硫唑嘌呤的新型再利用用于肾上腺皮质癌的治疗，可能通过SLC7A11/xCT-hsa-miR-92a-3p-OIP5-AS1网络途径实现

Novel repurposing of sulfasalazine for the treatment of adrenocortical carcinomas, probably through the SLC7A11/xCT-hsa-miR-92a-3p-OIP5-AS1 network pathway

SURGERY

影响因子:2.7

分区:医学2区 / 外科2区

发表日期:2025 Jan

作者: Chitra Subramanian, Kelli McNamara, Seth W Croslow, Yanqi Tan, Daniel Hess, Katja Kiseljak-Vassiliades, Margaret E Wierman, Jonathan V Sweedler, Mark S Cohen

DOI: 10.1016/j.surg.2024.07.075

摘要

近年来对肾上腺皮质癌（ACC）的多基因组分析发现，SLC7A11/xCT是一种新型生物标志物。美国食品药品监督管理局（FDA）批准的抗炎药硫唑嘌呤（SAS）通过阻断SLC7A11表达诱导铁死亡（ferroptosis）。我们假设SAS可以被重新用于靶向ACC细胞。利用Gene Expression Profiling Interactive Analysis（GEPIA）分析SLC7A11的表达及其与ACC生存的关联。采用验证的ACC细胞系NCI-H295R、ACC1和ACC2，在二维培养中进行研究。包括CellTiter-Glo细胞活性检测、Western blot分析（WB）检测凋亡及其他靶蛋白变化、逆转录聚合酶链反应检测甾体生成酶变化、C11BODIPY检测脂质过氧化以及质谱分析脂质变化。GEPIA数据库分析显示，SLC7A11及其相关长链非编码RNA OAP5-AS1在ACC肿瘤中高表达（n=77对128，P<.05），与较差的总生存期和无病生存期相关，风险比分别为4.3和5.2（SLC7A11），4.8和2.7（OAP5-AS1）。SAS处理72小时后，ACC细胞系的半数抑制最大浓度（IC50）范围为412 nM（ACC1）至799 nM（ACC2），均表现出多聚ADP-核糖聚合酶（PARP）裂解、p-Akt和p-ERK上调、GPX4和SLC7A11下调（P<.05，WB分析）。球体形成、迁移和侵袭实验显示抑制作用，脂质过氧化（C11BODIPY）、细胞内铁增加、氧化应激诱导及氧化多不饱和脂肪酸磷脂的显著上调（P<.05，质谱分析）提示铁死亡的诱导。SAS下调ACC中的tSLC7A11，靶向Akt/ERK通路和脂质代谢，诱导细胞死亡，提示其具有潜在的转化医学应用价值，值得进一步研究以明确其在ACC中的治疗潜力。

Abstract

Recent multigenomic analysis of adrenocortical carcinomas (ACCs) identified SLC7A11/xCT as a novel biomarker. The Food and Drug Administration-approved anti-inflammatory drug, sulfasalazine (SAS), induces ferroptosis by blocking SLC7A11 expression. We hypothesize that SAS could be repurposed to target ACC cells.Expression of SLC7A11 and its association with ACC survival was analyzed using Gene Expression Profiling Interactive Analysis (GEPIA). The validated ACC cell lines NCI-H295R, ACC1, and ACC2 were grown in 2D culture. In vitro studies included the CellTiter-Glo assay to calculate viability, Western blot (WB) analysis for apoptosis and other target protein changes, reverse transcriptase polymerase chain reaction for steroidogenic enzyme changes, C11BODIPY for lipid peroxidation, and mass spectrometry for changes in lipids.The Cancer Genome Atlas Program database analysis in GEPIA showed that SLC7A11 and linked long noncoding RNA OAP5-AS1 are highly expressed in ACC tumors versus normal adrenals (n = 77 vs 128; P < .05). This was associated with poor overall and disease-free survival with hazard ratios of 4.3 and 5.2 for SLC7A11 and 4.8 and 2.7 for OAP5-AS1, respectively. ACC cell line half-inhibitory maximum concentration values after 72-hour SAS treatment ranged from 412 nM (ACC1) to 799 nM (ACC2), and all showed cleavage of poly (ADP-ribose) polymerase, upregulation of p-Akt and p-ERK, and downregulation of GPX4 and SLC7A11 (P < .05) by WB analysis. Sphere formation, migration, and invasion assay showed inhibition, and lipid peroxidation using C11BODIPY, increase in intracellular iron, induction of oxidative stress, and significant upregulation of oxidized polyunsaturated fatty acid phospholipids (P < .05 each) by mass spectrometry suggests induction of ferroptosis.SAS downregulates tSLC7A11 in ACCs, targets the Akt/ERK pathway and lipid metabolism, and induces cell death in vitro, warranting additional translational studies to define its therapeutic potential in ACC.