Proteomic analysis of gene expression following hypoxia and reoxygenation reveals proteins involved in the recovery from endoplasmic reticulum and oxidative stress.
OBJECTIVE
Human tumors are characterized by large variations in oxygen concentration and hypoxic tumors are associated with poor prognosis. In addition, tumors are subjected to periodic changes in oxygenation characterized by hypoxia followed by reoxygenation. Cellular adaptation to hypoxia is well documented, nevertheless little is known about adaptive mechanisms to reoxygenation. Here, we investigate the changes in protein expression during reoxygenation using proteomics.


METHODS
HeLa cervix carcinoma cells were exposed to 4h of hypoxia (<0.01% O(2)) followed by 1h of reoxygenation. The cellular proteome was examined using 2D gel electrophoresis coupled with mass spectrometry. Validation and investigation of the underlying basis for induced protein expression was investigated using Western blot analysis and quantitative RT-PCR.


RESULTS
We identified proteins involved in several cellular processes that are responsible for regulating RNA metabolism, protein synthesis and degradation, including ribosomal protein P0, VCP/p97 and FUSE binding protein 2.


CONCLUSIONS
Our results suggest that these newly identified proteins function in pathways that may assist in the recovery of ER stress and protein synthesis during reoxygenation. These proteins may thus be important determinants of the behaviour and survival of tumor cells to transient hypoxic exposures.
Magagnin
Michaël G
MG
Department of Radiation Oncology (Maastro Lab), GROW Research Institute, Maastricht University, The Netherlands.
Sergeant
Kjell
K
van den Beucken
Twan
T
Rouschop
Kasper M A
KM
Jutten
Barry
B
Seigneuric
Renaud
R
Lambin
Philippe
P
Devreese
Bart
B
Koritzinsky
Marianne
M
Wouters
Bradly G
BG
D015153:Blotting, Western; D015687:Cell Hypoxia; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006367:HeLa Cells; D006801:Humans; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000235:genetics*; D010100:Oxygen / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D040901:Proteomics
D010100:Oxygen