<p>Pancreatic ductal adenocarcinoma (PDAC) is a cancer with a very poor prognosis, which is bolstered by its resistance and pathogenesis. Quest for PDAC treatments has not left any stones unturned. A well-known naphthoquinone found in many foods, plumbagin (PLGN) demonstrated effectiveness in treating cancer, arthritis, dysmenorrhea, and injuries. We hypothesize therapeutic advantages of PLGN in PDAC and used web resources to assist us identify the targets of PLGN in order to test the idea. After that, in-vitro studies were carried out to comprehend PLGN impacts on two PDAC cell lines. Further, drug combination with PLGN and pre-sensitization by PLGN was also carried out. Findings from network pharmacology investigations revealed PLGN’s targets in PDAC, indicating effect on PDAC survival by targeting crucial signalling. In-vitro tests demonstrated dose-dependent cytotoxicity of PLGN, with IC₅₀ values for Panc-1 and Mia-PaCa-2 cells after 48&#xa0;h were 2.89 ± 0.42µM and 13.10 ± 0.76µM, respectively. Dox-PLGN and Gem-PLGN combination treatment had antagonistic effects, although doxorubicin’s impact was amplified by PLGN pre-sensitization. The EMT transition and genes that cause resistance were the primary pathways impacted by PLGN therapy that enhance Dox effect. Analysis of gene expression and ROS production demonstrated that PLGN had a different effect on both cell lines. Overall, PLGN shows promise as a pre-sensitizing natural substance for PDAC treatment, promoting more research on its effectiveness, tolerability, and possible incorporation into current chemotherapy regimens.</p> Graphical abstract <p></p>