Li Fenghua, Wang Yansa, Yan Ying
Department of Ophthalmology, Linyi People's Hospital, Linyi, Shandong 276000, P.R. China.
Exp Ther Med. 2017 May;13(5):2456-2462. doi: 10.3892/etm.2017.4252. Epub 2017 Mar 22.
The aim of the present study was to explore the effects of gambogenic acid (GNA) on the malignant behaviors of choroidal melanoma cells, including cell viability, cell cycle, migration and invasion, and to elucidate the underlying regulatory mechanism. The human choroidal melanoma cell line OCM-1 was treated with different concentrations of GNA and cell viability, colony formation ability, cell cycle, migration and invasion were analyzed. Additionally, cells were incubated with or without LY294002, a specific inhibitor of the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway, for 24 h. Levels of cell cycle-associated proteins (cyclin D1, cyclin E, cyclin-dependent kinase 2 and P21), epithelial-mesenchymal transition (EMT)-associated molecules (epithelial-cadherin, α-smooth muscle actin and vimentin) and phosphorylated (p)-AKT/AKT were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis. The results demonstrated that GNA significantly inhibited cell viability and induced cell cycle arrest at the G0/G1 phase in a dose-dependent manner (P<0.01). Furthermore, GNA administration significantly suppressed cell migration and invasion in a dose-dependent manner (P<0.01). Treatment with GNA or LY294002 induced a marked decrease in the expression of p-AKT/AKT, a significant downregulation in cell cycle-associated molecules (P<0.01), and a significant decrease in cell viability (P<0.01). Co-treatment with LY294002 and GNA had an additive effect on the growth of OCM-1 cells. In conclusion, the results of the present study suggest that treatment with GNA may inhibit cell viability and induce G0/G1 arrest. Furthermore, GNA may also inhibit cell metastasis via regulating EMT-associated molecules. The PI3K/Akt signaling pathway may be a key mechanism involved in the progression of choroidal melanoma, and GNA may serve as a potential therapeutic reagent for the treatment of this disease.
本研究的目的是探讨藤黄酸（GNA）对脉络膜黑色素瘤细胞恶性行为的影响，包括细胞活力、细胞周期、迁移和侵袭，并阐明其潜在的调控机制。用不同浓度的GNA处理人脉络膜黑色素瘤细胞系OCM-1，分析细胞活力、集落形成能力、细胞周期、迁移和侵袭。此外，将细胞与磷酸肌醇3激酶（PI3K）/蛋白激酶B（Akt）信号通路的特异性抑制剂LY294002一起孵育或不孵育24小时。使用逆转录定量聚合酶链反应和蛋白质印迹分析测定细胞周期相关蛋白（细胞周期蛋白D1、细胞周期蛋白E、细胞周期蛋白依赖性激酶2和P21）、上皮-间质转化（EMT）相关分子（上皮钙黏蛋白、α-平滑肌肌动蛋白和波形蛋白）以及磷酸化（p）-AKT/AKT的水平。结果表明，GNA以剂量依赖性方式显著抑制细胞活力并诱导细胞周期停滞在G0/G1期（P<0.01）。此外，给予GNA以剂量依赖性方式显著抑制细胞迁移和侵袭（P<0.01）。用GNA或LY294002处理导致p-AKT/AKT表达显著降低、细胞周期相关分子显著下调（P<0.01）以及细胞活力显著降低（P<0.01）。LY294002和GNA联合处理对OCM-1细胞的生长具有相加作用。总之，本研究结果表明，GNA处理可能抑制细胞活力并诱导G0/G1期停滞。此外，GNA还可能通过调节EMT相关分子来抑制细胞转移。PI3K/Akt信号通路可能是脉络膜黑色素瘤进展的关键机制，GNA可能作为治疗该疾病的潜在治疗试剂。
