药学学报, 2017, 52(9): 1416-1423
引用本文:
庞伟强, 高磊, 窦玥莹, 刘璐, 张娜, 刘晓嘉, 刘阳, 杨兆勇, 宋丹青, 邓洪斌. IDO1天然小分子抑制剂的筛选及抗肿瘤作用研究[J]. 药学学报, 2017, 52(9): 1416-1423.
PANG Wei-qiang, GAO Lei, DOU Yue-ying, LIU Lu, ZHANG Na, LIU Xiao-jia, LIU Yang, YANG Zhao-yong, SONG Dan-qing, DENG Hong-bin. Screening of natural small molecule IDO-1 inhibitors with anti-tumor role[J]. Acta Pharmaceutica Sinica, 2017, 52(9): 1416-1423.

IDO1天然小分子抑制剂的筛选及抗肿瘤作用研究
庞伟强1, 高磊2, 窦玥莹1, 刘璐1, 张娜1, 刘晓嘉1, 刘阳1, 杨兆勇1, 宋丹青1, 邓洪斌1
1. 中国医学科学院、北京协和医学院医药生物技术研究所, 北京 100050;
2. 吉林省药品检验所, 吉林 长春 130000
摘要:
PCR扩增人吲哚胺2,3-双加氧酶1(indoleamine 2,3-dioxygenase 1,IDO1)基因启动子上游区域1 245 bp基因片段,将其插入到pGL4.20-basic载体中构建了pGL4-IDO1-luc荧光素酶重组质粒。基于双荧光素酶报告基因方法建立IDO1抑制剂筛选模型,筛选能够下调肿瘤细胞中IDO1表达的天然活性小分子化合物。采用MTT、Western blotting和乳酸脱氢酶(LDH)等方法探讨阳性化合物的抗肿瘤作用及其对IDO1的调控机制。化合物川楝素(toosendanin,NS-180)能显著下调IFN-γ诱导的肿瘤细胞中IDO1表达。在A549细胞中,NS-180可抑制STAT1和STAT3的磷酸化,从而下调IFN-γ诱导的IDO1蛋白表达。LDH释放实验表明,NS-180可促进NK细胞对A549细胞的杀伤作用。综上所述,筛选获得的天然小分子NS-180是一类新型高效的IDO1抑制剂,可能成为靶向IDO1的肿瘤免疫治疗候选药物。
关键词:   
Screening of natural small molecule IDO-1 inhibitors with anti-tumor role
PANG Wei-qiang1, GAO Lei2, DOU Yue-ying1, LIU Lu1, ZHANG Na1, LIU Xiao-jia1, LIU Yang1, YANG Zhao-yong1, SONG Dan-qing1, DENG Hong-bin1
1. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China;
2. Jilin Institute for Drug Control, Changchun 130000, China
Abstract:
Fragments of the human indoleamine 2,3-dioxygenase 1 (IDO1) gene 5'-UTR (untranslated 1 245 bp region) promoters were amplified by PCR and cloned into pGL4.20 vector in the construction of reporter vector pGL4-IDO1-luc. A549 cells were transfected with the constructed plasmid and IDO1 inhibitor screening model was established with dual-luciferase reporter assay. Based on the model, we screened natural small molecules which could down-regulate the expression of IDO1 on tumor cells. The anti-tumor activities were examined by MTT, Western blotting and lactic dehydrogenase (LDH) release assays. Toosendanin (NS-180) down regulated the IDO1 expression and inhibited IFN-γ-induced STAT1 and STAT3 phosphorylation in A549 cells. Moreover, NS-180 significantly increased the cytotoxicity of co-cultured NK cells on A549 cells in LDH release assays. In summary, NS-180 is a novel and potent IDO1 inhibitor, which has an antitumor activity for cancer immunotherapies.
Key words:   
收稿日期: 2017-05-22
DOI: 10.16438/j.0513-4870.2017-0506
基金项目: 国家自然科学基金面上资助项目(81473248);中国医学科学院重大协同创新项目-重大前沿研究(2016-12M-1-011).
通讯作者: 邓洪斌,Tel:86-10-63169876,Fax:86-10-63017302,E-mail:hdeng@imb.pumc.edu.cn
Email: hdeng@imb.pumc.edu.cn
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