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Hypoxia-induced production of peptidylarginine deiminases and citrullinated proteins in malignant glioma cells.

Abstract：

BACKGROUND:Recently, it has been reported that hypoxia highly enhances expression of peptidylarginine deiminase (PAD) 4 and production of citrullinated proteins in some tumor cells. However, little is known about malignant gliomas on this issue. Therefore, we here investigated whether expression of PADs was induced by hypoxia and whether PADs citrullinated intracellular proteins if induced using U-251 MG cells of a human malignant glioma cell line. METHODS:Expression of PADs in U-251 MG cells, cultured under hypoxia or normoxia for 24 h, was investigated by quantitative polymerase chain reaction (qPCR). Citrullination of proteins in the cells and the cell lysates incubated for 48 h with or without Ca2+ was detected by western blotting. Citrullinated proteins were identified by mass spectrometry. RESULTS:The mRNA levels of PAD1, 2, 3, and 4 were up-regulated by hypoxia in a hypoxia-inducible factor-1-dependent manner in U-251 MG cells. In spite of the increased expression, intracellular proteins were not citrullinated. However, the induced PADs citrullinated U-251 MG cell-derived proteins when the cells were lysed. Multiple proteins citrullinated by hypoxia-induced PADs were identified. In addition, the extracellular domain of vascular endothelial growth factor receptor 2 was citrullinated by human PAD2 in vitro. CONCLUSION:Our data may contribute to understanding of pathophysiology of malignant gliomas from the aspects of protein citrullination.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Sase T,Arito M,Onodera H,Omoteyama K,Kurokawa MS,Kagami Y,Ishigami A,Tanaka Y,Kato T

doi

10.1016/j.bbrc.2016.10.154

subject

Has Abstract

pub_date

2017-01-01 00:00:00

pages

50-56

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(16)31792-2

journal_volume

482

pub_type

杂志文章

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