Helmueller Sarah, Song Xinxin, Kim Dong-Hyun, Lee Yong J
Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
Department of Surgery, UT Southwestern Medical Center, Dallas, TX, USA.
Int J Hyperthermia. 2025 Dec;42(1):2544017. doi: 10.1080/02656736.2025.2544017. Epub 2025 Aug 11.
The combination of ferroptotic agent artesunate (ART) and apoptotic agent rhTRAIL (recombinant human tumor necrosis factor-related apoptosis-inducing ligand) has been shown to synergistically enhance apoptosis in various cancer cell lines via crosstalk between the endoplasmic reticulum (ER) stress response, the rhTRAIL-induced extrinsic cell death receptor pathway, and the intrinsic BID-Bax-mitochondrial-dependent-apoptosis pathway. This synergistic interaction has been demonstrated to be effective in multiple types of cancer cell lines, making artesunate combined with rhTRAIL a promising second-line therapy for colon cancer patients after cytoreductive surgery and chemotherapeutic treatments. To further enhance the second-line therapy's tumoricidal effect, a multimodal therapy was developed by combining artesunate, rhTRAIL, and hyperthermic conditions where samples were treated at 42 °C for 1 h.
The effects of this therapy were tested in human colon carcinoma HCT116 and pancreatic adenocarcinoma BxPC-3 cell models. The cytotoxic and synergistic effects were analyzed using fluorescence microscopy, cell survival assays, and protein analysis through Western blotting.
Our findings demonstrated a significant enhancement of apoptosis when artesunate and rhTRAIL treatments were combined with heat exposure. The synergistic and apoptotic effect of the agents was effectively abrogated in BID-deficient and BID mutant-type cells as well as Bax-deficient cells, but not Bak-deficient cells.
The results suggest that BID acts as a key gatekeeper molecule of apoptosis during hyperthermia-based multimodal treatment. These findings raise important questions about the underlying mechanisms of heat-induced apoptosis and its involvement in orchestrating various cellular stress pathways.
已证明铁死亡诱导剂青蒿琥酯（ART）与凋亡诱导剂重组人肿瘤坏死因子相关凋亡诱导配体（rhTRAIL）联合使用，可通过内质网（ER）应激反应、rhTRAIL诱导的外源性细胞死亡受体途径和内源性BID-Bax-线粒体依赖性凋亡途径之间的相互作用，协同增强多种癌细胞系中的细胞凋亡。这种协同相互作用已在多种癌细胞系中得到证实，使青蒿琥酯联合rhTRAIL成为结肠癌患者在减瘤手术和化疗后的一种有前景的二线治疗方法。为了进一步增强二线治疗的杀瘤效果，通过将青蒿琥酯、rhTRAIL与热疗条件相结合开发了一种多模式疗法，其中样品在42°C下处理1小时。
在人结肠癌HCT116和胰腺腺癌BxPC-3细胞模型中测试了该疗法的效果。使用荧光显微镜、细胞存活分析和蛋白质印迹法进行蛋白质分析来分析细胞毒性和协同作用。
我们的研究结果表明，青蒿琥酯和rhTRAIL处理与热暴露相结合时，细胞凋亡显著增强。在BID缺陷型和BID突变型细胞以及Bax缺陷型细胞中，这些药物的协同和凋亡作用被有效消除，但在Bak缺陷型细胞中未被消除。
结果表明，在基于热疗的多模式治疗期间，BID作为细胞凋亡的关键守门分子。这些发现引发了关于热诱导细胞凋亡的潜在机制及其在协调各种细胞应激途径中的作用的重要问题。
