Triptonide acts as a novel antiprostate cancer agent mainly through inhibition of mTOR signaling pathway
Triptonide acts as a novel antiprostate cancer agent mainly through inhibition of mTOR signaling pathway
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Triptonide作为一种新型抗前列腺癌药物主要通过抑制mTOR信号通路发挥作用
DOI：
10.1002/pros.23834
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发表时间：
2019-08-01
期刊：
影响因子：
2.8
通讯作者：
Jiang, Ming
中科院分区：
文献类型：
--
作者：
Dong, Fulu;Yang, Ping;Jiang, Ming
背景前列腺癌（PCa）发病率的不断上升，迫切需要开发新的有效治疗PCa的药物。通过对中草药的筛选，发现雷公藤内酯酮具有较强的抗癌活性。本研究旨在探讨雷公藤内酯醇对前列腺癌细胞抗前列腺癌活性的影响及其机制。方法用不同浓度（0、1.25、2.5、5、10、20、40、80、160和320 nM）的雷公藤内酯酮分别处理3株人晚期前列腺癌细胞系PC 3、DU 145和LNCap以及1株人正常前列腺上皮细胞系RWPE 72 h。然后，通过细胞计数试剂盒-8评估细胞活力。用不同剂量（0-20 nM）的雷公藤内酯醇处理PCa细胞72小时。流式细胞仪检测细胞周期和凋亡。携带人PCa异种移植物的裸鼠每天腹腔注射雷公藤内酯酮（10 mg/kg/d）或磷酸盐缓冲盐水作为对照，持续35天。通过Illumina Hiseq测序平台进行RNA测序（RNA-seq），并通过实时聚合酶链反应确认。使用基因本体论、京都基因和基因组百科全书途径分析和独创性途径分析来分析RNA-seq结果。结果雷公藤内酯酮对人前列腺癌细胞PC 3、DU 145和LNCap的体外增殖均有抑制作用，其IC 50值分别为11.961、10.259和12.012 nM。雷公藤内酯酮（10 mg/kg）在体内有效地抑制PCa细胞异种移植物的生长，抑制率超过97.95%。雷公藤内酯酮（5 nM）治疗显着促进细胞凋亡和保留细胞周期停滞在G2/M期。RNA-seq数据显示，共有936个基因在雷公藤内酯醇治疗中上调或下调。此外，雷帕霉素的机械靶点（mTOR）和下游蛋白p70 S6 K的磷酸化都受到抑制，在PCa细胞中最明显。结论雷公藤内酯酮可通过抑制mTOR磷酸化及下游相关信号通路的活性，有效抑制PCa的生长。
Background The increasing incidence of prostate cancer (PCa) indicates an urgent need for the development of new effective drugs in PCa therapy. Triptonide has been reported to have a strong inhibition activity in cancers through screening of Chinese herbal medicine. This study aims to investigate the effects of triptonide on anti-PCa activity and its mechanisms. Methods Three human advanced PCa cell lines PC3, DU145, and LNCap, and a human normal prostate epithelial cell line RWPE were treated with a range (0, 1.25, 2.5, 5, 10, 20, 40, 80, 160, and 320 nM) of triptonide concentrations for 72 hours respectively. Then, cell viability was assessed by cell counting kit-8. PCa cells were treated with different doses (0-20 nM) of triptonide for 72 hours. Cell cycle and apoptosis were assessed by flow cytometry assays. Nude mice bearing human PCa xenografts were intraperitoneally injected daily with either triptonide (10 mg/kg/d) or phosphate-buffered saline as a control for 35 days. RNA-sequencing (RNA-seq) was performed by an Illumina Hiseq Sequencing platform and confirmed by a real-time polymerase chain reaction. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and ingenuity pathway analysis were used to analyze RNA-seq results. Results Triptonide effectively inhibits the proliferation of human PCa cells PC3, DU145, and LNCap in vitro with their IC50 values as 11.961, 10.259, and 12.012 nM, respectively. Triptonide (10 mg/kg) potently inhibits the growth of PCa cell xenografts in vivo at an inhibition rate of over 97.95%. Treatment with triptonide (5 nM) significantly promotes cell apoptosis and retaining cell-cycle arrest in the G2/M phase. RNA-seq data revealed that total of 936 genes were upregulated or downregulated in triptonide treated. Moreover, the phosphorylation of mechanistic target of rapamycin (mTOR) and the downstream protein p70S6K were both inhibited, most obviously in PCa cells. Conclusions Our findings suggest that triptonide can efficaciously suppress PCa growth in vitro and in vivo via inhibiting the phosphorylation of mTOR and the activities of related downstream signaling pathways.