Proteomics, Transcriptomics, and Phosphoproteomics Reveal the Mechanism of Talaroconvolutin-A Suppressing Bladder Cancer via Blocking Cell Cycle and Triggering Ferroptosis

Talaroconvolutin-A (TalaA) is a compound from the endophytic fungus T. convolutispora of the Chinese herbal medicine Panax notoginseng. Whether TalaA exerts Anticancer activity in bladder Cancer remains unknown. We explored Anticancer function and studied the pharmacological mechanism of TalaA from transcriptomic, proteomic, phosphoproteomic, and molecular interaction perspectives. Using CCK8 assay, EdU staining, crystal violet staining, flow cytometry, living/dead cell staining, and western blotting we studied the Anticancer activity of TalaA in vitro. For in vivo analysis, we performed xenograft tumor implantation. The anti-tumor effects of TalaA were evaluated through hematoxylin and eosin (H&E) and immunohistochemistry staining and pathological analysis. In high-throughput omics detection, proteomics was conducted to detect changes in the protein profile; transcriptomics was performed used to detect changes in mRNA abundance; phosphoproteomics was used to detect changes in protein phosphorylation. We found TalaA inhibited tumor cell proliferation, DNA replication, and colony formation in a dose-dependent manner in bladder Cancer cells. The IC₅₀ values of TalaA on SW780 and UM-UC-3 cells were 5.7 and 8.2 μM respectively. TalaA (6.0 mg/kg) significantly repressed the growth of xenografted tumors and did not affect the body weight nor cause obvious hepatorenal toxicity. In pharmacological mechanism, TalaA arrested the cell cycle by downregulating cyclinA2, cyclinB1, and AURKB and upregulating p21/CIP. TalaA also elevated intracellular Reactive Oxygen Species and upregulated transferrin (TF) and heme oxygenase 1 (HMOX1) to induce Ferroptosis. Moreover, TalaA was able to bind to MAPKs (MAPK1, MAPK8, and MAPK14) to inhibit the phosphorylation of *SP* motif sites of transcription regulators. This study revealed that TalaA inhibited bladder Cancer mainly by arresting cell cycle to suppress proliferation and triggering Ferroptosis to cause cell death. Conclusively, TalaA would be a potential candidate for treating bladder Cancer by targeting MAPKs, suppressing the cell cycle, and inducing Ferroptosis.

Bladder cancer; Pharmacology; Phosphoproteomics; Proteomics; Transcriptomics.