Department of Ophthalmology, The Second Clinical Medical College (Shenzhen People's Hospital), Jinan University, Shenzhen, 518020, People's Republic of China.
Department of Obstetrics and Gynecology, The Second Clinical Medical College (Shenzhen People's Hospital), Jinan University, Shenzhen, 518020, People's Republic of China.
J Transl Med. 2017 Dec 29;15(1):268. doi: 10.1186/s12967-017-1372-8.
In our previous study, we revealed that MEG3 was a tumor suppressor gene in retinoblastoma and inhibited proliferation of retinoblastoma cells by regulating the activity of the Wnt/β-catenin pathway. Here, we further explored the mechanism of MEG3 inactivation in retinoblastoma.
MSP and qRT-PCR were performed to detect the methylation status of MEG3 promoter and levels of MEG3 expression, respective. To further explore relationship between MEG3 expression and epigenetic modifications, 5-Aza-CdR was used to interfere with DNA methylation. In addition, we evaluated proliferation, apoptosis and the expression of β-catenin via CCK-8, flow cytometric analysis and western blot analysis, respective.
Hypermethylation of MEG3 promoter was observed more frequently in retinoblastoma tissues and was highly associated with low MEG3 expression and poor survival of retinoblastoma patients. We also provided evidence demonstrating that hypermethylation of MEG3 promoter depressed MEG3 expression, promoted proliferation, inhibited apoptosis and increased β-catenin expression of retinoblastoma cells in vitro.
Our present study indicates that promoter silencing by hypermethylation may account for the loss of MEG3 expression and predict poor prognosis.
在我们之前的研究中，我们揭示了 MEG3 是视网膜母细胞瘤中的肿瘤抑制基因，通过调节 Wnt/β-catenin 通路的活性来抑制视网膜母细胞瘤细胞的增殖。在这里，我们进一步探讨了 MEG3 在视网膜母细胞瘤中失活的机制。
采用 MSP 和 qRT-PCR 分别检测 MEG3 启动子的甲基化状态和 MEG3 表达水平。为了进一步探讨 MEG3 表达与表观遗传修饰之间的关系，使用 5-Aza-CdR 干扰 DNA 甲基化。此外，我们通过 CCK-8、流式细胞术分析和 Western blot 分析分别评估增殖、凋亡和β-catenin 的表达。
在视网膜母细胞瘤组织中观察到 MEG3 启动子的高甲基化频率更高，与低 MEG3 表达和视网膜母细胞瘤患者的不良预后高度相关。我们还提供了证据表明，MEG3 启动子的高甲基化抑制了 MEG3 的表达，促进了视网膜母细胞瘤细胞的增殖，抑制了凋亡，并增加了 β-catenin 的表达。
本研究表明，启动子的沉默可能是由于 MEG3 表达的缺失，并预测预后不良。
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