Department of Ophthalmology, The Third Xiangya Hospital, Central South University, No. 138 Tongzipo Road, Yuelu District, Changsha City, Hunan Province, 430013, China.
Naunyn Schmiedebergs Arch Pharmacol. 2024 Feb;397(2):1003-1013. doi: 10.1007/s00210-023-02646-8. Epub 2023 Aug 9.
Retinoblastoma is a prevalent pediatric intraocular tumor. The suppressive effect of gentiopicroside (GPS) has been reported on various tumors. This study sought to determine the effect of GPS on retinoblastoma cell proliferation, apoptosis, invasion, and epithelial-mesenchymal transition (EMT), and tumorigenesis in nude mice. The effect and mechanism of GPS on growth, apoptosis, invasion, and EMT were determined by cell counting kit-8 (CCK-8), western blot, flow cytometry, and transwell assays in retinoblastoma cells. Y79 cells were injected into the vitreous cavity of BALB/c‑nude mice to construct a retinoblastoma mouse model. Tumor growth and mouse weight were monitored for sequential 5 weeks. The effect of GPS in vivo was assessed by immunohistochemistry (IHC), terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL), and western blot assays. GPS decreased the cell viability of both Y79 and Weri-Rb1 cells with the IC50 of 18.85 μM and 27.57 μM, respectively. Besides, GPS reduced the relative expression of proteins involved in proliferation and EMT, and the number of invading cells, while increased the apoptosis rate and the relative expressions of apoptosis proteins in retinoblastoma cells. Mechanically, GPS decreased the relative protein level of PI3K/AKT pathway, which was then recovered after 740 Y-P was applied. Correspondingly, 740 Y-P reversed the inhibitory effect of GPS on growth, invasion, and EMT, and the increased effect of GPS on apoptosis. Additionally, GPS decreased tumor volume and weight as well as the relative level of Ki-67, VEGF, p-PI3K/PI3K, and p-AKT/AKT, while increased the apoptosis rate in vivo. GPS inhibited retinoblastoma cell proliferation and invasion via deactivating the PI3K/AKT pathway in both cell and animal models.
视网膜母细胞瘤是一种常见的小儿眼内肿瘤。环烯醚萜苷(GPS)已被报道对多种肿瘤具有抑制作用。本研究旨在探讨 GPS 对视网膜母细胞瘤细胞增殖、凋亡、侵袭和上皮-间充质转化(EMT)以及裸鼠肿瘤发生的影响。通过细胞计数试剂盒-8(CCK-8)、蛋白质印迹、流式细胞术和 Transwell 测定法,在视网膜母细胞瘤细胞中确定 GPS 对生长、凋亡、侵袭和 EMT 的影响和机制。将 Y79 细胞注入 BALB/c-裸鼠玻璃体腔构建视网膜母细胞瘤小鼠模型。连续 5 周监测肿瘤生长和小鼠体重。通过免疫组织化学(IHC)、末端脱氧核苷酸转移酶 dUTP 缺口末端标记(TUNEL)和蛋白质印迹法评估 GPS 在体内的作用。GPS 降低了 Y79 和 Weri-Rb1 细胞的细胞活力,其 IC50 分别为 18.85 μM 和 27.57 μM。此外,GPS 降低了增殖和 EMT 相关蛋白的相对表达量和侵袭细胞数量,同时增加了视网膜母细胞瘤细胞的凋亡率和凋亡蛋白的相对表达量。在机制上,GPS 降低了 PI3K/AKT 通路的相对蛋白水平,而应用 740 Y-P 后则恢复。相应地,740 Y-P 逆转了 GPS 对生长、侵袭和 EMT 的抑制作用,以及 GPS 对凋亡的增强作用。此外,GPS 降低了肿瘤体积和重量以及 Ki-67、VEGF、p-PI3K/PI3K 和 p-AKT/AKT 的相对水平,同时增加了体内的凋亡率。GPS 通过在细胞和动物模型中失活 PI3K/AKT 通路抑制视网膜母细胞瘤细胞增殖和侵袭。