Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.intimp.2007.03.002
Title: Gab2 antisense oligonucleotide blocks rat basophilic leukemic cell functions
Authors: Chan, J.H.P.
Liao, W. 
Wong, W.S.F. 
Lau, H.Y.A.
Keywords: Adhesion
Akt
FcεRI
p38 MAPK
RBL-2H3
Issue Date: 2007
Source: Chan, J.H.P., Liao, W., Wong, W.S.F., Lau, H.Y.A. (2007). Gab2 antisense oligonucleotide blocks rat basophilic leukemic cell functions. International Immunopharmacology 7 (7) : 937-944. ScholarBank@NUS Repository. https://doi.org/10.1016/j.intimp.2007.03.002
Abstract: Adapter molecule Grb2-associated binder-like protein 2 (Gab2) plays a critical role in FcεRI-induced mast cell degranulation and activation. The present study aimed to investigate the pharmacological effects of an antisense oligonucleotide (ASO) targeted at Gab2 on the immune responses of rat basophilic leukemic (RBL)-2H3 cells. Gab2 ASOs were rationally designed and transfected into RBL-2H3 cells. Gab2 mRNA and protein knockdown was confirmed by real-time RT-PCR and immunoblotting, respectively. Effects of Gab2 ASO on FcεRI-induced release of histamine and β-hexosaminidase was measured by EIA and an enzymatic assay, respectively; signaling events by immunoblotting; and cytokine mRNA expression by RT-PCR. Effects of Gab2 ASO on cell adhesion and migration were performed on fibronectin-coated 96-well plate and transwells cell culture chambers, respectively. We have characterized a phosphorothioate-modified ASO targeted at Gab2 mRNA that was able to knockdown Gab2 mRNA and protein in RBL-2H3 cells. Gab2 ASO significantly blocked IgE-mediated mast cell release of β-hexosaminidase and histamine; phosphorylation of Akt, p38 mitogen-activated protein kinase and PKCδ; and up-regulation of cytokine mRNA levels (e.g. IL-4, -6, -9 and -13, and TNF-α). In addition, Gab2 ASO markedly prevented mast cell adhesion to fibronectin-coated plates and restrained random migration of RBL-2H3 cells in cell culture chambers. Our findings show that Gab2 knockdown in RBL-2H3 cells by ASO strategy can suppress many aspects of the mast cell functions and, therefore, a selective Gab2 ASO may have therapeutic potential for mast cell-dependent allergic disorders. © 2007 Elsevier B.V. All rights reserved.
Source Title: International Immunopharmacology
URI: http://scholarbank.nus.edu.sg/handle/10635/27304
ISSN: 15675769
DOI: 10.1016/j.intimp.2007.03.002
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

12
checked on Dec 6, 2017

WEB OF SCIENCETM
Citations

12
checked on Nov 21, 2017

Page view(s)

195
checked on Dec 10, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.