吲哚胺2,3-双加氧酶1抑制剂Epacadostat联合γ干扰素诱导三阴乳腺癌细胞系凋亡

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (5): 615-620.

吲哚胺2,3-双加氧酶1抑制剂Epacadostat联合γ干扰素诱导三阴乳腺癌细胞系凋亡

胡庆庆, 鲁兖^*

浙江大学医学院附属第四医院药剂科, 浙江金华 322023

收稿日期:2019-08-15 修回日期:2020-01-02 出版日期:2020-05-05 发布日期:2020-04-30
通讯作者: *2504159@zju.edu.cn
基金资助:
国家自然科学基金(81502473)

Indoleamine 2,3-dioxygenase 1 inhibitor Epacadostat combined with IFN-γ induces apoptosis of triple-negative breast cancer cell lines

HU Qing-qing, LU Yan^*

Department of Pharmacy, the Fourth Affiliated Hospital, School of Medicine, Zhejiang University, Jinhua 322023, China

Received:2019-08-15 Revised:2020-01-02 Online:2020-05-05 Published:2020-04-30
Contact: *2504159@zju.edu.cn

摘要/Abstract

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。

关键词: 三阴乳腺癌, γ干扰素, 吲哚胺2,3-双加氧酶1

Abstract: Objective To investigate the effect of IFN-γ combined 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-γ combined 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-γ showed a weaker killing effect on triple-negative breast cancer cells; IFN-γ significantly 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-γ for 48 h was also significantly increased; IFN-γ combined 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-γ on triple-negative breast cancer.

Key words: triple-negative breast cancer, IFN-γ, indoleamine 2,3-dioxygenase 1

中图分类号:

R737.9

胡庆庆, 鲁兖. 吲哚胺2,3-双加氧酶1抑制剂Epacadostat联合γ干扰素诱导三阴乳腺癌细胞系凋亡[J]. 基础医学与临床, 2020, 40(5): 615-620.

HU Qing-qing, LU Yan. Indoleamine 2,3-dioxygenase 1 inhibitor Epacadostat combined with IFN-γ induces apoptosis of triple-negative breast cancer cell lines[J]. Basic & Clinical Medicine, 2020, 40(5): 615-620.

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