Department of Ophthalmology, College of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee, USA.
Invest Ophthalmol Vis Sci. 2010 Jul;51(7):3813-9. doi: 10.1167/iovs.09-5042.
PURPOSE. To investigate the effectiveness of a novel isoquinoline derivative, EDL-155, in killing retinoblastoma in vitro and in vivo. METHODS. Dose-response curves were generated in which Y79 retinoblastoma cells tagged with luciferase (Y79-Luc) were treated with serial concentrations of EDL-155. Electron microscopy was used to evaluate the ultrastructural morphology of EDL-155-treated Y79 cells. To determine whether autophagy was induced in EDL-155-treated Y79-Luc cells, staining with acridine orange and LC-3 immunoblot analysis was performed. To evaluate the efficacy of EDL-155 in vivo, Y79-Luc retinoblastoma cells were injected into the vitreous cavity of newborn rats, followed by periocular injections of EDL-155 (20 mg/kg/day) or an equivalent dosage of saline. RESULTS. EDL-155 appeared to destroy the retinoblastoma cells in vitro with an EC(50) of 9.1 micriM. EDL-155-treated retinoblastoma cells displayed a lack of viable mitochondria and the presence of autophagosomes wrapped in the characteristic double membranes. Acridine orange staining of EDL-155-treated retinoblastoma cells demonstrated the accumulation of vacuoles, and the immunoblots displayed a shift in molecular weight of LC-3, indicative of incorporation into autophagosome vesicles. In the retinoblastoma animal model, four doses of EDL-155 were delivered over 4 days, which was sufficient to see a significant decrease (P = 0.01) in viable intraocular tumors. Seven of the 25 rats treated with EDL-155 had no detectable living tumor. No significant decrease in viable tumor was observed in control animals. CONCLUSIONS. EDL-155 appears to eliminate retinoblastoma cells by disrupting mitochondria and inducing autophagy. Local delivery of EDL-155 may be an effective therapy for some types of ocular cancers.
目的。研究新型异喹啉衍生物 EDL-155 在体外和体内杀伤视网膜母细胞瘤的效果。
方法。生成剂量反应曲线，其中用 EDL-155 处理标记有荧光素酶的 Y79 视网膜母细胞瘤细胞（Y79-Luc）。用电子显微镜评估 EDL-155 处理的 Y79 细胞的超微结构形态。为了确定自噬是否在 EDL-155 处理的 Y79-Luc 细胞中被诱导，用吖啶橙染色和 LC-3 免疫印迹分析进行检测。为了评估 EDL-155 在体内的疗效，将 Y79-Luc 视网膜母细胞瘤细胞注射到新生大鼠的玻璃体腔中，然后在眼周注射 EDL-155（20mg/kg/天）或等量生理盐水。
结果。EDL-155 在体外似乎可以破坏视网膜母细胞瘤细胞，其 EC（50）为 9.1 微摩尔。EDL-155 处理的视网膜母细胞瘤细胞显示缺乏有活力的线粒体和存在自噬体，这些自噬体被特征性的双层膜包裹。吖啶橙染色的 EDL-155 处理的视网膜母细胞瘤细胞显示出空泡的积累，免疫印迹显示 LC-3 的分子量发生了转移，表明其已整合到自噬体囊泡中。在视网膜母细胞瘤动物模型中，在 4 天内给予 EDL-155 四剂，足以观察到活眼内肿瘤的显著减少（P=0.01）。25 只接受 EDL-155 治疗的大鼠中有 7 只没有检测到存活的肿瘤。对照动物中未观察到活肿瘤的显著减少。
结论。EDL-155 似乎通过破坏线粒体和诱导自噬来消除视网膜母细胞瘤细胞。EDL-155 的局部给药可能是治疗某些类型眼部癌症的有效方法。
