Coordinated transcription of ANRIL and P16 genes is silenced by P16 DNA methylation
Coordinated transcription of ANRIL and P16 genes is silenced by P16 DNA methylation
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ANRIL 和 P16 基因的协调转录因 P16 DNA 甲基化而沉默。
DOI：
10.21147/j.issn.1000-9604.2018.01.10
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发表时间：
2018-02-01
影响因子：
5.1
通讯作者：
Deng, Dajun
中科院分区：
文献类型：
--
作者：
Gan, Ying;Ma, Wanru;Deng, Dajun
目的：为探讨ANRIL与P15、P14、P16基因在同一位点转录的关系及其调控机制，采用生物信息学方法，利用CCLE数据库进行分析。变性高效液相色谱法（DHPLC）检测CpG岛甲基化。使用定量实时聚合酶链反应（qRTPCR）测定基因转录水平。结果：ANRIL的表达水平与P16的表达水平呈正相关，与P15的表达水平无相关性。这在人细胞系和患者结肠组织样品中得到证实。此外，ANRIL在结肠癌组织中显著上调。ANRIL和P16的转录仅在P16等位基因未甲基化的细胞系中观察到，而在P16等位基因完全甲基化的细胞系中未观察到。值得注意的是，P16特异性甲基化显著降低了BGC 823和GES 1细胞中P16和ANRIL的转录。相反，P16特异性去甲基化重新激活H1299细胞中ANRIL和P16的转录（P
Objective: To investigate the relationship between the transcription of ANRIL, P15, P14 and P16 at the same locus and the regulation mechanism of ANRIL.Methods: Publicly available database of Cancer Cell Line Encyclopedia (CCLE) was used in bioinformatic analyses. Methylation of CpG islands was detected by denaturing high performance liquid chromatography (DHPLC). Gene transcript levels were determined using quantitative real-time polymerase chain reaction (qRTPCR) assays. An engineered P16-specific transcription factor and DNA methyltransferase were used to induce P16specific DNA demethylation and methylation.Results: The expression level of ANRIL was positively and significantly correlated with that of P16 but not with that of P15 in the CCLE database. This was confirmed in human cell lines and patient colon tissue samples. In addition, ANRIL was significantly upregulated in colon cancer tissues. Transcription of ANRIL and P16 was observed only in cell lines in which the P16 alleles were unmethylated and not in cell lines with fully methylated P16 alleles. Notably, P16-specific methylation significantly decreased transcription of P16 and ANRIL in BGC823 and GES1 cells. In contrast, P16-specific demethylation re-activated transcription of ANRIL and P16 in H1299 cells (P