Integrated multi-omics analysis of Periplocymarin to identify the mechanism of p38α MAPK phosphorylation inhibition in Non-small cell lung cancer

Background: Periplocymarin (PPM) is a plant-derived natural product, which is isolated and purified from the dried root bark of Periploca sepium. Previous studies have shown that Periplocymarin exhibits significantly and broad-spectrum anti-tumor pharmacological activity. To investigate the anti-tumor activity and mechanism of the natural product Periplocymarin against Non-small cell lung cancer (NSCLC). Methods: This study screened a library of 1459 anti-tumor natural products using CCK-8 assays in three NSCLC cell lines and two normal lung epithelial cell lines, identifying Periplocymarin as a selective candidate against NSCLC. The effects of Periplocymarin on NSCLC proliferation, apoptosis, and migration were assessed in cell lines and patient-derived organoid (PDO) models. Transcriptome sequencing, network pharmacology, proteome thermal stability profiling (TPP), Microscale Thermophoresis (MST) and Molecular Simulation were integrated to identify the target of Periplocymarin. Western blot and CCK-8 assays were performed to verify Periplocymarin’s effect on p38α mitogen-activated protein kinase (p38α MAPK, MAPK14) phosphorylation. In vivo anti-tumor efficacy and safety of Periplocymarin were evaluated in nude mouse xenograft and patient-derived xenograft (PDX) models. Results: Periplocymarin exhibited concentration-dependent inhibition of proliferation, promoted apoptosis, and reduced cell migration, while also suppressing growth in organoids. Integrated analyses indicated MAPK14 as the target, and Western blot confirmed markedly inhibition of MAPK14 phosphorylation by Periplocymarin. MAPK14 knockdown attenuated Periplocymarin’s anti-tumor effects. In vivo , Periplocymarin significantly inhibited tumor growth, with decreased Ki-67 and phospho-MAPK14 (p-MAPK14) expression in tumor tissues and no evident organ toxicity or hematological abnormalities. Conclusions: Periplocymarin, by predominantly inhibiting MAPK14 phosphorylation, presents a robust anti-tumor effect in vitro and in vivo . These findings provide a theoretical and experimental basis for the application of Periplocymarin and for the development of novel anticancer drugs targeting MAPK14 in NSCLC.