MDA-MB-231 cells for 48 h,the apoptosis of cells in each group was detected by flow cytometry.The expression of IDO1 was detected by Western blot and the yield of Kyn in 4T1or MDA-MB-231 cell lysate was detected by ELISA.The IDO1 was knocked out by using CRISPR-Cas9 system and knockout efficiency was examined by Western blot.Results IFN-γshowed a weaker killing effect on triple-negative breast cancer cells;IFN-γsignificantly up-regulated the expression of IDO1 in triple-negative breast cancer cell lines(P<0.001),and the level of Kyn in cell lysate was also significantly up-regulated(P<0.01);cell apoptosis of IDO1 knock-out 4T1(P<0.01)or MDA-MB-231(P<0.01)cells treated with IFN-γfor 48 h was also significantly increased;IFN-γcombined with IDO1 inhibitor Epa promoted the apoptosis of 4T1 and MDA-MB-231 cells,and also prolonged the survival time of tumor-bearing mice.Conclusions The IDO1 inhibitor Epa might significantly enhance the therapeutic effect of IFN-γon triple-negative breast cancer.
作者
胡庆庆
鲁兖
HU Qing-qing;LU Yan(Department of Pharmacy, the Fourth Affiliated Hospital, School of Medicine, Zhejiang University, Jinhua 322023, China)
出处
《基础医学与临床》
CSCD
2020年第5期615-620,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(81502473)。
作者简介
通讯作者:鲁兖,Email:2504159@zju.edu.cn。
