He Haifa, Li Yin, Wang Yuan, Li Man
Department of Pathology, Nanyang Central Hospital, No.312 Gongnong Road, Wancheng District, Nanyang, 473005, Henan, China.
Department of Teaching and Research Section of Pathology, Nanyang Medical College, Nanyang, 473000, China.
Biochem Genet. 2025 Feb 18. doi: 10.1007/s10528-025-11030-5.
Non-small cell lung cancer (NSCLC) is the leading cause of cancer death worldwide. Geniposide, an active compound of Gardeniae Fructus, has antithrombotic, antitumor, neuroprotective, hepatoprotective, cholestatic, and other effects. The present study aimed to investigate the effects of geniposide on NSCLC cells, as well as its underlying mechanism. Two NSCLC cell lines (H1975 and A549) were treated with different doses of geniposide. The proliferation, apoptosis, migratory and invasive capacities, epithelial-mesenchymal transition (EMT), and stem cell characteristics of NSCLC cells were evaluated using a series of in vitro experiments, including colony formation, flow cytometry, wound healing, transwell, western blotting, and tube formations assays. H1975 cells were subcutaneously injected into nude mice to establish the xenograft tumor models, and the models were intraperitoneally injected with 100 mg/kg geniposide or/and 6 mg/kg SKL2001, an agonist of Wnt pathway. Immunohistochemistry, immunofluorescence, and western blotting analyses of the tumors were performed. Geniposide restrained the proliferation of NSCLC cells, as shown by reduced number of colonies and downregulation of Ki67 and PCNA expression levels. Geniposide promoted apoptosis by reducing Bcl-2 expression and increasing Bax expression. Additionally, geniposide inhibited the migratory and invasive abilities of NSCLC cells as well as reversed the EMT by downregulating vimentin, N-cadherin, snail, and slug and upregulating E-cadherin in the absence or presence of TGF-β1. Furthermore, geniposide attenuated the stem cell characteristics of NSCLC cells. In mechanism, geniposide repressed the activation of Wnt/β-catenin pathway. SKL2001 reversed the anti-NSCLC effects of geniposide in vitro. In the xenograft tumor models, 100 mg/kg geniposide suppressed NSCLC tumor growth, which was reversed by SKL2001 treatment. Overall, geniposide inhibits NSCLC progression by reducing cancer cell proliferation, migration, invasiveness, EMT, and stem cell characteristics. This information might provide novel insights into the potential use of geniposide in lung cancer intervention.
非小细胞肺癌（NSCLC）是全球癌症死亡的主要原因。栀子苷是栀子的一种活性化合物，具有抗血栓形成、抗肿瘤、神经保护、肝脏保护、胆汁淤积等作用。本研究旨在探讨栀子苷对NSCLC细胞的影响及其潜在机制。用不同剂量的栀子苷处理两种NSCLC细胞系（H1975和A549）。使用一系列体外实验，包括集落形成、流式细胞术、伤口愈合、Transwell、蛋白质印迹和管形成实验，评估NSCLC细胞的增殖、凋亡、迁移和侵袭能力、上皮-间质转化（EMT）和干细胞特性。将H1975细胞皮下注射到裸鼠体内建立异种移植肿瘤模型，并给模型腹腔注射100 mg/kg栀子苷或/和6 mg/kg SKL2001（Wnt途径激动剂）。对肿瘤进行免疫组织化学、免疫荧光和蛋白质印迹分析。栀子苷抑制NSCLC细胞的增殖，表现为集落数量减少以及Ki67和PCNA表达水平下调。栀子苷通过降低Bcl-2表达和增加Bax表达促进凋亡。此外，在存在或不存在转化生长因子-β1（TGF-β1）的情况下，栀子苷通过下调波形蛋白、N-钙黏蛋白、蜗牛蛋白和蛞蝓蛋白并上调E-钙黏蛋白，抑制NSCLC细胞的迁移和侵袭能力，并逆转EMT。此外，栀子苷减弱了NSCLC细胞的干细胞特性。机制上，栀子苷抑制Wnt/β-连环蛋白途径的激活。SKL2001在体外逆转了栀子苷的抗NSCLC作用。在异种移植肿瘤模型中，100 mg/kg栀子苷抑制NSCLC肿瘤生长，而SKL2001处理可逆转这一作用。总体而言，栀子苷通过降低癌细胞增殖、迁移、侵袭、EMT和干细胞特性来抑制NSCLC进展。这些信息可能为栀子苷在肺癌干预中的潜在应用提供新的见解。
