Division of Gastroenterology and Hepatology, Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei City 110, Taiwan; Division of Gastroenterology and Hepatology, Department of Internal Medicine, Taipei Medical University-Shuang Ho Hospital, New Taipei City 235, Taiwan.
Department of Emergency Medicine, Taipei Medical University-Shuang Ho Hospital, New Taipei City 235, Taiwan; Graduate Institute of Injury Prevention and Control, College of Public Health, Taipei Medical University, Taipei City 110, Taiwan; Department of Emergency Medicine, School of Medicine, Taipei Medical University, Taipei City 110, Taiwan.
Phytomedicine. 2023 Jan;108:154478. doi: 10.1016/j.phymed.2022.154478. Epub 2022 Sep 25.
Activation of mitogen-activated protein kinase (MAPK) and PI3K signaling confers resistance against sorafenib, a mainstay treatment for advanced hepatocellular carcinoma (HCC). Antrocin and ovatodiolide constitute as the most potent secondary metabolites isolated from Antrodia camphorata and Anisomeles indica, respectively. Both natural compounds have recently gained a lot of attention due to their putative inhibition of MAPK and PI3K signaling in various solid cancers. However, whether their combination is effective in HCC remains unknown. Here, we investigated their effect, alone or in various combinations, on MAPK and PI3K signaling pathways in HCC cells. An array of in vitro study were used to investigate anticancer and stemness effects to treat HCC, such as cytotoxicity, drug combination index, migration, invasion, colony formation, and tumor sphere formation. Drug effect in vivo was evaluated using mouse xenograft models. In this study, antrocin and ovatodiolide synergistically inhibited the SNU387, Hep3B, Mahlavu, and Huh7 cell lines. Sequential combination treatment of Huh7 and Mahlavu with ovatodiolide followed by antrocin resulted stronger cytotoxic effect than did treatment with antrocin followed by ovatodiolide, their simultaneous administration, antrocin alone, or ovatodiolide alone. In the Huh7 and Mahlavu cell lines, ovatodiolide→antrocin significantly suppressed colony formation and proliferation as well as markedly downregulated ERK1/2, Akt, and mTOR expression. Inhibition of ERK1/2 and Akt/mTOR signaling by ovatodiolide→antrocin suppressed ribosomal biogenesis, autophagy, and cancer stem cell-like phenotypes and promoted apoptosis in Huh7 and Mahlavu cells. The sorafenib-resistant clone of Huh7 was effectively inhibited by synergistic combination of both compound in vitro. Eventually, the ovatodiolide→antrocin combination synergistically suppressed the growth of HCC xenografts. Taken together, our findings suggested that ovatodiolide→antrocin combination may represent potential therapeutic approach for patients with advanced HCC.
丝裂原活化蛋白激酶（MAPK）和 PI3K 信号的激活赋予了索拉非尼耐药性，索拉非尼是治疗晚期肝细胞癌（HCC）的主要治疗方法。安托辛和莪术二酮分别是从樟芝和山香圆中分离得到的最有效的次生代谢产物。由于它们在各种实体瘤中可能抑制 MAPK 和 PI3K 信号，这两种天然化合物最近受到了广泛关注。然而，它们的联合应用是否对 HCC 有效仍不清楚。在这里，我们研究了它们单独或联合应用对 HCC 细胞中 MAPK 和 PI3K 信号通路的影响。我们采用一系列体外研究来研究治疗 HCC 的抗癌和干性效果，例如细胞毒性、药物组合指数、迁移、侵袭、集落形成和肿瘤球体形成。采用小鼠异种移植模型评估药物的体内作用。在这项研究中，安托辛和莪术二酮协同抑制了 SNU387、Hep3B、Mahlavu 和 Huh7 细胞系。顺次联合处理 Huh7 和 Mahlavu 细胞先用莪术二酮后用安托辛，比先用安托辛后用莪术二酮、同时给药、单独用安托辛或单独用莪术二酮的效果更强。在 Huh7 和 Mahlavu 细胞系中，莪术二酮→安托辛显著抑制集落形成和增殖，并显著下调 ERK1/2、Akt 和 mTOR 的表达。莪术二酮→安托辛抑制 ERK1/2 和 Akt/mTOR 信号通路，抑制核糖体生物发生、自噬和癌症干细胞样表型，并促进 Huh7 和 Mahlavu 细胞凋亡。体外安托辛和莪术二酮的协同组合有效地抑制了索拉非尼耐药克隆的生长。最终，莪术二酮→安托辛联合抑制 HCC 异种移植物的生长。总之，我们的研究结果表明，莪术二酮→安托辛联合治疗可能为晚期 HCC 患者提供潜在的治疗方法。
