Hypoxic induction of hypoxia-inducible factor-1alpha and oxygen-regulated gene expression in mitochondrial DNA-depleted HeLa cells.
Hypoxia increases the accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) and induces transcription of a variety of genes including vascular endothelial growth factor (VEGF) gene through oxygen sensing mechanisms. In one model, mitochondria-derived reactive oxygen species (ROS) are suggested to be involved in oxygen sensing. Here, we found that hypoxia increased ROS generation and VEGF gene expression in HeLa cells. To further investigate the role of ROS in oxygen sensing, we compared the hypoxic response between rho(0) HeLa cells lacking mitochondrial DNA (EB8) and rho(0) HeLa cells containing mitochondrial DNA from wild-type HeLa cells (HeEB1). The results showed that, although hypoxia markedly increased ROS generation in HeEB1 cells but not in EB8 cells, EB8 cells showed essentially a normal response to hypoxia, as assessed by VEGF gene promoter activity, HIF-1alpha accumulation, and HIF-1 target gene expressions. These results indicate that mitochondria-derived ROS generated in response to hypoxia are not necessary for oxygen sensing in HeLa cells.
Enomoto
Noriko
N
Division of Chemotherapy, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuoh-ku, Japan.
Koshikawa
Nobuko
N
Gassmann
Max
M
Hayashi
Jun-Ichi
J
Takenaga
Keizo
K
D015687:Cell Hypoxia; D004272:DNA, Mitochondrial / Q000378:metabolism*; D016228:Endothelial Growth Factors / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008222:Lymphokines / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D004272:DNA, Mitochondrial; D016228:Endothelial Growth Factors; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D017382:Reactive Oxygen Species; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D010100:Oxygen