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Molt-4 leukemia cells undergo p53-dependent apoptosis accompanied by accumulation of  de novo  ceramide after 14 hours of γ-irradiation. In order to identify the potential mediators involved in ceramide accumulation and the cell death response, differentially expressed genes were identified by Affymetrix Microarray Analysis. Molt-4-LXSN cells, expressing wild type p53, and p53-deficient Molt-4-E6 cells were irradiated and harvested at 3 and 8 hours post-irradiation. Human genome U133 plus 2.0 array containing >47,000 transcripts was used for gene expression profiling. From over 10,000 probes, 281 and 12 probes were differentially expressed in Molt-4-LXSN and Molt-4-E6 cells, respectively. Data analysis revealed 63 (upregulated) and 20 (downregulated) genes (>2 fold) in Molt-4-LXSN at 3 hours and 140 (upregulated) and 21 (downregulated) at 8 hours post-irradiation. In Molt-4-E6 cells, 5 (upregulated) genes each were found at 3 hours and 8 hours, respectively. In Molt-4-LXSN cells, a significant fraction of the genes with altered expression at 3 hours were found to be involved in apoptosis signaling pathway ( BCL2L11 ), p53 pathway ( PMAIP1 ,  CDKN1A  and  FAS ) and oxidative stress response ( FDXR ,  CROT  and  JUN ). Similarly, at 8 hours the genes with altered expression were involved in the apoptosis signaling pathway ( BAX ,  BIK  and  JUN ), p53 pathway ( BAX ,  CDKN1A  and  FAS ), oxidative stress response ( FDXR  and  CROT ) and p53 pathway feedback loops 2 ( MDM2  and  CDKN1A ). A global molecular and biological interaction map analysis showed an association of these altered genes with apoptosis, senescence, DNA damage, oxidative stress, cell cycle arrest and caspase activation. In a targeted study, activation of apoptosis correlated with changes in gene expression of some of the above genes and revealed sequential activation of both intrinsic and extrinsic apoptotic pathways that precede ceramide accumulation and subsequent execution of apoptosis. One or more of these altered genes may be involved in p53-dependent ceramide accumulation.

Genomic alterations during p53-dependent apoptosis induced by γ-irradiation of Molt-4 leukemia cells