Claudin-5 is involved in breast cancer cell motility through the N-WASP and ROCK signalling pathways.

Journal of Experimental & Clinical Cancer Research : CR
Astrid Escudero-EsparzaTracey A Martin

Abstract

Recent studies have shown dysregulation in TJ structure of several cancers including breast. Claudin-5 is a protein member of the TJ structure expressed in both endothelial and epithelial cells. This study examined the level of expression and distribution of Claudin-5 in human breast cancer tissues and the effect of knockdown and forced expression of Claudin-5 in the MDA-MB-231 breast cancer cell line. Immunohistochemistry and quantitative-PCR were used to analyse patient tissue samples. The Claudin-5 gene was cloned and overexpressed or knocked down using ribozyme technology in human breast cancer cells. Changes in function were assessed using in vitro assays for invasion, growth, adhesion, wounding, motility, transepithelial resistance and electric cell-substrate impedance sensing. Changes in cell behaviour were achieved through the use of Hepatocyte Growth factor (HGF) which we have shown to affect TJ function and expression of TJ proteins. In addition, an in vivo model was used for tumour growth assays. Results data was analyzed using a Students two sample t-test and by Two-way Anova test when the data was found to be normalized and have equal variances. In all cases 95% confidence intervals were used. Patients whose tumour...Continue Reading

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Citations

Mar 26, 2013·BMC Cell Biology·Patricia G WebbHeidi K Baumgartner
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Methods Mentioned

BETA
transfect
PCR
co-immunoprecipitation
Protein Assay
motility assay
MDA
immunoprecipitation
motility

Software Mentioned

SigmaPlot
Beacon Designer
ECIS RbA
RNA mFold
SPSS

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