School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Jiangsu Key Laboratory of Pediatric Respiratory Disease, Institute of Pediatrics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Med Oncol. 2022 Sep 29;39(12):221. doi: 10.1007/s12032-022-01797-7.
Sorafenib (SOR) is currently the first line molecular targeting agent for advanced liver cancer therapy. Unfortunately, the insensitivity of liver cancer patients to SOR relatively limits its effectiveness. Huaier (HUA), a natural medicinal parasitic fungus found on the Sophora japonica Linn., has been widely employed as an adjuvant medication for numerous malignancies due to its potent anti-tumoral properties. This study aims to elucidate the enhancing therapeutic efficacy of HUA on SOR treatment in hepatocellular carcinoma (HCC) cells and mouse models. The CCK-8, clone formation, flow cytometry, immunofluorescence, transmission electron microscopy, western blot, bioinformatic analysis, and xenograft tumor assays were performed to evaluate the synergistic anti-hepatoma efficacy and mechanisms of HUA-SOR combination treatment on HCC cells. The results revealed combination treatment further inhibited proliferation, promoted apoptosis, enhanced autophagy of HCC cells, and suppressed the growth of transplanted tumors in mice, compared with either HUA or SOR treatment alone. For Hep3B and Huh7 cells, the optimal synergistic doses of HUA in combination with SOR were 8 mg/mL + 4 μM and 4 mg/mL + 2 μM, with combination index values of 0.646 and 0.588, respectively. Additionally, the underlying mechanisms might be related to biological processes that are mediated by mammalian target of rapamycin (mTOR). The combination treatment downregulated the protein expression levels of p-mTOR, p-p70S6K, p62, and upregulated the protein expression levels of Beclin-1 and LC3B-II. The mTOR activator MHY1485 attenuated the effect of HUA-SOR combination by inhibiting autophagy, suggesting HUA may potentiate the sensitivity of HCC cells to SOR by partially inducing mTOR-mediated autophagic cell death. These findings might provide a rationale experimental foundation for clinical applications of HUA with SOR.
索拉非尼（SOR）是目前晚期肝癌治疗的一线分子靶向药物。不幸的是，肝癌患者对 SOR 的不敏感相对限制了其疗效。槐耳（HUA），一种天然药用寄生真菌，发现于槐属植物，由于其强大的抗肿瘤特性，已被广泛用作多种恶性肿瘤的辅助药物。本研究旨在阐明 HUA 增强 SOR 治疗肝癌细胞及小鼠模型的治疗效果。通过 CCK-8、克隆形成、流式细胞术、免疫荧光、透射电子显微镜、western blot、生物信息学分析和异种移植肿瘤检测，评估 HUA-SOR 联合治疗对肝癌细胞的协同抗肝癌疗效及机制。结果表明，与单独使用 HUA 或 SOR 相比，联合治疗进一步抑制了肝癌细胞的增殖，促进了肝癌细胞的凋亡，增强了自噬，并抑制了小鼠移植瘤的生长。对于 Hep3B 和 Huh7 细胞，HUA 与 SOR 联合的最佳协同剂量分别为 8 mg/mL+4 μM 和 4 mg/mL+2 μM，组合指数值分别为 0.646 和 0.588。此外，潜在的机制可能与雷帕霉素（mTOR）介导的生物学过程有关。联合治疗下调了 p-mTOR、p-p70S6K、p62 的蛋白表达水平，上调了 Beclin-1 和 LC3B-II 的蛋白表达水平。mTOR 激活剂 MHY1485 通过抑制自噬减弱了 HUA-SOR 联合治疗的作用，表明 HUA 可能通过部分诱导 mTOR 介导的自噬细胞死亡来增强 HCC 细胞对 SOR 的敏感性。这些发现可能为 HUA 与 SOR 的临床应用提供实验依据。
