Kim Jin Sun, Choi Won-Mook, Kim Ha-Il, Chung Sung Won, Choi Jonggi, Lee Danbi, Kim Kang Mo
Department of Gastroenterology, Liver Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
J Liver Cancer. 2025 Mar;25(1):79-90. doi: 10.17998/jlc.2024.12.27. Epub 2025 Jan 14.
BACKGROUNDS/AIMS: Hepatocellular carcinoma (HCC) is a malignant cancer with an increasing incidence worldwide. Although numerous efforts have been made to identify effective therapies for HCC, current strategies have limitations. We present a new approach for targeting L-arginine and argininosuccinate synthetase 1 (ASS1).
ASS1 expression in HCC cell lines and primary hepatocytes was detected using polymerase chain reaction and western blotting. Proliferation, migration, signaling pathways, and nitric oxide production in HCC cell lines were measured using MTS, colony formation, wound healing, Western blot, and Griess assays.
ASS1 expression varied among the HCC cell lines, and cisplatin cytotoxicity was ASS1-dependent. L-arginine alone induced apoptosis in HCC cell lines, regardless of ASS1 expression; however, its effect was enhanced in ASS1-expressing HCC cell lines. Cisplatin cytotoxicity also increased, suggesting that L-arginine acts as a sensitizer to cisplatin in HCC cell lines. ASS1 and L-arginine produced nitric oxide and inhibited key proliferation- and survival-related signaling pathways such as PI3K/Akt and MAPK. Additionally, ASS1 and L-arginine reduced the expression of PKM1 and PKM2 in the glycolysis pathway.
Our study revealed that ASS1 and L-arginine exhibited anticancer effects in HCC and sensitized cisplatin-resistant HCC cells to chemotherapy. The combination of ASS1 and L-arginine significantly enhanced the anticancer effects, even in HCC cell lines with low or absent ASS1 expression. These findings highlight the critical roles of arginine and ASS1 in HCC and suggest that increasing arginine availability could be a promising therapeutic strategy.
背景/目的：肝细胞癌（HCC）是一种在全球发病率不断上升的恶性肿瘤。尽管已经做出了许多努力来确定HCC的有效治疗方法，但目前的策略仍存在局限性。我们提出了一种靶向L-精氨酸和精氨琥珀酸合成酶1（ASS1）的新方法。
使用聚合酶链反应和蛋白质印迹法检测HCC细胞系和原代肝细胞中ASS1的表达。使用MTS、集落形成、伤口愈合、蛋白质印迹和格里斯试验测量HCC细胞系中的增殖、迁移、信号通路和一氧化氮生成。
ASS1在HCC细胞系中的表达各不相同，顺铂的细胞毒性依赖于ASS1。单独的L-精氨酸可诱导HCC细胞系凋亡，无论ASS1表达如何；然而，其作用在表达ASS1的HCC细胞系中增强。顺铂的细胞毒性也增加，表明L-精氨酸在HCC细胞系中作为顺铂的增敏剂起作用。ASS1和L-精氨酸产生一氧化氮并抑制关键的增殖和生存相关信号通路，如PI3K/Akt和MAPK。此外，ASS1和L-精氨酸降低了糖酵解途径中PKM1和PKM2的表达。
我们的研究表明，ASS1和L-精氨酸在HCC中表现出抗癌作用，并使顺铂耐药的HCC细胞对化疗敏感。即使在ASS1表达低或无表达的HCC细胞系中，ASS1和L-精氨酸的联合使用也显著增强了抗癌作用。这些发现突出了精氨酸和ASS1在HCC中的关键作用，并表明增加精氨酸的可用性可能是一种有前景的治疗策略。
