Control of hypoxia-induced tumor cell adhesion by cytophilic human catalase.
Hypoxia-induced reactive oxygen species (ROS)-mediated expression of a variety of genes in endothelial cells has been suggested to be involved in abnormal cell adhesion. To prevent this by accelerated binding of catalase to endothelial cells, human catalase (hCAT), an enzyme catalyzing the decomposition of hydrogen peroxide, was fused with three repeats of arginine-glycine-aspartic acid peptide or nona arginine peptide at the C-terminal to obtain hCAT-(RGD)3 and hCAT-R9, respectively. Human CAT and its derivatives were expressed in yeast Pichia pastoris and purified. The specific activity and secondary structure of hCAT-(RGD)3 and hCAT-R9 were close to those of hCAT, but these derivatives showed higher binding to the mouse aortic vascular endothelial cell line MAEC than hCAT, indicating that they are cytophilic derivatives. Hypoxic treatment of MAEC increased the intracellular ROS level, the binding of mouse melanoma cells, and the activity of transcription factors, hypoxia inducible factor-1 and nuclear factor-kappaB. hCAT-(RGD)3 or hCAT-R9 efficiently inhibited these changes compared with hCAT. These results indicate that cytophilic hCAT-(RGD)3 and hCAT-R9 are effective in inhibiting hypoxia-induced tumor cell adhesion to endothelial cells.
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Science, Kyoto University, Sakyo-ku, Kyoto 606-8501, Sakyo-ku, Japan.
Tomoya
T
Yata
Makiya
M
Nishikawa
Chika
C
Nishizaki
Masahide
M
Oku
Hiroya
H
Yurimoto
Yasuyoshi
Y
Sakai
Yoshinobu
Y
Takakura
D000818:Animals; D002374:Catalase / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D002415:Cats; D002448:Cell Adhesion / Q000187:drug effects; D015687:Cell Hypoxia; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D051793:Hypoxia-Inducible Factor 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D009369:Neoplasms / Q000201:enzymology* / Q000473:pathology; D009842:Oligopeptides / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*
D051793:Hypoxia-Inducible Factor 1; D016328:NF-kappa B; D009842:Oligopeptides; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; C491427:nonaarginine; C047981:arginyl-glycyl-aspartic acid; D006861:Hydrogen Peroxide; D002374:Catalase