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Modulation of IMPDH2, survivin, topoisomerase I and vimentin increases sensitivity to methotrexate in HT29 human colon cancer cells
Author(s) -
Peñuelas Silvia,
Noé Véronique,
Ciudad Carlos J.
Publication year - 2005
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2004.04504.x
Subject(s) - survivin , vimentin , biology , microbiology and biotechnology , cancer research , gene , topoisomerase , methotrexate , chemistry , biochemistry , enzyme , immunology , immunohistochemistry
We determined differentially expressed genes in HT29 human colon cancer cells, both after short treatment with methotrexate (MTX) and after the resistance to MTX had been established. Screening was performed using Atlas Human Cancer 1.2K cDNA arrays. The analysis was carried out using Atlas image 2.01 and genespring 6.1 software. Among the differentially expressed genes we chose for further validation were inosine monophosphate dehydrogenase type II ( IMPDH2 ), inosine monophosphate cyclohydrolase and survivin as up‐regulated genes, and topoisomerase I ( TOP1 ) and vimentin as down‐regulated genes. Changes in mRNA levels were validated by quantitative RT‐PCR. Additionally, functional analyses were performed inhibiting the products of the selected genes or altering their expression to test if these genes could serve as targets to modify MTX cytotoxicity. Inhibition of IMPDH or TOP1 activity, antisense treatment against survivin, or overexpression of vimentin, sensitized resistant HT29 cells to MTX. Therefore, these proteins could constitute targets to develop modulators in MTX chemotherapy.