Rhizoma paridis is a traditional Chinese medicine that has been reported to have anti-cancer activity. However, the antitumor effect of pennogenin 3-O-f3-chacotrioside (Formular: C45H72O17), a steroidal saponin isolated from its extract, in lung cancer have not been well studied, its potential mechanisms remain unclear. In this paper, its effect on the cell viability of non-small cell lung can-cer (NSCLC) was examined using the Cell Counting Kit-8 assay, the potential target proteins were screened by antibody microarrays, the potential pharmacological mechanisms were predicted using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein IP: 203.8.109.20 On: Thu, 20 Oct 2022 11:01:36 interaction (PPI) analyses.Copyright:Moreover, tofurAmericanther validateScientificthe findings,Publisherscell apoptosis and cell cycle arrest were measured by flow cytometry, cllinvasioand migration were measured by Transwell Delivered by In genta assays, and apoptosis-related protein expression levels were measured using western blotting. The results showed that, after treatment by pennogenin 3-O-f3-chacotrioside, the cell viability was significantly reduced. In addition, thirteen target proteins related to apoptosis and platinum drug resistance were screened out, and their PPIs were constructed. Moreover, it was confirmed to have effects in inducing apoptosis and promoting cell cycle arrest in NSCLC cells. The expression lev-els of apoptosis-related proteins, such as caspase 8, caspase 9, phospho-ERK1/2, phospho-p38, phospho-Akt, were decreased following the treatment, whereas the expression levels of cleaved caspase 3 and cytochrome C were increased. The findings of the present study highlight the poten-tial of pennogenin 3-O-f3-chacotrioside as a novel therapeutic agent.