Identification of okadaic-acid-induced genes by mRNA differential display in glioma cells

To identify novel genes associated with apoptosis in glioma cells, we treat ed T98G glioma cells with okadaic acid (OA). Differential display using 15 random primers was performed on RNA extracted from these cells. Upregulated bands were excised from polyacrylamide gels and cloned. Northern blots wer e used to confirm RNA expression in T98G cells, 18 RNA fragments correspond ing to the untranslated region of genes were identified and sequenced. Thre e unknown gene fragments were used to screen a fetal brain cDNA library res ulting in three complete cDNA sequences. The three sequences corresponded t o a human gene homologous to the yeast translation initiation factor Sui-l, a cAMP-regulated phosphoprotein, ARPP-16/19, and a novel gene designated O 48, Transcription of Sui-l increased in response to all stress factors test ed, whereas ARPP only responded to OA, 2-kb and 4-kb O48 RNA species were i dentified. OA and stress factors increased 2-kb expression while K252a (pro tein kinase inhibitor) increased 4-kb expression, Differential display is e ffective for identifying genes associated with apoptosis, Novel genes may b e identified by further analysis of the gene fragments identified in this s tudy. The function of O48 is unknown. Copyright (C) 2000 National Science C ouncil, ROC and S. Karger AG, Basel.