1.Real-time PCR analysis of the apoptosis related genes in ATRA treated APL t(15;17) patients.
Hakan SAVLI ; Sema SIRMA ; Balint NAGY ; Melih AKTAN ; Guncag DINCOL ; Ugur OZBEK
Experimental & Molecular Medicine 2003;35(5):454-459
All-trans retinoic acid (ATRA) treatment of the acute promyelocytic leukemia (APL) have subsequently resulted in cell apoptosis, but the molecular mechanism of this effect remains elusive. In order to understand a possible involvement of genes regulating apoptotic signal pathways, expression levels of bcl2, bax, dapk1, myc, bad, wt1, and mcl genes were analyzed during ATRA treatment in five APL patients with t (15;17) using Real- time PCR (LightCycler). Two samples from each patient were compared to each other: primary diagnostic sample and a sample taken at remission. Effect of the ATRA treatment was demonstrated by the concomitant induction of cd14 and il1beta genes in four patients. Also other apoptosis related genes were found down-regulated in general but especially the down regulated levels of wt1 and bax attract attention. Result suggested that ATRA dependent apoptosis of APL was under the control of both internal and external pathways without relationships to the amount of the blast populations. Ratio of bcl2 to bax may be more important for this regulation than the ratio of bcl2 to bad. Either bcl2 family or less known apoptosis related genes as wt1 will still be required to further studies in this setting.
Apoptosis/drug effects/*genetics
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Chromosomes, Human, Pair 15/genetics
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Chromosomes, Human, Pair 17/genetics
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Gene Expression Profiling
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Gene Expression Regulation, Neoplastic/*drug effects
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HL-60 Cells
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Human
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Leukemia, Promyelocytic, Acute/*genetics
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Polymerase Chain Reaction/*methods
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Support, Non-U.S. Gov't
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Time Factors
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Translocation (Genetics)/*genetics
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Tretinoin/*pharmacology
2.Real-Time PCR analysis of af4 and dek genes expression in acute promyelocytic leukemiat (15;17)patients.
Hakan SAVLI ; Sema SIRMA ; Balint NAGY ; Melih AKTAN ; Guncag DINCOL ; Zafer SALCIOGLU ; Nazan SARPER ; Ugur OZBEK
Experimental & Molecular Medicine 2004;36(3):279-282
Among several newly identified oncogenes, dek and af4 are attractive targets for researchers interested with leukemia. In this study quantitative Real-Time RT-PCR technique was used to define alterations in expression of dek and af4 genes associated with acute promyelocytic leukaemia (APL) t (15; 17). RNA samples obtained from bone marrow aspirates of fourteen APL patients, cDNA portions were labelled with Syber Green 1 dye and LightCycler analysis have been performed. Expression changes in patients were found not significant in comparison to healthy donors for af4 (P=0.192) and dek (P= 0.0895). We suggest that af4 gene may have a role in leukomogenesis restricted to lymphoblastic lineage; also further studies must carry on with a larger series of patients in order to understand the relationship between the dek gene and APL. Our study was the first attempt for analysing dek and af4 genes in APL t (15; 17) patients by quantitative Real-Time RT-PCR. This rapid and sensitive method could be used to screen these genes in different types of leukaemia.
Adolescent
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Adult
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Child
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Child, Preschool
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Chromosomal Proteins, Non-Histone/*genetics/metabolism
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Chromosomes, Human, Pair 15
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Chromosomes, Human, Pair 17
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DNA-Binding Proteins/*genetics/metabolism
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Down-Regulation
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Female
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Gene Expression
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Humans
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Leukemia, Promyelocytic, Acute/*genetics/metabolism
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Male
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Middle Aged
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Nuclear Proteins/*genetics/metabolism
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Oncogene Proteins/*genetics/metabolism
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Polymerase Chain Reaction
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RNA, Messenger/analysis/metabolism
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Translocation, Genetic
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Up-Regulation