Department of Pharmaceutical Biology, Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University, Staudinger Weg 5, 55128, Mainz, Germany.
Department of Molecular Biology, Faculty of Medical Laboratory Sciences, Al-Neelain University, Khartoum, Sudan.
Invest New Drugs. 2021 Dec;39(6):1523-1537. doi: 10.1007/s10637-021-01137-y. Epub 2021 Jul 2.
Background Triptolide is an active natural product, which inhibits cell proliferation, induces cell apoptosis, suppresses tumor metastasis and improves the effect of other therapeutic treatments in several cancer cell lines by affecting multiple molecules and signaling pathways, such as caspases, heat-shock proteins, DNA damage and NF-ĸB. Purpose We investigated the effect of triptolide towards NF-ĸB and GATA1. Methods We used cell viability assay, compare and cluster analyses of microarray-based mRNA transcriptome-wide expression data, gene promoter binding motif analysis, molecular docking, Ingenuity pathway analysis, NF-ĸB reporter cell assay, and electrophoretic mobility shift assay (EMSA) of GATA1. Results Triptolide inhibited the growth of drug-sensitive (CCRF-CEM, U87.MG) and drug-resistant cell lines (CEM/ADR5000, U87.MGΔEGFR). Hierarchical cluster analysis showed six major clusters in dendrogram. The sensitive and resistant cell lines were statistically significant (p = 0.65 × 10) distributed. The binding motifs of NF-κB (Rel) and of GATA1 proteins were significantly enriched in regions of 25 kb upstream promoter of all genes. IPA showed the networks, biological functions, and canonical pathways influencing the activity of triptolide towards tumor cells. Interestingly, upstream analysis for the 40 genes identified by compare analysis revealed ZFPM1 (friend of GATA protein 1) as top transcription regulator. However, we did not observe any effect of triptolide to the binding of GATA1 in vitro. We confirmed that triptolide inhibited NF-κB activity, and it strongly bound to the pharmacophores of IκB kinase β and NF-κB in silico. Conclusion Triptolide showed promising inhibitory effect toward NF-κB, making it a potential candidate for targeting NF-κB.
雷公藤红素是一种活性天然产物，通过影响多种分子和信号通路，如半胱天冬酶、热休克蛋白、DNA 损伤和 NF-κB，抑制细胞增殖，诱导细胞凋亡，抑制肿瘤转移，并提高几种癌细胞系中其他治疗方法的疗效。
研究雷公藤红素对 NF-κB 和 GATA1 的影响。
采用细胞活力测定法、基于微阵列的 mRNA 转录组全表达数据的比较和聚类分析、基因启动子结合基序分析、分子对接、Ingenuity 通路分析、NF-κB 报告细胞检测和 GATA1 的电泳迁移率变动分析（EMSA）。
雷公藤红素抑制了药物敏感（CCRF-CEM、U87.MG）和耐药细胞系（CEM/ADR5000、U87.MGΔEGFR）的生长。层次聚类分析显示树状图中有六个主要聚类。敏感和耐药细胞系在统计学上有显著差异（p=0.65×10）。NF-κB（Rel）和 GATA1 蛋白的结合基序在所有基因的 25kb 上游启动子区域显著富集。IPA 显示了影响雷公藤红素对肿瘤细胞活性的网络、生物学功能和经典途径。有趣的是，比较分析确定的 40 个基因的上游分析显示 ZFPM1（GATA 蛋白 1 的朋友）是顶级转录调节因子。然而，我们没有观察到雷公藤红素对 GATA1 结合的任何体外影响。我们证实雷公藤红素抑制 NF-κB 活性，并且它在计算机中强烈结合 IκB 激酶β和 NF-κB 的药效团。
雷公藤红素对 NF-κB 表现出有希望的抑制作用，使其成为靶向 NF-κB 的潜在候选药物。
