Effect of human S100A13 gene silencing on FGF-1 transportation in human endothelial cells.
OBJECTIVE
The S100 protein is part of a Ca(2+) binding protein superfamily that contains an EF hand domain, which is involved in the onset and progression of many human diseases, especially the proliferation and metastasis of tumors. S100A13, a new member of the S100 protein family, is a requisite component of the fibroblast growth factor-1 (FGF-1) protein release complex, and is involved in human tumorigenesis by interacting with FGF-1 and interleukin-1. In this study, experiments were designed to determine the direct role of S100A13 in FGF-1 protein release and transportation.


METHODS
We successfully constructed the lentiviral vectors containing shRNA targeting the human S100A13 gene. Human umbilical vein endothelial cells (HUVECs) were transfected with lentiviral RNAi vectors for S100A13. Then immunofluorescence staining, real-time quantitative polymerase chain reaction and Western blotting were used to detect the inhibition efficiency of the vectors and to monitor the release and transportation of FGF-1 protein.


RESULTS
Lentiviral RNAi vectors induced suppression efficiency of S100A13 gene by 90% in HUVECs. FGF-1 protein was found to be transported from the cytoplasm to the cell membrane, and then released from cells when HUVECs were deprived of serum. The release of FGF-1 protein was blocked by the downregulation of S100A13, but the transportation was not affected, suggesting that S100A13 is a key cargo protein for FGF-1 release.


CONCLUSIONS
S100A13 promotes the release of FGF-1 protein, but does not affect the transportation of FGF-1 protein in HUVECs.
Cao
Renxian
R
Department of Pathophysiology, University of South China, Hengyang, China. caorenxian@hotmail.com
Yan
Bin
B
Yang
Huiling
H
Zu
Xuyu
X
Wen
Gebo
G
Zhong
Jing
J
D015153:Blotting, Western; D015536:Down-Regulation / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism*; D016220:Fibroblast Growth Factor 1 / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D020868:Gene Silencing; D006801:Humans; D017958:Lentiviruses, Primate; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger; D009418:S100 Proteins / Q000235:genetics / Q000502:physiology*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism
D012333:RNA, Messenger; D009418:S100 Proteins; C511523:S100A13 protein, human; D016220:Fibroblast Growth Factor 1