RNA interference for HIF-1α inhibits its downstream signalling and affects cellular proliferation

被引：88

作者：

Hänze, J ^{[1
]}

Eul, BG ^{[1
]}

Savai, R ^{[1
]}

Krick, S ^{[1
]}

Goyal, P ^{[1
]}

Grimminger, F ^{[1
]}

Seeger, W ^{[1
]}

Rose, F ^{[1
]}

机构：

[1] Univ Giessen, Sch Med, Dept Internal Med 2, D-35392 Giessen, Germany

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2003年 / 312卷 / 03期

关键词：

HIF-1; RNA interference; siRNA; cell cycle; proliferation; hypoxia;

D O I：

10.1016/j.bbrc.2003.10.153

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Transcription factor HIF-1 is a key determinant of oxygen-dependent gene regulation. Suppression of HIF-1alpha is important for exploring HIF-1-dependent processes and for interfering with hypoxia-induced pathophysiological events. This study applied RNA-interference targeting HIF-1alpha to the human lung A549 cell line. Transfection of HIF-1alpha-siRNA reduced HIF-1alpha synthesis as measured on mRNA and protein level by realtime RT-PCR, Western blot, and immuncytochemistry. A time kinetic for hypoxic stabilization of HIF-1alpha protein and its inhibition by HIF-1alpha-siRNA is included. Hypoxic induction of HIF-1-controlled target genes as heme oxyaenase 1 (HO-1), phosphoglycerate kinase (PGK), and vascular endothelial growth factor (VEGF) was markedly attenuated by HIF-1alpha-siRNA treatment. Correspondingly, gene activation via hypoxia-responsive-element, as shown by reporter gene assay, was inhibited by HIF-1alpha-siRNA. Moreover, this approach was found to suppress the shift from from S-phase to G(1)-phase observed in A549 cells in response to hypoxia, supporting a role of HIF-1alpha in oxygen-dependent cell cycle regulation. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：571 / 577

页数：7

共 33 条

[1] A conserved family of prolyl-4-hydroxylases that modify HIF [J].