Backgrounds: Abnormal metabolism is the hallmark of hepatocellular carcinoma. Targeting energy metabolism has become the major focus of cancer therapy. The natural product, sanguinarine, displays remarkable anti-tumor properties by disturbing energy homeostasis; however, the underlying mechanism has not yet been elucidated. Methods: The anticancer activity of sanguinarine was determined using CCK-8 and colony formation assay. Morphological changes of induced cell death were observed under electron microscopy. Necroptosis and apoptosis related markers were detected using western blotting. PKM2 was identified as the target by transcriptome sequencing. Molecular docking assay was used to evaluate the binding affinity of sanguinarine to the PKM2 molecule. Furthermore, Alb-CreERT2; PKM2loxp/loxp; Rosa26RFPmice was used to construct the model of HCC—through the intervention of sanguinarine in vitro and in vivo—to accurately explore the regulation effect of sanguinarine on cancer energy metabolism. Results: Sanguinarine inhibited tumor proliferation, metastasis and induced two modes of cell death. Molecular docking of sanguinarine with PKM2 showed appreciable binding affinity. PKM2 kinase activity and aerobic glycolysis rate declined, and mitochondrial oxidative phosphorylation was inhibited by sanguinarine application; these changes result in energy deficits and lead to necroptosis. Additionally, sanguinarine treatment prevents the translocation of PKM2 into the nucleus and suppresses the interaction of PKM2 with β-catenin; the transcriptional activity of PKM2/β-catenin signaling and its downstream genes were decreased. Conclusions: Sanguinarine showed remarkable anti-HCC activity via regulating energy metabolism by PKM2/β-catenin signaling. On the basis of these investigations, we propose that sanguinarine might be considered as a promising compound for discovery of anti-HCC drugs.
背景：代谢异常是肝细胞癌的典型特征。靶向能量代谢已成为癌症治疗的主要焦点。天然产物血根碱通过干扰能量稳态展现出显著的抗肿瘤特性，但其潜在机制尚未阐明。方法：采用CCK-8法和克隆形成实验测定血根碱的抗癌活性。通过电子显微镜观察诱导细胞死亡的形态学变化，并使用蛋白质印迹法检测坏死性凋亡和凋亡相关标志物。通过转录组测序鉴定PKM2为作用靶点，采用分子对接实验评估血根碱与PKM2分子的结合亲和力。进一步利用Alb-CreERT2; PKM2loxp/loxp; Rosa26RFP小鼠构建肝细胞癌模型，通过体内外血根碱干预，精准探究其对肿瘤能量代谢的调控作用。结果：血根碱能抑制肿瘤增殖与转移，并诱导两种细胞死亡模式。血根碱与PKM2的分子对接显示良好结合亲和力。血根碱干预可降低PKM2激酶活性及有氧糖酵解速率，抑制线粒体氧化磷酸化，导致能量缺失并引发坏死性凋亡。此外，血根碱处理能阻止PKM2向细胞核转位，抑制PKM2与β-连环蛋白的相互作用，从而降低PKM2/β-连环蛋白信号通路及其下游基因的转录活性。结论：血根碱通过调控PKM2/β-连环蛋白信号通路介导的能量代谢，展现出显著的抗肝细胞癌活性。基于本研究，我们认为血根碱可作为抗肝细胞癌药物研发的潜在候选化合物。
Sanguinarine Induces Necroptosis of HCC by Targeting PKM2 Mediated Energy Metabolism
肿瘤(癌症)患者之家