Study on Anti-Melanoma Mechanism of Sparganii Rhizoma Extract Inhibiting Glycolysis Rate-limiting Enzyme

  OBJECTIVE  To explore the molecular mechanism of Sparganii Rhizome anti-melanoma and its effective material basis.

  METHODS  The active ingredients and corresponding targets of Sparganii Rhizome were retrieved through the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), and the melanoma disease targets were retrieved by GeneCards database. The drug-disease target protein-protein interaction (PPI) was constructed by the cytoscape 3.7.2 software. The core targets were screened out and uploaded to the DAVID platform for KEGG pathway enrichment analysis. Further verification was carried out through cell proliferation and activity detection (CCK-8) experiment, EdU proliferation testing experiment, Seahorse XF cell mito stress analysis experiment, metabolites and enzyme activity detection kit experiment and Western blot.

  RESULTS  Obtained from the TCMSP database, the five active ingredients in Sparganii Rhizome, corresponding to a total of 70 targets, constructed a PPI network with disease targets and further screened to obtain a key network composed of 86 core targets. The results of enrichment analysis of the KEGG signaling pathway showed that Sparganii Rhizome treatment of melanoma mainly involved metabolic-related pathways, p53 signal pathways, AMPK signal pathways, MAPK signal pathways, PI3K/Akt signal pathways, apoptosis signal pathways, etc. The results of in vitro experiments showed that the Sparganii Rhizome extract (SLTQW) inhibits the proliferation of melanoma A375 cells in a dose-depentent manner and reduces the rate of extracellular acidification of cells and the production of glycolytic metabolites pyruvate, lactic acid, ATP. The results of the kit and Western blot showed that the SLTQW inhibited the activity of PK and reduced the protein expression levels of PKM2, LDHA, MCT4.

  CONCLUSION  SLTQW has obvious anti-melanoma effect, and its mechanism may be related to inhibiting the glycolysis-related proteins PKM2, LDHA, MCT4 expression and interfering with tumor cell glycolysis metabolism.