RREB1-induced upregulation of the lncRNA AGAP2-AS1 regulates the proliferation and migration of pancreatic cancer partly through suppressing ANKRD1 and ANGPTL4
RREB1-induced upregulation of the lncRNA AGAP2-AS1 regulates the proliferation and migration of pancreatic cancer partly through suppressing ANKRD1 and ANGPTL4
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RREB1诱导的lncRNA AGAP2-AS1上调部分通过抑制ANKRD1和ANGPTL4调节胰腺癌的增殖和迁移
DOI：
10.1038/s41419-019-1384-9
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发表时间：
2019-02-27
影响因子：
9
通讯作者：
Zhou, Yan
中科院分区：
文献类型：
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作者：
Hui, Bingqing;Ji, Hao;Zhou, Yan
据报道，长非编码RNA(LncRNAs)与多种人类疾病有关，包括癌症。然而，它们的作用机制尚未完全阐明。我们通过分析已发表的微阵列数据，研究了可能与胰腺癌(PC)相关的LncRNA变化，并确定AGAP2-AS1是PC组织中相对高表达的LncRNA。采用定量RT-PCR方法检测AGAP2-AS1的表达水平。用四甲基偶氮唑盐比色法、集落形成实验和EDU实验检测细胞的增殖能力。采用流式细胞仪和原位末端标记法研究AGAP2-AS1在细胞周期和细胞凋亡中的调控作用。通过Transwell实验研究细胞侵袭和转移的变化，并建立裸鼠模型以评估AGAP2-AS1对体内肿瘤形成的影响。RNA测序检测AGAP2-AS1相关通路。亚细胞分级和FISH检测AGAP2-AS1在PC细胞中的分布，RIP和CHIP检测AGAP2-AS1对潜在靶基因调控的分子机制。在PC患者中，AGAP2-AS1的表达增加与肿瘤大小和病理分期有关。RREB1可激活PC细胞中AGAP2-AS1的转录。AGAP2-AS1在体外影响PC细胞的增殖、凋亡、周期停滞、侵袭和转移，而AGAP2-AS1在体内调节PC的增殖。此外，AGAP2-AS1通过募集Zust Homolog 2(EZH2)抑制ANKRD1和ANGPTL4的表达，从而促进PC的增殖和转移。综上所述，我们的数据表明，RREB1诱导的AGAP2-AS1上调部分是通过招募EZH2抑制ANKRD1和ANGPTL4，从而调节PC细胞的增殖和迁移。AGAP2-AS1是未来PC诊断和治疗的潜在靶点。
Long noncoding RNAs (lncRNAs) have been reported to be involved in a variety of human diseases, including cancers. However, their mechanisms have not yet been fully elucidated. We investigated lncRNA changes that may be associated with pancreatic cancer (PC) by analyzing published microarray data, and identified AGAP2-AS1 as a relatively overexpressed lncRNA in PC tissues. qRT-PCR assays were performed to examine expression levels of AGAP2-AS1. MTT assays, colony formation assays, and EdU assays were used to determine the proliferative capacity of cells. Flow cytometry and TUNEL assays were used to study the regulation of AGAP2-AS1 in the cell cycle and apoptosis. Transwell experiments were used to study changes in cell invasion and metastasis, and a nude mouse model was established to assess the effects of AGAP2-AS1 on tumorigenesis in vivo. RNA sequencing was performed to probe AGAP2-AS1-related pathways. Subcellular fractionation and FISH assays were used to determine the distribution of AGAP2-AS1 in PC cells, and RIP and ChIP were used to determine the molecular mechanism of AGAP2-AS1-mediated regulation of potential target genes. Increased expression of AGAP2-AS1 was associated with tumor size and pathological stage progression in patients with PC. RREB1 was found to activate transcription of AGAP2-AS1 in PC cells. AGAP2-AS1 affected proliferation, apoptosis, cycle arrest, invasion, and metastasis of PC cells in vitro, and AGAP2-AS1 regulated PC proliferation in vivo. Furthermore, AGAP2-AS1 epigenetically inhibited the expression of ANKRD1 and ANGPTL4 by recruiting zeste homolog 2 (EZH2), thereby promoting PC proliferation and metastasis. In summary, our data show that RREB1-induced upregulation of AGAP2-AS1 regulates cell proliferation and migration in PC partly through suppressing ANKRD1 and ANGPTL4 by recruiting EZH2. AGAP2-AS1 represents a potential target for the diagnosis and treatment of PC in the future.