T-lymphokine-activated killer cell-originated protein kinase (TOPK)是一种丝氨酸/苏氨酸激酶，在胃癌（GC）中高表达并促进肿瘤进展。Paris polyphylla的根茎中分离出的郭氏毒毛茛皂苷七（PPVII）具有抗癌作用。在本研究中，我们探讨了PPVII在GC中的抗肿瘤活性及其机制。通过透射电子显微镜、丙二醛和铁测定实验来检测鞘氨醇引起的铁死亡。通过Western blot和基因变化来检测自噬及其上游信号通路。通过微纳尺度热泳动和药物亲和力响应靶稳定性实验检测PPVII和TOPK的结合。进行小鼠体内模型评估PPVII的治疗作用。PPVII通过诱导自噬介导的铁死亡抑制GC。PPVII促进铁蛋白重链1的降解，而铁蛋白重链1负责介导自噬的铁死亡。PPVII在自噬上游激活Unc-51样自噬激活激酶1（ULK1）。PPVII抑制TOPK的活性，从而减弱下游ULK1的抑制作用。PPVII通过直接结合稳定TOPK非活性二聚体。PPVII抑制肿瘤生长而不引起明显毒性。综上所述，本研究表明PPVII通过靶向TOPK来激活自噬介导的铁死亡，是GC治疗的潜在药物。© 2023约翰威利和子有限公司。

T-lymphokine-activated killer cell-originated protein kinase (TOPK) is a serine-threonine kinase that is overexpressed in gastric cancer (GC) and promotes tumor progression. Polyphyllin VII (PPVII), a pennogenin isolated from the rhizomes of Paris polyphylla, shows anticancer effects. Here, we explored the antitumor activity and mechanism of PPVII in GC. Ferroptosis was detected by transmission electron microscope, malondialdehyde, and iron determination assays. Autophagy and its upstream signaling pathway were detected by Western blot, and gene alterations. The binding of PPVII and TOPK was examined through microscale thermophoresis and drug affinity responsive target stability assays. An in vivo mouse model was performed to evaluate the therapeutic of PPVII. PPVII inhibits GC by inducing autophagy-mediated ferroptosis. PPVII promotes the degradation of ferritin heavy chain 1, which is responsible for autophagy-mediated ferroptosis. PPVII activates the Unc-51-like autophagy-activating kinase 1 (ULK1) upstream of autophagy. PPVII inhibits the activity of TOPK, thereby weakening the inhibition of downstream ULK1. PPVII stabilizes the dimer of the inactive form of TOPK by direct binding. PPVII inhibits tumor growth without causing obvious toxicity in vivo. Collectively, this study suggests that PPVII is a potential agent for the treatment of GC by targeting TOPK to activate autophagy-mediated ferroptosis.© 2023 John Wiley & Sons Ltd.