Wu Quhui, Li Da, Sun Taoli, Liu Jian, Ou Huiping, Zheng Lei, Hou Xuyang, Li Wenqun, Fan Fuyuan
Department of Respiratory Medicine, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, China.
Medical School, Hunan University of Chinese Medicine, Changsha, 410208, P. R. China.
J Cancer. 2021 Sep 9;12(21):6576-6587. doi: 10.7150/jca.62779. eCollection 2021.
Bai-He-Gu-Jin-Tang (BHGJT) is a classic Chinese formula used to treat lung cancer, while the underlying molecular mechanism remains obscure. The aim of the study was to investigate the molecular mechanism of BHGJT on lung cancer and demonstrate the potential for synergistic treatment combining BHGJT with conventional therapy. Cell viability assay, colony formation assay and EdU assay were used to determine the effects of BHGJT, and a subcutaneous xenograft model was used to evaluate the effect. Cell cycle analysis, apoptosis rate analysis, immunohistochemical and immunofluorescent staining, Western blot assays and network pharmacology-based analysis were used to explore the underlying mechanisms. We found that BHGJT inhibited cell proliferation via a dose-dependent pathway and obviously hindered tumor growth in lung cancer. Cell cycle arrest and apoptosis were pronouncedly induced by BHGJT via dysregulation of the cell cycle regulators CDK4 and Cyclin D1 and dysregulation of apoptosis-associated proteins, such as cleaved caspase 3/9 and the BCL-2 family. Based on a network pharmacology-based analysis and experimental evidence, we demonstrated that the AKT/GSK3β/β-catenin signaling pathways were responsible for BHGJT-induced apoptosis in lung cancer cells. Additionally, autophagy was induced by BHGJT via the AMPK/mTORC1/ULK1 signaling pathway, and blocking autophagy with either chloroquine or a ULK1 inhibitor increased the killing efficiency of BHGJT in lung cancer cells. Our findings indicate that the BHGJT formula efficiently inhibits lung cancer growth and represents a potential complementary and alternative treatment for lung cancer.
百合固金汤（BHGJT）是一种用于治疗肺癌的经典中药方剂，但其潜在的分子机制仍不清楚。本研究的目的是探讨百合固金汤对肺癌的分子机制，并证明百合固金汤与传统疗法联合协同治疗的潜力。采用细胞活力测定、集落形成测定和EdU测定来确定百合固金汤的作用，并使用皮下异种移植模型来评估其效果。通过细胞周期分析、凋亡率分析、免疫组织化学和免疫荧光染色、蛋白质免疫印迹分析以及基于网络药理学的分析来探索潜在机制。我们发现百合固金汤通过剂量依赖性途径抑制细胞增殖，并明显抑制肺癌肿瘤生长。百合固金汤通过细胞周期调节因子CDK4和细胞周期蛋白D1的失调以及凋亡相关蛋白（如裂解的半胱天冬酶3/9和BCL-2家族）的失调，显著诱导细胞周期停滞和凋亡。基于网络药理学分析和实验证据，我们证明AKT/GSK3β/β-连环蛋白信号通路是百合固金汤诱导肺癌细胞凋亡的原因。此外，百合固金汤通过AMPK/mTORC1/ULK1信号通路诱导自噬，用氯喹或ULK1抑制剂阻断自噬可提高百合固金汤对肺癌细胞的杀伤效率。我们的研究结果表明，百合固金汤配方可有效抑制肺癌生长，是一种潜在的肺癌互补和替代治疗方法。
