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Real-time quantitative PCR analysis of human leukemic B-cells.

ABSTRACT: Fresh blood samples were collected from 10 CLL (chronic lymphocytic leukemia) previously untreated patients before and after 2 weeks of the first cycle of CCR (cladribine, cyclophosphamide, rituximab) treatment. Peripheral blood mononuclear cells (PBMNCs) were separated from EDTA blood by layering on the Histopaque 1077 and centrifuging on a density gradient. Mean B-cell CD19+ purity was ≥95% as measured by flow cytometry (FC). Microarray analysis was performed using 0.5 microgram of RNA transcribed to cDNA. Prepared cDNA samples were subjected to RT-PCR in duplicate in the TaqMan 7900HT Sequence Detection System (Applied Biosystems). qPCR gene expression profiling. Intensity of gene expression in CLL cells collected from particular patient before treatment was compared with the gene expression signals in CLL cells from the same patient after 2 weeks of treatment.

ORGANISM(S): Homo sapiens

SUBMITTER: Ida Franiak-Pietryga 

PROVIDER: E-GEOD-33925 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Toward personalized therapy for chronic lymphocytic leukemia: DSC and cDNA microarray assessment of two cases.

Rogalińska Małgorzata M   Franiak-Pietryga Ida I   Błoński Jerzy Z JZ   Góralski Paweł P   Maciejewski Henryk H   Janus Agnieszka A   Robak Paweł P   Mirowski Marek M   Piekarski Henryk H   Robak Tadeusz T   Kiliańska Zofia M ZM  

Cancer biology & therapy 20121031 1

The differences in clinical course of chronic lymphocytic leukemia could have an impact on variations in a patient's response to therapy. Our published results revealed that thermal transition (95 ± 5°C) in differential scanning calorimetry profiles appear to be characteristic for the advanced stage of CLL. Moreover, a decrease/loss of this transition in nuclei from leukemic cells exposed to drugs ex vivo could indicate their diverse efficacy. It seems that the lack of changes in thermal profile  ...[more]

PMID: 23114648

Publication: 1/2

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