RESUMO
Aberrant expression of different SOX (SRY-related high mobility group (HMG) box) genes has been observed in number of tumors but, little is known about their expression patterns in hematological malignancies, especially in acute myeloid leukemia (AML). In this study we investigated SOX2, SOX3, SOX11, SOX14 and SOX18 gene expression in 50 de novo adult AML patients and correlated our findings with known clinical and molecular prognostic markers of the disease. We have found that these genes are overexpressed in 10-22% of patients and preliminary findings suggest that high expression level of these genes may have prognostic significance in AML patients. This is the first study focused on examining the expression level of SOX2, SOX3, SOX11, SOX14 and SOX18 genes in AML patients. Although this is a relatively limited study, initial findings indicate the need for further investigation of these genes, their potential roles in leukemia pathogenesis as well as prognosis in AML patients.
Assuntos
Perfilação da Expressão Gênica , Leucemia Mieloide Aguda/genética , Mutação , Fatores de Transcrição SOXB1/genética , Fatores de Transcrição SOXB2/genética , Fatores de Transcrição SOXC/genética , Fatores de Transcrição SOXF/genética , Adulto , Idoso , Biomarcadores Tumorais/genética , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Proteínas Nucleares/genética , Nucleofosmina , Prognóstico , Análise de Sobrevida , Adulto Jovem , Tirosina Quinase 3 Semelhante a fms/genéticaRESUMO
Sox3/SOX3 gene is implicated in the control of nervous system development and is considered to be one of the earliest neural markers. Expression of human SOX3 gene is modulated during the RA-induced neuronal differentiation cascade of NT2/D1 cells. Our present results demonstrate that the sequences responsible for RA-induced activation of SOX3 gene are localized within the 0.4 kb of its 5'-flanking region and implicate RXRalpha involvement in this regulation. The active RA/RXRalpha responsive region is pinned down to two regulatory elements. Only in the presence of both elements full RA/RXRalpha inducibility is achieved, suggesting they act synergistically. These elements comprise two unique G-rich boxes, separated by 49 bp, that could be considered as a novel, atypical RA-response element. Here, for the first time, we have demonstrated direct interaction of RXRalpha and SOX3 control elements. Furthermore, the functional in vivo analysis revealed that liganded RXRalpha is a potent activator of endogenous SOX3 protein expression. Since it is proven that Sox3 is critical determinant of neurogenesis our data may help in providing new insight into complex regulatory networks involved in retinoic acid induced neural differentiation of NT2/D1 cells.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas de Grupo de Alta Mobilidade/genética , Receptor X Retinoide alfa/metabolismo , Fatores de Transcrição/genética , Transcrição Gênica/efeitos dos fármacos , Tretinoína/farmacologia , Western Blotting , Células Cultivadas , Proteínas de Ligação a DNA/biossíntese , Desoxirribonuclease I/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Escherichia coli/metabolismo , Genes Reporter , Proteínas de Grupo de Alta Mobilidade/biossíntese , Humanos , Ligação Proteica , Proteínas Recombinantes/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição SOXB1 , Fatores de Transcrição/biossínteseRESUMO
We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHuIFN-beta1) was secreted from shake-flask-grown P. pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.u. (international units)/ml (6-12 mg/litre). An rHuIFN-beta1 with an N-terminal sequence identical with that of native HuIFN-beta1 was purified and the specific activity was determined (2-3 x 10(7) i.u./mg). It was found that the secreted recombinant protein was partially N-glycosylated.