Metastasis Suppressor NME1 Directly Activates Transcription of the ALDOC Gene in Melanoma Cells.

Metastasis Suppressor NME1 Directly Activates Transcription of the ALDOC Gene in Melanoma Cells.
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DOI:
10.21873/anticanres.12956
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发表时间:
2018-11
影响因子:
2
通讯作者:
Kaetzel DM
Kaetzel DM
中科院分区:
医学4区
文献类型:
--
作者:
Pamidimukkala NV;Leonard MK;Snyder D;McCorkle JR;Kaetzel DM

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NME/NM23 核苷二磷酸激酶 1 (NME1) 是一种转移抑制基因,在转移性癌症中表现出表达降低,并具有抑制癌细胞转移活性的能力。我们之前鉴定出 NME1 调控基因在人类黑色素瘤中具有预后价值。这项研究是在黑色素瘤细胞系中进行的,旨在阐明 NME 调节这些基因之一醛缩酶 C (ALDOC) 的机制。使用 qRT-PCR 和免疫印迹分析测量 ALDOC mRNA 和蛋白质表达。采用启动子荧光素酶构建体和染色质免疫沉淀来测量 NME1 对 ALDOC 转录的影响。 NME1 增强了 ALDOC 转录,这通过 ALDOC 前 mRNA 的表达增加和 ALDOC 启动子-荧光素酶模块的活性来证明。在 ALDOC 启动子处检测到 NME1,强制 NME1 表达导致 NME1 对启动子的占据增强,表观遗传激活标记(H3K4me3 和 H3K27ac)的存在增加,以及 RNA 聚合酶 II 的募集。这是第一项表明 NME1 通过直接结合靶基因启动子区域来诱导转录的研究。
NME/NM23 nucleoside dipho-sphate kinase 1 (NME1) is a metastasis suppressor gene, exhibiting reduced expression in metastatic cancers and the ability to suppress metastatic activity of cancer cells. We previously identified NME1-regulated genes with prognostic value in human melanoma. This study was conducted in melanoma cell lines aiming to elucidate the mechanism through which NME regulates one of these genes, aldolase C (ALDOC). ALDOC mRNA and protein expression was measured using qRT-PCR and immunoblot analyses. Promoter-luciferase constructs and chromatin immunoprecipitation were employed to measure the impact of NME1 on ALDOC transcription. NME1 enhanced ALDOC transcription, evidenced by increased expression of ALDOC pre-mRNA and activity of an ALDOC promoter-luciferase module. NME1 was detected at the ALDOC promoter, and forced NME1 expression resulted in enhanced occupancy of the promoter by NME1, increased presence of epigenetic activation markers (H3K4me3 and H3K27ac), and recruitment of RNA polymerase II. This is the first study to indicate that NME1 induces transcription through its direct binding to the promoter region of a target gene.
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