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Treatment of the C6 rat glioma cell variant, STI cells, with glucocorticoids, leads to a complete reversion of their transfonned phenotype and loss of tumorigenic potential. ln this work, two subtracted and equalized cDNA libraries were constructed, based on suppressive PCR, in order to identify glucocorticoidregulated genes related to the ST 1 phenotypic reversion. Analysis of both subtracted cDNA libraries revealed 7 differentially expressed genes, some of which are well known negative growth regulators: thrombospondin-1, cyclin G, tyrosine phosphatase CL 100 and NRP/B. ln addition, a cDNA clone similar to TN-X, but exibiting a different transcript size was also isolated. lsolation and characterization of a full-length cDNA corresponding to this transcript, revealed that it is composed of fibronectin type III repeats and one single fibrinogen domain. Glucocorticoid induction of this transcript may suggest a different role of TN-X variants in controling cell proliferation. Expression analysis of the C6-derived glucocorticoid-resistant, P7 variant, and of the human glioblastoma U87 cells, revealed very low or undectable leveis of these transcripts, further suggesting that their expression is related to the STI cells' phenotypic reversion. Prevalence of rare and differentially expressed transcripts m these libraries suggests that they are likely to be useful in automated systems (DNA microarrays) to rapidly validate differentially expressed clones and to reveal a panei of candidate genes that mediate the anti-proliferative and anti-tumor effects of glucocorticoids.

Isolamento, identificação e caracterização de seqüências expressas de DNA (ESTs) de genes regulados por glicocorticóides na reversão fenotípica tumoral-normal de células ST1 de glioma de rato