BACKGROUND AND AIM: Platycodin D (PD), a major bioactive saponin isolated from the traditional Chinese medicine (TCM) Platycodon grandiflorus, has shown promising therapeutic potential against non-small cell lung cancer (NSCLC). However, the functional mechanisms of PD in NSCLC progression remains unclear. This study aimed to explore the pharmacological mechanism of PD against NSCLC. MATERIALS AND METHODS: Thermal proteome profiling (TPP) approach, molecular docking, cellular thermal shift assay (CETSA) and peptide-centric local stability assay (PELSA) were employed to identify the potential binding target of PD. Subsequent Western Blot and immunoprecipitation-Western Blot (IP-WB) experiments were conducted to investigate the downstream signaling pathways of the target. Furthermore, proteomic and ubiquitinomic profiling of PD-treated cells were performed to investigate its functions on global. RESULTS: Replication factor C subunit 4 (RFC4) was identified as a potential binding target of PD by TPP and their binding sites were further exposed by PELSA. PD-RFC4 complex promotes the degradation of Notch1 and Notch3 by reducing nuclear entry of their domains. Compared with control treatment, the differentially expressed proteins induced by PD were found to be primarily involved in ferroptosis, ubiquitination, platinum drug resistance and ribosome-related processes. The ubiquitin proteome analysis revealed that proteins associated with the Notch pathway underwent ubiquitin modifications. CONCLUSIONS: PD binds to RFC4 and inhibits its activity, leading to downregulation of the Notch signaling pathway, ultimately triggering cancer cell apoptosis. PD is a natural product with potential therapeutic value for NSCLC.
Zhang et al. (Fri,) studied this question.