This study investigated the anti-hepatoma molecular mechanism of Alisol G, which is an effective component of the Chinese medicine Alisma orientalis, in the presence of metal ions Cu²⁺ and Fe³⁺ based on c-myc DNA. Here, a combination of Alisol G and metal ions (Cu²⁺, Fe³⁺) to augment anti-hepatoma efficiencies of Alisol G has been identified by methyl thiazolyl tetrazolium (MTT) assay. Network pharmacology revealed that c-myc DNA was the potential target of Alisol G with respect to its anti-hepatoma effects. By performing multi-spectroscopic analyses, we showed that the interaction of Alisol G with c-myc DNA was a process of static quenching. The binding constants and thermodynamic constants indicated that a 1:1 complex was formed between Alisol G and c-myc DNA. Moreover, metal ions strengthened the interaction between Alisol G and c-myc DNA. Molecular docking and molecular dynamics simulation further unveiled that the higher binding affinity between Alisol G-Fe³⁺ complex and c-myc DNA as compared to Alisol G-Cu²⁺ complex. This probably resulted from the polarization of metal ions and the structural flexion of Alisol G. The C22-O31-H76 and C18-O32-H77 of Alisol G were key groups in the interaction with c-myc DNA. Addition of metal ion, had greatly changed the c-myc DNA-binding domain of Alisol G while didn’t affect the kinetic stability of the interaction, thus facilitating the insertion of Alisol G into c-myc DNA A-T base pair. Importantly, the DG113 of c-myc DNA was important for its binding to metal ions. Together, our findings suggested that Alisol G in combination with metal ions may be an efficient and promising option for the treatment of liver cancer.

本研究基于c-myc DNA ，在金属离子Cu ²⁺和Fe ³⁺存在下，研究了中药泽泻的有效成分Alisol G的抗肝癌分子机制。这里，Alisol G 和金属离子（Cu ²⁺、Fe ³⁺) 已通过甲基噻唑基四唑 (MTT) 测定确定了 Alisol G 的抗肝癌效率。网络药理学表明 c-myc DNA 是 Alisol G 抗肝癌作用的潜在靶点。通过进行多光谱分析，我们表明 Alisol G 与 c-myc DNA 的相互作用是一个静态淬灭过程。结合常数和热力学常数表明 Alisol G 和 c-myc DNA 之间形成了 1:1 的复合物。此外，金属离子加强了 Alisol G 和 c-myc DNA 之间的相互作用。分子对接和分子动力学模拟进一步表明，与 Alisol G-Cu ²⁺相比，Alisol G- Fe ³⁺复合物和 c-myc DNA之间的结合亲和力更高复杂的。这可能是由于金属离子的极化和 Alisol G 的结构弯曲所致。 Alisol G 的 C22-O31-H76 和 C18-O32-H77 是与 c-myc DNA 相互作用的关键基团。添加金属离子，极大地改变了Alisol G的c-myc DNA结合域，同时不影响相互作用的动力学稳定性，从而促进了Alisol G插入c-myc DNA AT碱基对。重要的是，c-myc DNA 的 DG113 对其与金属离子的结合很重要。总之，我们的研究结果表明 Alisol G 与金属离子结合可能是治疗肝癌的有效且有前景的选择。