姜黄素通过结合 ATOX1 抑制非小细胞肺癌中的铜积累。
Curcumin suppresses copper accumulation in non-small cell lung cancer by binding ATOX1.
机构信息
Pulmonary and Critical Care Medicine, Yantai Shan Hospital, Yantai, 264025, China.
Ministry of Scientific and Technological Innovation, Yantai Hi-tech Industrial Development Zone, Yantai, 264025, China.
出版信息
BMC Pharmacol Toxicol. 2024 Aug 21;25(1):54. doi: 10.1186/s40360-024-00784-0.
BACKGROUND
Non-small cell lung cancer (NSCLC) is associated with intracellular copper accumulation. Antioxidant 1 (ATOX1) is a copper chaperone. This study aimed to analyze the anti-cancer effects of curcumin on the ATOX1-mediated copper pathway in NSCLC.
METHODS
A binding activity between curcumin and ATOX1 was measured using molecular docking. NSCLC cells, A549 and H1299, were treated with different doses of curcumin (10, 20, 40 µM) or DC-AC50 (5, 10, 20 µM) for 24 h. The cell viability and levels of ATOX1, ATP7A and COX17 proteins were observed in cells. Overexpressing ATOX1 in cells was established by pcDNA3.1-ATOX1 transfection for 24 h. The ATOX1 overexpressing cells were treated with 40 µM curcumin or 20 µM DC-AC50 for 24 h to analyze the mechanism of curcumin in NSCLC treatment. Cell viability was measured by CCK-8, and levels of proteins were measured by western blotting. The copper level in cells was labeled by copper sensor-1. Moreover, nude mice models were induced by injection of A549 cells and treated with 20 mg/kg/d DC-AC50 or 40 mg/kg/d curcumin. Tumor growth was observed by measuring tumor volume and tumor weight. The levels of ATOX1, ATP7A and COX17 in tumors were measured by immunohistochemistry and western blotting.
RESULTS
Curcumin bound to ATOX1 (score = -6.1 kcal/mol) and decreased the levels of ATOX1, ATP7A and COX17 proteins in NSCLC cells. The curcumin or DC-AC50 treatment suppressed cell viability by inhibiting the ATOX1-mediated copper signaling in NSCLC cells. The ATOX1 overexpression in cells significantly weakened the effects of curcumin on suppressing copper accumulation and the ATOX1-mediated copper pathway (p < 0.05). In mice models, curcumin or DC-AC50 treatment also suppressed tumor growth by suppressing the ATOX1-mediated copper pathway in tumors.
CONCLUSION
This study demonstrated that curcumin bound ATOX1 to suppress copper accumulation in NSCLC cells, providing a new mechanism of curcumin for NSCLC treatment.
背景
非小细胞肺癌（NSCLC）与细胞内铜积累有关。抗氧化剂 1（ATOX1）是一种铜伴侣。本研究旨在分析姜黄素对 NSCLC 中 ATOX1 介导的铜途径的抗癌作用。
方法
使用分子对接测量姜黄素与 ATOX1 之间的结合活性。用不同剂量的姜黄素（10、20、40 μM）或 DC-AC50（5、10、20 μM）处理 NSCLC 细胞 A549 和 H1299 24 小时。观察细胞中 ATOX1、ATP7A 和 COX17 蛋白的细胞活力和水平。通过 pcDNA3.1-ATOX1 转染 24 小时建立细胞中 ATOX1 的过表达。用 40 μM 姜黄素或 20 μM DC-AC50 处理过表达 ATOX1 的细胞 24 小时，分析姜黄素在 NSCLC 治疗中的作用机制。用 CCK-8 测定细胞活力，用 Western blot 测定蛋白质水平。用铜传感器-1标记细胞内的铜水平。此外，通过注射 A549 细胞诱导裸鼠模型，并以 20mg/kg/d DC-AC50 或 40mg/kg/d 姜黄素进行治疗。通过测量肿瘤体积和肿瘤重量来观察肿瘤生长。用免疫组化和 Western blot 测定肿瘤中 ATOX1、ATP7A 和 COX17 的水平。
结果
姜黄素与 ATOX1 结合（得分=-6.1kcal/mol），降低 NSCLC 细胞中 ATOX1、ATP7A 和 COX17 蛋白水平。姜黄素或 DC-AC50 处理通过抑制 NSCLC 细胞中 ATOX1 介导的铜信号通路抑制细胞活力。细胞中 ATOX1 的过表达显著减弱了姜黄素抑制铜积累和 ATOX1 介导的铜通路的作用（p<0.05）。在小鼠模型中，姜黄素或 DC-AC50 治疗也通过抑制肿瘤中 ATOX1 介导的铜通路抑制肿瘤生长。
结论
本研究表明，姜黄素与 ATOX1 结合抑制 NSCLC 细胞中的铜积累，为姜黄素治疗 NSCLC 提供了新的机制。
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