INTRODUCTION: Pancreatic cancer (PC) has a poor prognosis and limited response to therapies. Combinatorial approaches, such as natural product-based therapies, can enhance anticancer efficacy while minimizing side effects. This study evaluated M. sativa's anticancer properties and its potential as adjunctive therapy with Gemcitabine (GEM) to sensitize PANC-1 cells to chemotherapy. METHODS: The antioxidant activity (AA) and total phenolic content (TPC) of M. sativa extracts (Methanol, Ethyl acetate, and water) were assessed using the DPPH radical scavenging assay. Cytotoxic effects on PANC1 and HUVEC cells were also evaluated by utilizing the MTT assay. Then, apoptosis detection was performed by Annexin V/PI-flow cytometry (FC). Besides, the DNA fragmentation analysis was conducted utilizing agarose gel electrophoresis (AGE). Bcl-2, Bax, and CASP3 expression levels in PANC-1 cells using western blot analysis and qRT-PCR. RESULTS: Herein, DPPH IC50 values for M. sativa extracts (water, MeOH, EtOH) were 76.21, 110.32, and 65.39 μg/ml, respectively. The water extract of M. sativa exhibited the highest TPC (4612.15 ± 119.4 mgGAE/g). The cytotoxicity IC50 values for EtOH M. sativa extract, GEM, and combined GEM with EtOH M. sativa on PANC1 cells were 68.74, 43.53, and 41.22 µg/ml M. sativa + 25 µg/ml GEM, respectively, with no toxicity observed in HUVEC cells. FC analysis revealed that Combining GEM and EtOH M. sativa yielded the highest apoptosis rate (25.6%). Expression changes in Bcl-2, Bax, and CASP3, as well as morphological alterations and DNA fragmentation, indicated apoptotic cell death. CONCLUSION: Our findings suggested that combining M.sativa EtOH extracts with GEM may represent a promising strategy for treating PC.