Fu Zheng Xiao Yu San Jie Decoction affects the proliferation of renal cell carcinoma via regulating E2F5 gene.

BACKGROUND: Renal cell carcinoma (RCC) is a prevalent disease within the urinary system, characterized by high mortality rates. Its incidence is increasing each year, posing a significant threat to public health. Currently, the causes of RCC are not fully understood, and there are many limitations in treatment options. Recently, traditional Chinese medicine has gained considerable attention in cancer treatment. Fu Zheng Xiao Yu San Jie Decoction (FZXYSJD) is a well-established treatment for RCC, used for many years at The First Affiliated Hospital of Guangzhou University of Chinese Medicine, and is recognized for its efficacy by both physicians and patients. This study aims to employ methods such as network pharmacology and cell experiments to investigate how FZXYSJD works in treating RCC. Additionally, it seeks to identify new targets that may influence the disease's progression. METHODS: First, FZXYSJD drug serum was prepared for cell experiments, including the Cell Counting Kit-8 (CCK-8) assay and colony formation assays. Second, Gene Expression Profiling Interactive Analysis (GEPIA) was applied to explore the specific mechanism of FZXYSJD's suppressive effect in RCC. Western blot analyzed the expression level of E2F transcription factor 5 (E2F5) after treatment with FZXYSJD drug serum in RCC cells. Third, the effect of small interfering RNA targeting E2F5 (siE2F5) on RCC cell proliferation was verified using CCK-8 and colony formation assays. Fourth, the herb identification components of FZXYSJD were retrieved from the Chinese Pharmacopoeia and those components of FZXYSJD that were decocted by high-performance liquid chromatography-mass spectrometry (LC-MS) were detected. Finally, the binding potential of the active ingredient in FZXYSJD to E2F5 was confirmed through molecular docking. RESULTS: FZXYSJD was found to suppress the proliferation of renal clear cell carcinoma cell lines (786O, ACHN) and promote the E2F5 expression. In addition, our findings indicated that low levels of E2F5 expression promoted the proliferation of RCC. Finally, through LC-MS analysis, we found that the components that could be detected after FZXYSJD water decoction were calycosin-7-O-β-D-glucoside, rosmarinic acid, ginsenoside Rg1, liquiritin, glycyrrhizic acid, emodin and polydatin. And the binding potential between those components in FZXYSJD and E2F5 was confirmed through molecular docking. CONCLUSIONS: FZXYSJD affects cell proliferation in RCC through the E2F5 gene.