Overexpression of S100A4 in human cancer cell lines resistant to methotrexate

BMC Cancer

Research Article

Véronique Noé¹  Isabel Rico¹  M Cristina de Almagro¹  Carlos J Ciudad¹  Nuria Mencía¹  Elisabet Selga¹  Xenia Villalobos¹  Sara Ramirez¹  Jaume Adan²  Jose L Hernández² 

[1] Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, Diagonal Avenue 643, Barcelona, Spain;Leitat Technological Center (Biomed Division), Helix building PCB, Barcelona, Spain;

关键词: HT29 Cell;    Resistant Cell;    Resistant Cell Line;    S100A4 Expression;    S100A4 mRNA;   

DOI  :  10.1186/1471-2407-10-250

 received in 2009-10-21, accepted in 2010-06-01,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundMethotrexate is a chemotherapeutic drug that is used in therapy of a wide variety of cancers. The efficiency of treatment with this drug is compromised by the appearance of resistance. Combination treatments of MTX with other drugs that could modulate the expression of genes involved in MTX resistance would be an adequate strategy to prevent the development of this resistance.MethodsThe differential expression pattern between sensitive and MTX-resistant cells was determined by whole human genome microarrays and analyzed with the GeneSpring GX software package. A global comparison of all the studied cell lines was performed in order to find out differentially expressed genes in the majority of the MTX-resistant cells. S100A4 mRNA and protein levels were determined by RT-Real-Time PCR and Western blot, respectively. Functional validations of S100A4 were performed either by transfection of an expression vector for S100A4 or a siRNA against S100A4. Transfection of an expression vector encoding for β-catenin was used to inquire for the possible transcriptional regulation of S100A4 through the Wnt pathway.ResultsS100A4 is overexpressed in five out of the seven MTX-resistant cell lines studied. Ectopic overexpression of this gene in HT29 sensitive cells augmented both the intracellular and extracellular S100A4 protein levels and caused desensitization toward MTX. siRNA against S100A4 decreased the levels of this protein and caused a chemosensitization in combined treatments with MTX. β-catenin overexpression experiments support a possible involvement of the Wnt signaling pathway in S100A4 transcriptional regulation in HT29 cells.ConclusionsS100A4 is overexpressed in many MTX-resistant cells. S100A4 overexpression decreases the sensitivity of HT29 colon cancer human cells to MTX, whereas its knockdown causes chemosensitization toward MTX. Both approaches highlight a role for S100A4 in MTX resistance.

【 授权许可】

Unknown   
© Mencía et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.