Gypenoside LI Inhibits Renal Cancer Proliferation by Affecting LipidMetabolism through the Upregulation of Dual-Specificity Phosphatase 1

Introduction: Gynostemma pentaphyllum is commonly used in southern China to treat hyperlipidemia and tumors, primarily due to its active saponin compounds. This study aimed to analyze the inhibitory effect of gypenoside LI (Gyp LI) on clear cell renal cell carcinoma (ccRCC), which is mediated through DUSP1-related lipid metabolism. Materials and Methods: The concentration of Gyp LI required to inhibit the ccRCC cell lines 769-P and ACHN was determined using a CCK8 assay, and DUSP1 upregulation was confirmed by transcriptomic sequencing, RT-qPCR, and Western blot analysis. The roles of DUSP1 and Gyp LI in regulating cell proliferation were further validated using CCK-8, colony formation, and flow cytometry assays. . Additionally, we established a nude mouse xenograft tumor model using ACHN cells that overexpress DUSP1. We assessed the effects of Gyp LI and DUSP1 on tumor growth through histopathological examinations. We employed untargeted tissue metabolomics to identify metabolic pathways influenced by Gyp LI via upregulation of DUSP1. Results: CCK-8 experiments showed that Gyp LI inhibited the proliferation of ccRCC cells. Additionally, transcriptome sequencing revealed that DUSP1 expression increased following Gyp LI intervention. We transfected cells with siDUSP1 and ovDUSP1 and assessed cell proliferation using CCK8, colony formation, flow cytometry, and WB analyses of apoptosis-related proteins, confirming that both ovDUSP1 and Gyp LI inhibited ccRCC cell proliferation. Animal studies revealed that both ovDUSP1 and Gyp LI inhibited tumor growth. Non-targeted metabolomics analysis revealed that Gyp LI affected seven metabolites associated with ccRCC treatment by upregulating DUSP1. Conclusion: Gyp LI upregulates DUSP1 to suppress the metabolism of choline, linoleic acid, and alpha-linolenic acid, ultimately suppressing the incidence and progression of ccRCC.