ALKBH5 promotes invasion and metastasis of gastric cancer by decreasing methylation of the lncRNA NEAT1
ALKBH5 promotes invasion and metastasis of gastric cancer by decreasing methylation of the lncRNA NEAT1
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DOI：
10.1007/s13105-019-00690-8
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发表时间：
2019-08-01
影响因子：
3.4
通讯作者：
Zhao, Yan
中科院分区：
文献类型：
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作者：
Zhang, Jun;Guo, Shuai;Zhao, Yan
N-6-甲基腺苷(m(6)A)是最常见的RNA转录后修饰，在肿瘤发病机制中发挥重要作用。然而，长非编码RNA(LncRNA)甲基化的生物学功能仍不清楚。作为一种去甲基酶，ALKBH5(烷基化修复同源蛋白5)参与了甲基化逆转。本研究旨在探讨LncRNA m(6)A修饰及其在胃癌中的作用。生物信息学预测了ALKBH5与lncRNAs的相互作用。采用基因沉默、RT-PCR、核质组分分离、刮片运动试验和跨孔迁移试验等5种方法对核旁斑点组装转录本1(NEAT1)的功能进行了研究。采用m(6)A RNA免疫沉淀和免疫荧光法检测胃癌细胞中NEAT1的甲基化状态。采用补救试验确定NEAT1和ALKBH5之间的关系。NEAT1是ALKBH5的一个潜在结合的lncRNA。NEAT1在GC细胞和组织中高表达。进一步的实验证实，NEAT1基因的敲除显著抑制了GC细胞的侵袭和转移。ALKBH5影响NEAT1的m(6)A水平。ALKBH5和NEAT1的结合影响EZH2(多梳抑制复合体的一个亚单位)的表达，从而影响胃癌的侵袭和转移。我们的发现表明了ALKBH5通过去甲基化lncRNA NEAT1促进胃癌侵袭和转移的新机制。它们可能是GC潜在的治疗靶点。
N-6-Methyladenosine (m(6)A) is the most common posttranscriptional modification of RNA and plays critical roles in cancer pathogenesis. However, the biological function of long noncoding RNA (lncRNA) methylation remains unclear. As a demethylase, ALKBH5 (alkylation repair homolog protein 5) is involved in mediating methylation reversal. The purpose of this study was to investigate lncRNA m(6)A modification and its role in gastric cancer (GC). Bioinformatics predicted interactions of ALKBH5 with lncRNAs. Five methods were employed to assess the function of nuclear paraspeckle assembly transcript 1 (NEAT1), including gene silencing, RT-PCR, separation of nuclear and cytoplasmic fractions, scrape motility assays, and transwell migration assays. Then, m(6)A RNA immunoprecipitation and immunofluorescence were used to detect methylated NEAT1 in GC cells. Rescue assays were performed to define the relationship between NEAT1 and ALKBH5. NEAT1 is a potential binding lncRNA of ALKBH5. NEAT1 was overexpressed in GC cells and tissue. Additional experiments confirmed that knockdown of NEAT1 significantly repressed invasion and metastasis of GC cells. ALKBH5 affected the m(6)A level of NEAT1. The binding of ALKBH5 and NEAT1 influences the expression of EZH2 (a subunit of the polycomb repressive complex) and thus affects GC invasion and metastasis. Our findings indicate a novel mechanism by which ALKBH5 promotes GC invasion and metastasis by demethylating the lncRNA NEAT1. They may be potential therapeutic targets for GC.