<p>Acute myeloid leukemia (AML) is a highly aggressive hematologic malignancy predominantly seen in adults, with chemotherapy remaining the primary modality for its clinical management. Nevertheless, the efficacy of chemotherapy is often compromised by issues such as drug resistance, toxicity, and long-term adverse effects, particularly in refractory and relapsed patients. Natural products have historically served as a vital source for the development of anti-cancer agents. In this study, we examined the therapeutic potential of pristimerin (PM), a natural bioactive compound derived from the Malvaceae and Portulacaceae families, against acute myeloid leukemia and explored its underlying mechanisms. Our findings indicate that PM effectively suppressed AML cell proliferation, triggered apoptosis, and arrested the cell cycle at the G0/G1 phase. Furthermore, PM was found to enhance erastin-induced ferroptosis via modulation of the ATF3/SLC7A11 axis, characterized by increased iron (Fe²⁺) content, elevated malondialdehyde (MDA) levels, and decreased glutathione (GSH) levels. Mechanistically, PM augmented erastin-induced ferroptosis by regulating the expression of ATF3/SLC7A11, and knockdown of ATF3 significantly attenuated the ferroptosis effect. In conclusion, our results elucidate a previously uncharacterized role of PM in erastin-induced ferroptosis through the modulation of the ATF3/SLC7A11 pathway, indicating its potential as a naturally sourced therapeutic strategy for refractory and relapsed AML.</p> Graphical Abstract <p></p>