Lai Deng-Pan, Tan Sheng, Kang Ya-Ni, Wu Jun, Ooi Hong-Sain, Chen Jian, Shen Ting-Ting, Qi Ying, Zhang Xiaoli, Guo Yan, Zhu Tao, Liu Bingya, Shao Zhifeng, Zhao Xiaodong
State Key Laboratory for Oncogenes and Related Genes, Bio-ID Center, School of Biomedical Engineering, Ruijin Hospital.
Laboratory of Molecular Tumor Pathology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China.
Hum Mol Genet. 2015 Jun 15;24(12):3410-7. doi: 10.1093/hmg/ddv089. Epub 2015 Mar 10.
Alternative polyadenylation (APA) is an important post-transcriptional modification implicated in many diseases, including cancer. Although extensively characterized, the functional consequence of APA modulation on tumorigenesis remains elusive. Here, we developed a deep sequencing-based approach that specifically profiles 3' termini of polyadenylated RNAs (herein termed 3T-seq) and analyzed APA events in two gastric cancer cell lines and one non-transformed counterpart. Overall, we identified >28 000 poly(A) sites, 70% of which are potentially novel. Further, we observed widespread APA-mediated 3' UTR shortening of 513 genes (false discovery rate < 0.05) across gastric cancer genome. We characterized one of these genes, NET1, in detail and found that the shortening of NET1 3' UTR significantly enhances transcriptional activity. Moreover, the NET1 isoform with short 3' UTR promotes cellular migration and invasion in vitro. Collectively, our work provides an effective approach for genome-wide APA site profiling and reveals a link between APA modulation and gastric cancer metastasis.
可变聚腺苷酸化（APA）是一种重要的转录后修饰，与包括癌症在内的多种疾病有关。尽管已被广泛研究，但APA调控对肿瘤发生的功能后果仍不清楚。在此，我们开发了一种基于深度测序的方法，专门分析聚腺苷酸化RNA的3'末端（以下称为3T-seq），并分析了两种胃癌细胞系和一种未转化的对照细胞系中的APA事件。总体而言，我们鉴定出超过28000个聚腺苷酸位点，其中70%可能是新发现的。此外，我们在胃癌基因组中观察到513个基因存在广泛的APA介导的3'UTR缩短（错误发现率<0.05）。我们详细研究了其中一个基因NET1，发现NET1 3'UTR的缩短显著增强了转录活性。此外，具有短3'UTR的NET1异构体在体外促进细胞迁移和侵袭。总的来说，我们的工作为全基因组APA位点分析提供了一种有效方法，并揭示了APA调控与胃癌转移之间的联系。
