从Menispermum dauricum DC提取的N-去甲基达乌林素通过下调NF-κB信号通路抑制三阴性乳腺癌在二维和三维模型中的生长

N-desmethyldauricine from Menispermum dauricum DC suppresses triple-negative breast cancer growth in 2D and 3D models by downregulating the NF-κB signaling pathway

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CHEMICO-BIOLOGICAL INTERACTIONS

影响因子:5.4

分区:医学2区 / 毒理学1区生化与分子生物学2区药学2区

发表日期:2024 Aug 01

作者: Wenting Liu, Yan Yu, Tao Hou, Hongli Wei, Fangbin Lv, Aijin Shen, Yanfang Liu, Jixia Wang, Dongmei Fu

DOI: 10.1016/j.cbi.2024.111113

keywords: 3D tumor spheroids; M. dauricum; N-desmethyldauricine; NF-κB; TNBC

摘要

三阴性乳腺癌（TNBC）是乳腺癌中最具侵袭性的亚型，缺乏靶向治疗方案。传统中药Menispermum dauricum DC（M. dauricum）及其化合物已被报道具有抗肿瘤活性，但其抗TNBC的作用尚未明确。本研究采用多组分生物活性和结构引导的方法，分离鉴定了M. dauricum中的达乌林素和N-去甲基达乌林素，发现其具有抗TNBC作用。细胞计数试剂盒8（CCK-8）实验证明，达乌林素和N-去甲基达乌林素抑制了四种TNBC细胞系的增殖，半数最大抑制浓度（IC50）范围为5.01 μM至13.16 μM。进一步研究显示，N-去甲基达乌林素诱导细胞凋亡，阻滞细胞周期在G0/G1期，并抑制细胞迁移。Western blot分析显示促凋亡蛋白裂解的多腺苷二磷酸核糖聚合酶1（cleaved-PARP1）上调，G0/G1期相关蛋白周期蛋白依赖性激酶2（CDK2）和周期蛋白D1（Cyclin D1）下调，迁移相关蛋白基质金属蛋白酶9（MMP-9）下调。此外，N-去甲基达乌林素还降低了核因子κB（NF-κB）重要亚基p65的蛋白表达。三维肿瘤球体的抗增殖实验表明，N-去甲基达乌林素减弱了细胞间粘附并抑制了TNBC三维球体的生长。综上，N-去甲基达乌林素可抑制TNBC细胞的增殖，降低NF-κB通路中p65的表达。

Abstract

Triple negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, for which targeted therapy regimens are lacking. The traditional Chinese medicine Menispermum dauricum DC (M. dauricum) and its compounds have been reported to have antitumor activity against various cancers; however, their anti-TNBC activity is unknown. In this work, dauricine and N-desmethyldauricine from M. dauricum were separated and identified to have anti-TNBC via a multi-component bioactivity and structure-guided method. The cell counting kit 8 assay showed that dauricine and N-desmethyldauricine inhibited the proliferation of four tested TNBC cell lines, with half maximal inhibitory concentration values ranging from 5.01 μM to 13.16 μM. Further research suggested that N-desmethyldauricine induced cell apoptosis, arrested cell cycle progression in the G0/G1 phase, and inhibited cell migration. Western blot analysis revealed that the proapoptotic protein cleaved-poly-ADP-ribose polymerase 1 was upregulated, and the G0/G1 phase-related proteins cyclin-dependent kinase 2 and cyclin D1 and the migration-related protein matrix metallopeptidase 9 were downregulated. Furthermore, N-desmethyldauricine decreased the protein expression of p65, an important subunit of nuclear factor kappa-beta (NF-κB). Moreover, an antiproliferation assay of three-dimensional (3D) tumor spheroids showed that N-desmethyldauricine diminished cell‒cell adhesion and suppressed the growth of TNBC 3D spheroids. Taken together, these findings indicate that N-desmethyldauricine inhibited the proliferation of TNBC cells and decreased the expression of p65 in the NF-κB pathway.