IDO1天然小分子抑制剂的筛选及抗肿瘤作用研究
发布时间：2018-10-12 12:54
【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p GL4.20-basic载体中构建了p GL4-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。
[Abstract]:The 1 245 bp gene fragment of the upstream region of the promoter of the human indoleamine 23-dioxygenase 1 (IDO1) gene was amplified by PCR and inserted into the p GL4.20-basic vector to construct the recombinant plasmid of p GL4-IDO1-luc luciferase. A screening model of IDO1 inhibitor was established based on double luciferase reporter gene method to screen natural active small molecular compounds which can down-regulate the expression of IDO1 in tumor cells. MTT,Western blotting and lactate dehydrogenase (LDH) were used to investigate the antitumor effect of positive compounds and its regulatory mechanism on IDO1. Toosendanin (toosendanin,NS-180) could significantly down-regulate the expression of IDO1 in tumor cells induced by IFN- 纬. In A549 cells, NS-180 inhibited the phosphorylation of STAT1 and STAT3 and down-regulated the expression of IDO1 protein induced by IFN- 纬. LDH release assay showed that NS-180 could promote the cytotoxicity of NK cells to A549 cells. In conclusion, the obtained natural small molecule NS-180 is a novel and efficient IDO1 inhibitor, which may be a candidate for tumor immunotherapy targeting IDO1.
【作者单位】： 中国医学科学院北京协和医学院医药生物技术研究所;吉林省药品检验所;
【基金】：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)
【分类号】：R73-36
[Abstract]:The 1 245 bp gene fragment of the upstream region of the promoter of the human indoleamine 23-dioxygenase 1 (IDO1) gene was amplified by PCR and inserted into the p GL4.20-basic vector to construct the recombinant plasmid of p GL4-IDO1-luc luciferase. A screening model of IDO1 inhibitor was established based on double luciferase reporter gene method to screen natural active small molecular compounds which can down-regulate the expression of IDO1 in tumor cells. MTT,Western blotting and lactate dehydrogenase (LDH) were used to investigate the antitumor effect of positive compounds and its regulatory mechanism on IDO1. Toosendanin (toosendanin,NS-180) could significantly down-regulate the expression of IDO1 in tumor cells induced by IFN- 纬. In A549 cells, NS-180 inhibited the phosphorylation of STAT1 and STAT3 and down-regulated the expression of IDO1 protein induced by IFN- 纬. LDH release assay showed that NS-180 could promote the cytotoxicity of NK cells to A549 cells. In conclusion, the obtained natural small molecule NS-180 is a novel and efficient IDO1 inhibitor, which may be a candidate for tumor immunotherapy targeting IDO1.
【作者单位】： 中国医学科学院北京协和医学院医药生物技术研究所;吉林省药品检验所;
【基金】：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)
【分类号】：R73-36
【相似文献】
相关期刊论文 前6条
1 王俊;郭燕;章必成;关华军;陈正堂;;VEGF-C基因3′端未翻译区对荧光素酶表达的影响[J];解放军医学杂志;2007年09期
2 孟希亭;林晨;梅佳;王海娟;马飞;张金龙;张颖;钱海利;;活体成像系统检测携荧光素酶增殖缺陷型腺病毒在小鼠体内的表达和分布[J];中国肿瘤生物治疗杂志;2011年04期
3 张绘宇;郑国兵;张金栋;刘启才;;利用荧光素酶标记的肿瘤细胞建立活体成像动物模型[J];中国体视学与图像分析;2013年01期
4 张金栋;吕景礼;周问渠;刘启才;李晓艳;;整合荧光素酶的稳定细胞克隆建立裸鼠肺癌静脉移植瘤模型的探索[J];实用医学杂志;2012年07期
5 吴学明;;miR-22在卵巢癌中的表达及其分子机制研究[J];中国妇幼保健;2014年23期
6 徐元基;周涛;杜芝燕;刘刚;巩伟丽;于晓Y，
本文编号：2266188
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2266188.html
