Antisense-thioredoxin inhibits angiogenesis via pVHL-mediated hypoxia-inducible factor-1alpha degradation.
Thioredoxin (Trx), a small redox protein, is overexpressed in a number of tumors, however, its roles in tumor cells were not defined well. To investigate the effect of Trx, we transfected Trx or antisense Trx vectors into HT1080 cells. Trx-overexpressed HT1080 cells induced migration of endothelial cells through Flt-1 vascular endothelial growth factor (VEGF) receptor type-1. However, the migration was reduced by overexpression of antisense-Trx. To understand the relationship between Trx and hypoxia-induced angiogenesis, we observed the expression level of VEGF and hypoxia-inducible factor-1alpha (HIF-1alpha) after transfection of Trx. Overexpression of Trx also caused a significant increase of VEGF in protein and RNA levels under normoxic and hypoxic conditions. Moreover, transfection of Trx caused a dramatic increase in HIF-1alpha protein level under hypoxic condition. However, transfection of antisense-Trx markedly decreased HIF-1alpha and VEGF expression compared with controls. In addition, HIF-1alpha was not translocated from cytoplasm to nucleus in antisense-Trx overexpressing HT1080 cells. Further, we could detect the association of HIF-1alpha and pVHL in antisense-Trx transfectant HT1080 cells. Taken together, we suggest that Trx plays an important role in angiogenesis and, therefore, antisense-Trx might be applicable to the inhibition of tumor angiogenesis through the induction of pVHL-mediated HIF-1alpha degradation.
Kim
Woo Jean
WJ
Department of Molecular Biology, Pusan National University, Busan 609-735, Korea.
Cho
Heeyeong
H
Lee
Sae-Won
SW
Kim
Yung-Jin
YJ
Kim
Kyu-Won
KW
D015152:Blotting, Northern; D015153:Blotting, Western; D015687:Cell Hypoxia; D002465:Cell Movement; D015536:Down-Regulation; D042783:Endothelial Cells; D016147:Genes, Tumor Suppressor; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009369:Neoplasms / Q000098:blood supply; D009389:Neovascularization, Pathologic; D016376:Oligonucleotides, Antisense; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013879:Thioredoxins / Q000235:genetics* / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism / Q000494:pharmacology*; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000494:pharmacology*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000494:pharmacology*; D014470:Umbilical Cord / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*; D051794:Von Hippel-Lindau Tumor Suppressor Protein
C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016376:Oligonucleotides, Antisense; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D042461:Vascular Endothelial Growth Factor A; D013879:Thioredoxins; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human