CML transcriptome analysis using ORESTES (Open Reading Frame Expression Sequence Tags) / Avaliação do transcriptoma da leucemia mieloide cronica por ORESTES (Open Reading Frame Expression Sequence Tags)

AUTOR(ES)
DATA DE PUBLICAÇÃO

2003

RESUMO

The complete collection of transcripts generated from the human genome cannot be predicted from the genome sequence, but should be directly determined for each tissue, due to variations of gene expression in different tissues and disease states, and because genes can encode multiple transcripts derived from alternate splicing and polyadenylation sites. As part of larger project that produced up to 1.2 million ORESTES (open reading frames expressed sequence tags) from different cancer tissues, we constructed a set of cDNAs obtained from bone marrow cells of patients with CML, that represented partial expressed gene sequences that are biased toward the central coding regions of the resulting transcripts (Dias-Neto E et al, Proc Nat Acad Sci USA 97:3491, 2000). The 27,361 ESTs were assembled into contigs using the Phrap package producing 2933 clusters containing 2 to 210 ESTs (leaving 11568 isolated sequences). Functional categories were attributed to the contigs on the basis of the annotation of the matched sequences in nr, into 5 major functional categories and 22 subcategories (cell communication (19%); cell growth, development and m contigs formed by the larger number of EST in bone marrow cells was co mpared with a control bone marrow sample obtained from a non-neoplastic hematologic condition (hereditary hemolytic anemia) and evaluated with SAGE (serial analysis of gene expression). Ten out of 100 most representative contigs had their expression data evaluated further by realtime quantitative PCR with Sybr Green I reagent, confirming the ORESTES data. Seven out of the 10 genes were more than 10-fold upregulated (CAMP, ABR, DEFA3, CSF3R, MLL2, S100A8 e MS4A3) and two additional genes were 50-fold or higher upregulated in CML (DEFA3 e ABR). The wealthy of information provided by this approach demonstrates its usefulness for the analysis of gene expression in specific hematopoietic tissues and diseases

ASSUNTO(S)

gene expression chronic leukemia reação em cadeia da polimerase via transcriptase reversa expressão genica myeloid leucemia mieloide cronica reverse transcriptase polymerase chain reaction

ACESSO AO ARTIGO

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