胰腺癌作为一种恶性消化系统肿瘤，具有较高的死亡率。黄体芽菌素是从传统中草药蒿玄参（Xanthium strumarium L）中提取的一种倍萜内酯单体化合物。已有报道显示黄体芽菌素对视网膜母细胞瘤、胶质瘤、肝癌、结肠癌、肺癌和乳腺癌等多种癌细胞表现出抑制作用。然而，在胰腺癌细胞中，目前仅有一份关于黄体芽菌素对MIA PaCa-2细胞中前列腺素E2合成抑制和caspase 3/7活化诱导的报告存在，需要进一步的系统的体外和体内研究及相关机制的探索。本研究旨在探索黄体芽菌素在胰腺癌细胞中的体内外效果及其分子机制。通过细胞计数试剂盒-8（CCK-8）实验、乳酸脱氢酶（LDH）实验、二酯类荧光素乙酸缀合酯（CFDA SE）细胞增殖实验、克隆形成实验、伤口愈合实验、Transwell实验、Annexin V-FITC/propidium iodide（PI）双染实验、Hoechst核染色、Western blot分析、磷酸化蛋白质组学和活性氧（ROS）测量，评估了黄体芽菌素在胰腺癌细胞中的抗癌效果和机制。使用裸小鼠模型研究了黄体芽菌素在胰腺癌细胞中的体内抗癌效果。本研究表明，黄体芽菌素能够阻止胰腺癌细胞的增殖和转移，并触发磷脂酰丝氨酸（PS）的外露、染色质浓缩和caspase的活化，从而诱导细胞凋亡。磷酸化蛋白质组学分析表明，黄体芽菌素抑制了与增殖相关的蛋白质RBL1的磷酸化，而氧化应激能导致RBL1的磷酸化去除。进一步的研究发现，黄体芽菌素显著上调了胰腺癌细胞中的ROS水平，抗氧化剂N-乙酰半胱氨酸（NAC）可以逆转黄体芽菌素诱导的细胞增殖抑制和凋亡。此外，黄体芽菌素在裸小鼠的异种移植模型中可以抑制胰腺癌细胞的生长，对小鼠毒性较低。黄体芽菌素可能通过ROS/RBL1信号通路抑制胰腺癌细胞的增殖并触发凋亡。版权 © 2023 Elsevier GmbH。保留所有权利。

As a malignant digestive system tumor, pancreatic cancer has a high mortality rate. Xanthatin is a sesquiterpene lactone monomer compound purified from the traditional Chinese herb Xanthium strumarium L. It has been reported that Xanthatin exhibits inhibitory effects on various cancer cells in retinoblastoma, glioma, hepatoma, colon cancer, lung cancer, as well as breast cancer. However, in pancreatic cancer cells, only one report exists on the suppression of Prostaglandin E2 synthesis and the induction of caspase 3/7 activation in Xanthatin-treated MIA PaCa-2 cells, while systematic in vitro and in vivo investigations and related mechanisms have yet to be explored.This research aims to explore the in vitro and in vivo effects of Xanthatin on pancreatic cancer and its molecular mechanisms.The anticancer effects and mechanisms of Xanthatin on pancreatic cancer cells were assessed through employing cell counting kit-8 (CCK-8) assay, lactate dehydrogenase (LDH) assay, carboxyfluorescein diacetate succinimidyl ester (CFDA SE) cell proliferation assay, colony formation assay, wound healing assay, transwell assay, Annexin V-FITC/propidium iodide (PI) dual staining, Hoechst nuclear staining, Western blot analysis, phosphoproteomics, and reactive oxygen species (ROS) measurement. The in vivo anticancer effects of Xanthatin on pancreatic cancer cells were studied using a nude mouse model.The present study showed that Xanthatin can prevent the proliferation and metastasis of pancreatic cancer cells and trigger the exposure of phosphatidylserine (PS), chromatin condensation, and caspase activation, thereby inducing apoptosis. Phosphoproteomic analysis indicated that Xanthatin inhibits the phosphorylation of the proliferation-associated protein RBL1, and oxidative stress can lead to RBL1 dephosphorylation. Further investigation revealed that Xanthatin significantly upregulates ROS levels in pancreatic cancer cells, and the antioxidant N-acetylcysteine (NAC) can reverse Xanthatin-induced cell proliferation inhibition and apoptosis. In addition, Xanthatin can suppress pancreatic cancer cell growth in a xenograft nude mouse model with low toxicity to the mice.Xanthatin may inhibit the proliferation of pancreatic cancer cells and trigger apoptosis through the ROS/RBL1 signaling pathway.Copyright © 2023 Elsevier GmbH. All rights reserved.