Liu Yiheng, Liu Qingqing, Huang Tianyi, Zhang Shengjie, Zhou Zicheng, Gu Chiyu, Lu Cuihua
Department of Gastroenterology, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, 226001, China.
Department of Emergency, Nantong Third People's Hospital, Affiliated Nantong Hospital 3 of Nantong University, Nantong, 226001, Jiangsu, China.
Dig Dis Sci. 2025 Apr 16. doi: 10.1007/s10620-025-08916-0.
Hepatocellular carcinoma (HCC) is one of the primary liver tumors with high incidence and mortality. RNA G-quadruplexes (rG4) are nucleic acid structures involved in gene expression and genome duplication. rG4 exerts its function by interacting with rG4-binding proteins. The carboxypyridostatin (cPDS), a specific ligand of rG4, are widely studied in numerous tumors. However, the role of cPDS in HCC and its regulatory mechanisms are not yet fully understood. Our study aimed to discuss the regulatory mode of cPDS on Baculoviral IAP Repeat Containing 3 (BIRC3) expression and its impact on proliferation, apoptosis, and other biologic functions in HCC.
We conducted colony formation, CCK8, Edu incorporation, scratch healing, and cell spheroid formation assays to analyze the function of cPDS on cell proliferation and migration. Additionally, we explored the role of cPDS in regulating BIRC3 expression by Western blot and qRT-PCR. Furthermore, we evaluated the impact of BIRC3 on cell proliferation and subcutaneous tumor formation in nude mice. Finally, we analyzed the regulatory mechanisms of cPDS on cell apoptosis by Western blot, qRT-PCR, flow cytometry, and Annexin V-FITC staining.
Our results demonstrated that cPDS inhibited HCC cells proliferation and migration. Moreover, cPDS elevated the mRNA level while inhibiting the protein expression of BIRC3 in HCC cells. Overexpression of BIRC3 significantly enhanced the proliferation of HCC cells. In the nude mice model, BIRC3 significantly increased the tumor volume and weight. Mechanistically, cPDS promoted cell apoptosis via inhibiting BIRC3-mediated anti-apoptotic effect.
Our findings revealed a critical role of rG4 ligand cPDS in HCC progression and indicate that cPDS may be used for HCC treatment considering its tumor inhibitory properties by regulating cell apoptosis.
肝细胞癌（HCC）是发病率和死亡率较高的原发性肝脏肿瘤之一。RNA G-四链体（rG4）是参与基因表达和基因组复制的核酸结构。rG4通过与rG4结合蛋白相互作用发挥其功能。羧基吡啶他汀（cPDS）作为rG4的特异性配体，在众多肿瘤中得到广泛研究。然而，cPDS在HCC中的作用及其调控机制尚未完全明确。我们的研究旨在探讨cPDS对含杆状病毒IAP重复序列3（BIRC3）表达的调控模式及其对HCC细胞增殖、凋亡和其他生物学功能的影响。
我们进行了集落形成、CCK8、Edu掺入、划痕愈合和细胞球形成实验，以分析cPDS对细胞增殖和迁移的作用。此外，我们通过蛋白质免疫印迹法（Western blot）和实时定量逆转录聚合酶链反应（qRT-PCR）探讨了cPDS在调节BIRC3表达中的作用。此外，我们评估了BIRC3对裸鼠细胞增殖和皮下肿瘤形成的影响。最后，我们通过蛋白质免疫印迹法、qRT-PCR、流式细胞术和膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）染色分析了cPDS对细胞凋亡的调控机制。
我们的结果表明，cPDS抑制HCC细胞的增殖和迁移。此外，cPDS提高了HCC细胞中BIRC3的mRNA水平，同时抑制其蛋白表达。BIRC3的过表达显著增强了HCC细胞的增殖。在裸鼠模型中，BIRC3显著增加了肿瘤体积和重量。机制上，cPDS通过抑制BIRC3介导的抗凋亡作用促进细胞凋亡。
我们的研究结果揭示了rG4配体cPDS在HCC进展中的关键作用，并表明考虑到cPDS通过调节细胞凋亡具有肿瘤抑制特性，其可能用于HCC治疗。
