China Academy of Chinese Medical Sciences, Institute of Geriatric Medicine, Xiyuan Hospital, Beijing, People's Republic of China.
Department of Hematology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, People's Republic of China.
Drug Des Devel Ther. 2020 Apr 30;14:1641-1650. doi: 10.2147/DDDT.S239158. eCollection 2020.
Previous studies have shown that DNA methylation plays a significant role in myelodysplastic syndrome (MDS). In addition to hypermethylation, aberrant hypomethylation can result in the transcriptional activation of oncogenes in cancer, including MDS. Therefore, drugs targeting DNA hypomethylation are needed for the treatment of MDS. This study aimed to investigate whether AsS promoted hypomethylation by increasing DNA methyltransferases (DNMTs) expression in MDS.
Ten bone marrow samples from MDS patients and 3 healthy donors were obtained for the examination of the DNA methylation with a Human Methylation 850K BeadChip. The mRNA expressions for the DNMTs in the ten MDS patients and 3 controls were compared by Q-PCR. Then, the MDS cell line SKM-1 was treated with AsS. After 2 days of treatment, Human Methylation 850K BeadChip was applied to analyze the changes of gene methylation status in the cells. Q-PCR and Western blot were taken to test the changes of mRNA and protein expressions for DNMTs in SKM-1 cells after treatment.
Five hundred ninety-two abnormally hypomethylated genes were found in MDS patients compared to those in controls by Human Methylation 850K. The mRNA expressions of DNMTs (DNMT1, DNMT3a and DNMT3b) in MDS patients were significantly lower than those in healthy individuals. The IC50 value of AsS for SKM-1 cells was 4.97 μmol/L.Treatment with AsS at 2 μmoL/L resulted in significant alterations in the methylation levels at 1718 sites in SKM-1 cells compared to those in the controls. Hypermethylation was observed in 1625 sites (94.58%), corresponding to 975 genes, compared to those in the controls. Finally, the expression levels of DNMTs (DNMT1, DNMT3a, and DNMT3b) significantly increased in SKM-1 cells treated with AsS at 2 μmoL/L and 4 μmoL/L.
These data show a potential clinical application of AsS as an innovative hypermethylation agent in MDS.
先前的研究表明，DNA 甲基化在骨髓增生异常综合征（MDS）中起着重要作用。除了超甲基化，异常的低甲基化可导致癌症中癌基因的转录激活，包括 MDS。因此，需要针对 DNA 低甲基化的药物来治疗 MDS。本研究旨在探讨 AsS 是否通过增加 DNA 甲基转移酶（DNMTs）的表达来促进 MDS 中的低甲基化。
从 10 例 MDS 患者和 3 例健康供体的骨髓样本中获得 DNA 甲基化的检测，采用 Human Methylation 850K BeadChip。通过 Q-PCR 比较 10 例 MDS 患者和 3 例对照的 DNMTs mRNA 表达。然后用 AsS 处理 MDS 细胞系 SKM-1。治疗 2 天后，应用 Human Methylation 850K BeadChip 分析细胞中基因甲基化状态的变化。通过 Q-PCR 和 Western blot 检测治疗后 SKM-1 细胞中 DNMTs 的 mRNA 和蛋白表达的变化。
通过 Human Methylation 850K 发现 MDS 患者与对照组相比有 592 个异常低甲基化基因。MDS 患者的 DNMTs（DNMT1、DNMT3a 和 DNMT3b）mRNA 表达明显低于健康个体。AsS 对 SKM-1 细胞的 IC50 值为 4.97 μmol/L。用 2 μmoL/L 的 AsS 处理 SKM-1 细胞，与对照组相比，SKM-1 细胞中的 1718 个位点的甲基化水平发生显著改变。与对照组相比，观察到 1625 个位点（94.58%）发生过度甲基化，对应 975 个基因。最后，用 2 μmoL/L 和 4 μmoL/L 的 AsS 处理 SKM-1 细胞后，DNMTs（DNMT1、DNMT3a 和 DNMT3b）的表达水平明显升高。
这些数据表明 AsS 作为 MDS 中一种创新的高甲基化剂具有潜在的临床应用价值。
