Dr Rath Research Institute, Cancer Division, Santa Clara, CA 95050, USA.
Oncol Rep. 2013 Jan;29(1):29-33. doi: 10.3892/or.2012.2110. Epub 2012 Oct 29.
Retinoblastoma is one of the most common ocular malignancies in children under the age of six. Occasionally, retinoblastoma metastasizes to extraocular organs including the bone, lung and brain. Left untreated, retinoblastoma is fatal. At present, there is no effective treatment for metastatic retinoblastoma. We investigated the antineoplastic activity of a nutrient mixture (NM) (lysine, proline, ascorbic acid and green tea extract) at concentrations of 10, 50, 100, 500 and 1,000 µg/ml in triplicate at each dose in the human malignant retinoblastoma Y-79 cell line. The parameters used were cell proliferation, expression of matrix metalloproteinases (MMPs), invasion through Matrigel, morphology and apoptosis. Cell viability was assessed by trypan blue dye exclusion test. Invasion was evaluated through Matrigel and MMP activity by gelatinase zymography. H&E staining for morphological cell alterations and apoptotic studies using the Live Green Poly Caspase Detection kit were also conducted. The nutrient mixture at 10-100 µg/ml demonstrated approximately 25% toxicity towards Y-79 retinoblastoma cells and significant toxicity at 500 and 1,000 µg/ml. The Y-79 cells secreted only MMP-2 as demonstrated by zymography; the nutrient mixture had no effect on MMP-2 expression up to 100 µg/ml, but completely blocked it at 500 µg/ml. Importantly, Y-79 retinoblastoma cells were not invasive through Matrigel. H&E staining showed cell morphological changes related to apoptosis, which was confirmed using the Live Green Poly Caspase Detection kit. Our results suggest that this nutrient mixture, which inhibited cell proliferation, expression of MMP-2 and induced apoptosis, may be a candidate for further exploration for its therapeutic potential in metastatic retinoblastoma.
视网膜母细胞瘤是 6 岁以下儿童中最常见的眼部恶性肿瘤之一。偶尔，视网膜母细胞瘤会转移到眼外器官，包括骨骼、肺和脑。如果不治疗，视网膜母细胞瘤是致命的。目前，转移性视网膜母细胞瘤没有有效的治疗方法。我们研究了一种营养混合物（NM）（赖氨酸、脯氨酸、抗坏血酸和绿茶提取物）在浓度为 10、50、100、500 和 1000μg/ml 时对人恶性视网膜母细胞瘤 Y-79 细胞系的抗肿瘤活性，每个剂量重复三次。使用的参数是细胞增殖、基质金属蛋白酶（MMPs）表达、通过 Matrigel 的侵袭、形态和细胞凋亡。细胞活力通过台盼蓝排斥试验评估。通过 Matrigel 和明胶酶谱法评估 MMP 活性来评估侵袭。还进行了 H&E 染色以评估形态学细胞改变和使用 Live Green Poly Caspase Detection 试剂盒进行的凋亡研究。营养混合物在 10-100μg/ml 时对 Y-79 视网膜母细胞瘤细胞表现出约 25%的毒性，在 500μg/ml 和 1000μg/ml 时表现出显著毒性。Y-79 细胞仅分泌 MMP-2，如明胶酶谱所示；营养混合物在高达 100μg/ml 时对 MMP-2 表达没有影响，但在 500μg/ml 时完全阻断了它。重要的是，Y-79 视网膜母细胞瘤细胞不能通过 Matrigel 侵袭。H&E 染色显示与凋亡相关的细胞形态变化，这一点通过使用 Live Green Poly Caspase Detection 试剂盒得到了证实。我们的结果表明，这种营养混合物抑制细胞增殖、MMP-2 表达并诱导细胞凋亡，可能是进一步探索其在转移性视网膜母细胞瘤治疗潜力的候选药物。
