组蛋白 H3 赖氨酸 4 的低甲基化和组蛋白 H3 赖氨酸 27 的三甲基化/高甲基化作为原发性中枢神经系统淋巴瘤患者预后不良的表观遗传学标志物。
Hypo-trimethylation of Histone H3 Lysine 4 and Hyper-tri/dimethylation of Histone H3 Lysine 27 as Epigenetic Markers of Poor Prognosis in Patients with Primary Central Nervous System Lymphoma.
机构信息
Division of Neuro-Oncology and Department of Neurosurgery, Samsung Changwon Hospital, Sungkyunkwan University School of Medicine, Changwon, Korea.
Department of New Biology, Daegu Gyeongbuk Institute of Science and Technology, Daegu, Korea.
出版信息
Cancer Res Treat. 2022 Jul;54(3):690-708. doi: 10.4143/crt.2021.1121. Epub 2021 Nov 17.
PURPOSE
This study aimed to investigate the methylation status of major histone modification sites in primary central nervous system lymphoma (PCNSL) samples and examine their prognostic roles in patients with PCNSL.
MATERIALS AND METHODS
Between 2007 and 2020, 87 patients were histopathologically diagnosed with PCNSL. We performed immunohistochemical staining of the formalin-fixed paraffin-embedded samples of PCNSL for major histone modification sites, such as H3K4, H3K9, H3K27, H3K14, and H3K36. After detection of meaningful methylation sites, we examined histone modification enzymes that induce methylation or demethylation at each site using immunohistochemical staining. The meaningful immunoreactivity was validated by western blotting using fresh tissue of PCNSL.
RESULTS
More frequent recurrences were found in hypomethylation of H3K4me3 (p=0.004) and hypermethylation of H3K27me2 (p<0.001) and H3K27me3 (p=0.002). These factors were also statistically related to short PFS and overall survival in the univariate and multivariate analyses. Next, histone modification enzymes inducing the demethylation of H3K4 (lysine-specific demethylase-1/2 and Jumonji AT-rich interactive domain [JARID] 1A-D]) and methylation of H3K27 (enhancer of zeste homolog [EZH]-1/2) were immu- nohistochemically stained. Among them, the immunoreactivity of JARID1A inversely associated with the methylation status of H3K4me3 (R2=-1.431), and immunoreactivity of EZH2 was directly associated with the methylation status of H3K27me2 (R2=0.667) and H3K27me3 (R2=0.604). These results were validated by western blotting in fresh PCNSL samples.
CONCLUSION
Our study suggests that hypomethylation of H3K4me3 and hypermethylation of H3K27me2 and H3K27me3 could be associated with poor outcomes in patients with PCNSL and that these relationships are modified by JARID1A and EZH2.
目的
本研究旨在探讨原发性中枢神经系统淋巴瘤（PCNSL）样本中主要组蛋白修饰位点的甲基化状态，并研究其在 PCNSL 患者中的预后作用。
材料和方法
在 2007 年至 2020 年期间，87 例患者经组织病理学诊断为 PCNSL。我们对 PCNSL 的福尔马林固定石蜡包埋样本进行了主要组蛋白修饰位点（如 H3K4、H3K9、H3K27、H3K14 和 H3K36）的免疫组织化学染色。在检测到有意义的甲基化位点后，我们使用免疫组织化学染色法检测了诱导每个位点甲基化或去甲基化的组蛋白修饰酶。使用新鲜的 PCNSL 组织对有意义的免疫反应性进行了 Western 印迹验证。
结果
H3K4me3 低甲基化（p=0.004）和 H3K27me2（p<0.001）和 H3K27me3（p=0.002）高甲基化的患者复发更为频繁。这些因素在单因素和多因素分析中也与较短的 PFS 和总生存期相关。接下来，我们对诱导 H3K4 去甲基化（赖氨酸特异性去甲基酶-1/2 和 JARID1A-D）和 H3K27 甲基化（增强子的锌指蛋白[EZH]-1/2）的组蛋白修饰酶进行了免疫组织化学染色。其中，JARID1A 的免疫反应性与 H3K4me3 的甲基化状态呈负相关（R2=-1.431），EZH2 的免疫反应性与 H3K27me2（R2=0.667）和 H3K27me3（R2=0.604）的甲基化状态呈正相关。这些结果在新鲜的 PCNSL 样本的 Western 印迹中得到了验证。
结论
我们的研究表明，H3K4me3 的低甲基化和 H3K27me2 和 H3K27me3 的高甲基化可能与 PCNSL 患者的不良预后相关，并且这些关系受 JARID1A 和 EZH2 的调节。
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