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Urea‐modulated UT‐B urea transporter internalization is clathrin‐ and caveolae‐dependent in infantile hemangioma‐derived vascular endothelial cells
Author(s) -
Xiao Li,
Liu Dakan,
Zuo Song,
Zhu Xiaoshuang,
Wang Yanlin,
Dong Changxian
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.27789
Subject(s) - caveolae , internalization , clathrin , urea , transporter , chemistry , endocytosis , microbiology and biotechnology , biochemistry , biology , signal transduction , receptor , gene
Abstract The aim of this study was to investigate the manner of urea‐modulated UT‐B urea transporter (UT) internalization in infantile hemangioma‐derived vascular endothelial cells (HemECs). The immunohistochemistry assay was performed to identify infancy hemangioma‐derived endothelial cell line (XPTS‐1) cells. Cell toxicity was detected with the 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyl tetrazolium bromide (MTT) assay. Quantitative real‐time polymerase chain reaction and Western blot analysis were measured to analyze the expression of UT‐B. UT‐B internalization was observed by confocal microscopy. The clathrin inhibitor chlorpromazine (CPZ) and caveolin endocytic disrupter methyl‐β‐cyclodextrin (MβCD) were used in XPTS‐1 cells transfected with UT‐B‐GFP to repress endocytosis. Urea‐promoted UT‐B expression in a concentration‐dependent manner in an infantile XPTS‐1 cell line. CPZ and MβCD significantly inhibited UT‐B protein internalization. The pretreatment of UT‐B‐GFP cells with adaptor protein2 (AP2)‐μ2‐siRNA and caveolin‐siRNA significantly inhibited UT‐B protein internalization. Our findings suggested that urea‐mediated UT‐B UT internalization is clathrin and caveolae dependent in infantile HemECs.