Modern Research Center for Traditional Chinese Medicine, the Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, China; Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province, Taiyuan, China.
Modern Research Center for Traditional Chinese Medicine, the Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, China; Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Phytomedicine. 2021 Dec;93:153781. doi: 10.1016/j.phymed.2021.153781. Epub 2021 Sep 27.
Hepatocellular carcinoma (HCC) is one of the most extensive and most deadly cancers worldwide. The invasion and metastasis characteristics of HCC dramatically affect the prognosis and survival of HCC patients. Compound Kushen Injection (CKI) is a GMP produced, proverbially applied traditional Chinese medicine formula in China to treat cancer-associated pains, and used as an adjunctive therapy for HCC. Until so far, whether CKI could suppress the metastasis of HCC through regulation of epithelial-mesenchymal transition or metabolic reprogramming is still ambiguous.
In this study, the anti-metastasis effects of CKI were clarified and its pharmacological mechanisms were systematically explored.
Cell invasion and cell adhesion assay were performed in SMMC-7721 cells to assess the anti-metastasis role of CKI, and the histopathological evaluation and biochemical detection were utilized in DEN-induced HCC rats to verify the anti-HCC effect of CKI. Serum and liver samples were analyzed with H NMR metabolomics approach to screen the differential metabolites and further target quantification the content of key metabolites. Finally, western blotting and immunofluorescence assay were applied to verify the crucial signaling pathway involved in metabolites.
CKI markedly repressed the invasion and adhesion in SMMC-7721 cells and significantly improved the liver function of DEN-induced HCC rats. CKI significantly regulated the expression of epithelial-mesenchymal transition (EMT) markers (Vimentin and E-cadherin). Metabolomics results showed that CKI regulated the metabolic reprogramming of HCC by inhibiting the key metabolites (citrate and lactate) and enzymes (HK and PK) in glycolysis process. Importantly, we found that c-Myc mediates the inhibitory effect of CKI on glycolysis. We further demonstrated that CKI inhibits c-Myc expression through modulating Wnt/β-catenin pathway in SMMC-7721 cells and DEN-induced HCC rats. Furthermore, through activating Wnt/β-catenin pathway with LiCl, the inhibitory effects of CKI on HCC were diminished.
Together, this study reveals that CKI intervenes metabolic reprogramming and epithelial-mesenchymal transition of HCC via regulating β-catenin/c-Myc signaling pathway. Our research provides a new understanding of the mechanism of CKI against invasion and metastasis of HCC from the perspective of metabolic reprogramming.
肝细胞癌（HCC）是全球最广泛和最致命的癌症之一。HCC 的侵袭和转移特征严重影响 HCC 患者的预后和生存。复方苦参注射液（CKI）是一种在中国生产的 GMP 产品，是一种传统的中药方剂，用于治疗癌性疼痛，并作为 HCC 的辅助治疗。到目前为止，CKI 是否可以通过调节上皮-间充质转化或代谢重编程来抑制 HCC 的转移仍然不清楚。
本研究旨在阐明 CKI 的抗转移作用，并系统探讨其药理机制。
采用 SMMC-7721 细胞侵袭和细胞黏附实验评价 CKI 的抗转移作用，采用 DEN 诱导的 HCC 大鼠进行组织病理学评价和生化检测验证 CKI 的抗 HCC 作用。采用 H NMR 代谢组学方法分析血清和肝脏样本，筛选差异代谢物，并进一步定量测定关键代谢物的含量。最后，采用 Western blot 和免疫荧光法验证参与代谢物的关键信号通路。
CKI 显著抑制 SMMC-7721 细胞的侵袭和黏附，显著改善 DEN 诱导的 HCC 大鼠的肝功能。CKI 显著调节上皮-间充质转化（EMT）标志物（波形蛋白和 E-钙黏蛋白）的表达。代谢组学结果表明，CKI 通过抑制糖酵解过程中的关键代谢物（柠檬酸和乳酸）和酶（HK 和 PK）来调节 HCC 的代谢重编程。重要的是，我们发现 c-Myc 介导了 CKI 对糖酵解的抑制作用。我们进一步证明，CKI 通过调节 SMMC-7721 细胞和 DEN 诱导的 HCC 大鼠中的 Wnt/β-catenin 通路来抑制 c-Myc 的表达。此外，通过用 LiCl 激活 Wnt/β-catenin 通路，CKI 对 HCC 的抑制作用减弱。
综上所述，本研究揭示了 CKI 通过调节β-catenin/c-Myc 信号通路干预 HCC 的代谢重编程和上皮-间充质转化。我们的研究从代谢重编程的角度为 CKI 抑制 HCC 侵袭和转移提供了新的认识。
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