Analysis of HIF dependent transcriptional regulation resolved by transcriptional start site, using 5' end-Seq technology

Changes in mRNA abundance upon HIF activation and/or hypoxia in VHL-defective kidney cancer cell were analysed. Sequencing of the 5' ends of mRNAs (5' end-Seq) was applied to VHL-defective kidney cancer cell lines with or without reintroduction of VHL and/or with hypoxic incubation (1% O2 for 24h) to analyse the abundance of mRNAs resolved by their transcription start sites. In addition, the contribution of HIF pathway was confirmed using 786-O cells in which HIF1B was inactivated.

本研究分析了VHL（von Hippel-Lindau）缺陷型肾癌细胞在缺氧诱导因子（Hypoxia-Inducible Factor, HIF）激活及/或缺氧条件下的mRNA丰度变化。我们采用mRNA 5'端测序（5' end-Seq）技术，对分别导入或未导入VHL基因、并经/未经缺氧孵育（1% O₂培养24小时）的VHL缺陷型肾癌细胞系开展检测，以解析不同转录起始位点对应的mRNA丰度。此外，本研究利用HIF1B（Hypoxia-Inducible Factor 1 Beta, HIF1B）基因失活的786-O细胞，验证了HIF信号通路的相关调控作用。