Lab of Stem Cell and Tissue Engineering, Department of Histology and Embryology, Chongqing Medical University, Chongqing 400016, China.
Department of Neurology, Yongchuan Hospital of Chongqing Medical University, Chongqing 400016, China.
Life Sci. 2020 Jun 15;251:117424. doi: 10.1016/j.lfs.2020.117424. Epub 2020 Feb 11.
Dysfunction of the Hippo-Yes-Associated Protein (YAP) signaling pathway is known to be associated with hepatocellular carcinoma (HCC). Evodiamine (Evo), a plant-derived bioactive alkaloid, exerts inhibitory effects on cancer. However, the precise influence of Evo on HCC and its potential effects on Hippo-YAP signaling have yet to be ascertained. Here, the effects of Evo on cell proliferation and apoptosis were evaluated using HCC cell lines (HepG2 and Bel-7402) and nude mice with xenograft tumors. We further investigated whether Evo exerts anti-HCC activity through effects on Hippo-YAP signaling in vitro with the aid of XMU-MP-1, an inhibitor of the key component of this pathway, mammalian sterile 20-like kinase 1/2.
Cell proliferation and apoptosis were assessed using 5-ethynyl-2'-deoxyuridine staining, colony formation, flow cytometry, hematoxylin-eosin and dUTP nick-end labeling experiments. Bioinformatics and real-time quantitative polymerase chain reaction (RT-qPCR) arrays were performed to determine the associations among Evo, HCC progression and the Hippo-YAP pathway. The expression patterns of components of Hippo-YAP signaling and apoptotic genes were further examined via RT-qPCR and immunoblotting.
Evo inhibited proliferation and promoted apoptosis of HCC cell lines in vitro, and attenuated xenograft tumor formation in nude mice in vivo. Mechanistically, Evo treatment stimulated the Hippo-YAP signaling pathway. In vitro, the effects of Evo on HCC cell proliferation and apoptosis were alleviated by XMU-MP-1.
Our collective results revealed that the anti-HCC effects of Evo were correlated with the Hippo-YAP signaling pathway.
已知 Hippo-Yes 相关蛋白 (YAP) 信号通路功能障碍与肝细胞癌 (HCC) 有关。吴茱萸碱 (Evo) 是一种植物源性生物活性生物碱，对癌症具有抑制作用。然而，Evo 对 HCC 的确切影响及其对 Hippo-YAP 信号通路的潜在影响尚未确定。在这里，我们使用 HCC 细胞系 (HepG2 和 Bel-7402) 和裸鼠异种移植瘤评估了 Evo 对细胞增殖和凋亡的影响。我们进一步研究了 Evo 是否通过 Hippo-YAP 信号通路在体外对 HCC 发挥抗活性，使用了该通路关键成分的抑制剂 XMU-MP-1，哺乳动物 sterile 20 样激酶 1/2。
使用 5-乙炔基-2'-脱氧尿苷染色、集落形成、流式细胞术、苏木精-伊红和 dUTP 末端标记实验评估细胞增殖和凋亡。生物信息学和实时定量聚合酶链反应 (RT-qPCR) 阵列用于确定 Evo、HCC 进展和 Hippo-YAP 通路之间的关联。通过 RT-qPCR 和免疫印迹进一步检查 Hippo-YAP 信号和凋亡基因的组成部分的表达模式。
Evo 抑制 HCC 细胞系的体外增殖并促进其凋亡，并减轻体内裸鼠异种移植瘤的形成。在机制上，Evo 处理刺激 Hippo-YAP 信号通路。在体外，Evo 对 HCC 细胞增殖和凋亡的影响通过 XMU-MP-1 减轻。
我们的综合结果表明，Evo 的抗 HCC 作用与 Hippo-YAP 信号通路有关。
