Tao Yan-Fang, Xu Li-Xiao, Lu Jun, Cao Lan, Li Zhi-Heng, Hu Shao-Yan, Wang Na-Na, Du Xiao-Juan, Sun Li-Chao, Zhao Wen-Li, Xiao Pei-Fang, Fang Fang, Li Yan-Hong, Li Gang, Zhao He, Li Yi-Ping, Xu Yun-Yun, Ni Jian, Wang Jian, Feng Xing, Pan Jian
Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
J Transl Med. 2014 Jun 25;12:182. doi: 10.1186/1479-5876-12-182.
Acute myeloid leukemia (AML) is the second most common form of leukemia in children. Aberrant DNA methylation patterns are a characteristic feature in various tumors, including AML. Metallothionein III (MT3) is a tumor suppresser reported to show promoter hypermethylated in various cancers. However, the expression and molecular function of MT3 in pediatric AML is unclear.
Eleven human leukemia cell lines and 41 pediatric AML samples and 20 NBM/ITP (Norma bone marrow/Idiopathic thrombocytopenic purpura) control samples were analyzed. Transcription levels of MT3 were evaluated by semi-quantitative and real-time PCR. MT3 methylation status was determined by methylation specific PCR (MSP) and bisulfite genomic sequencing (BSG). The molecular mechanism of MT3 was investigated by apoptosis assays and PCR array analysis.
The MT3 promoter was hypermethylated in leukemia cell lines. More CpG's methylated of MT3 was observed 39.0% pediatric AML samples compared to 10.0% NBM controls. Transcription of MT3 was also significantly decreased in AML samples compared to NBM/ITP controls (P < 0.001); patients with methylated MT3 exhibited lower levels of MT3 expression compared to those with unmethylated MT3 (P = 0.049). After transfection with MT3 lentivirus, proliferation was significantly inhibited in AML cells in a dose-dependent manner (P < 0.05). Annexin V assay showed that apoptosis was significantly upregulated MT3-overexpressing AML cells compared to controls. Real-time PCR array analysis revealed 34 dysregulated genes that may be implicated in MT3 overexpression and apoptosis in AML, including FOXO1.
MT3 may be a putative tumor suppressor gene in pediatric AML. Epigenetic inactivation of MT3 via promoter hypermethylation was observed in both AML cell lines and pediatric AML samples. Overexpression of MT3 may inhibit proliferation and induce apoptosis in AML cells. FOXO1 was dysregulated in MT3-overexpressing cells, offering an insight into the mechanism of MT3-induced apoptosis. However, further research is required to determine the underlying molecular details.
急性髓系白血病（AML）是儿童中第二常见的白血病形式。异常的DNA甲基化模式是包括AML在内的各种肿瘤的一个特征性表现。金属硫蛋白III（MT3）是一种肿瘤抑制因子，据报道在各种癌症中其启动子存在高甲基化。然而，MT3在儿童AML中的表达及分子功能尚不清楚。
对11种人类白血病细胞系、41份儿童AML样本和20份NBM/ITP（正常骨髓/特发性血小板减少性紫癜）对照样本进行分析。通过半定量和实时PCR评估MT3的转录水平。通过甲基化特异性PCR（MSP）和亚硫酸氢盐基因组测序（BSG）确定MT3的甲基化状态。通过凋亡分析和PCR阵列分析研究MT3的分子机制。
白血病细胞系中MT3启动子存在高甲基化。与10.0%的NBM对照相比，在39.0%的儿童AML样本中观察到更多MT3的CpG位点发生甲基化。与NBM/ITP对照相比，AML样本中MT3的转录也显著降低（P < 0.001）；与未甲基化MT3的患者相比，甲基化MT3的患者MT3表达水平较低（P = 0.049）。用MT3慢病毒转染后，AML细胞的增殖以剂量依赖方式受到显著抑制（P < 0.05）。膜联蛋白V分析显示，与对照相比，MT3过表达的AML细胞凋亡显著上调。实时PCR阵列分析揭示了34个失调基因，这些基因可能与AML中MT3过表达和凋亡有关，包括FOXO1。
MT3可能是儿童AML中的一个假定肿瘤抑制基因。在AML细胞系和儿童AML样本中均观察到通过启动子高甲基化导致MT3的表观遗传失活。MT3的过表达可能抑制AML细胞的增殖并诱导其凋亡。FOXO1在MT3过表达细胞中失调，为MT3诱导凋亡的机制提供了见解。然而，需要进一步研究以确定潜在的分子细节。
Zotero 插件
