Selection of reference genes for quantitative PCR studies in purified B cells from B cell chronic lymphocytic leukaemia patients

Vilma Valceckiene (1) , Rima Kontenyte (1) , Arturas Jakubauskas (1) , Laimonas Griskevicius (2)

1 Hematology, Oncology and Transfusion Medicine Center,Department of Molecular and Regenerative Medicine
2 Hematology, Oncology and Transfusion Medicine Center, Department of Hematology

Clinical heterogeneity of B-cell chronic lymphocytic leukaemia (B-CLL) makes it necessary to identify potent prognostic indicators to predict individual clinical course and select risk-adapted therapy. During the last years numerous gene expression models have been suggested as prognostic factors of B-CLL. Today quantitative polymerase chain reaction (qPCR) is a preferred method for rapid quantification of gene expression and validation of microarray data. Reliability of qPCR data is highly dependent on the use of appropriate reference genes for normalisation. To date, no validated reference genes have been reported for normalisation of gene expression in B-CLL. Therefore, the present study was conducted to identify suitable reference genes for gene expression studies in CD19+ B cells isolated from B-CLL patients' peripheral blood. The stability of ACTB, B2M, GAPDH, GUSB, HMBS, HPRT1, MRPL19, TBC and UBC genes was determined by three different descriptive statistics, geNorm, NormFinder and BestKeeper-1, which produced highly comparable results. Based on our results, B2M, HPRT1, and GUSB were found to be the most suitable reference genes for qPCR studies in B-CLL patients' peripheral blood B cells.

Mots clés: Medicine

Vilma Valceckiene, Rima Kontenyte, Arturas Jakubauskas, Laimonas Griskevicius. Selection of reference genes for quantitative PCR studies in purified B cells from B cell chronic lymphocytic leukaemia patients. British Journal of Haematology, 2010, 151 (3), pp.232. ⟨10.1111/j.1365-2141.2010.08363.x⟩. ⟨hal-00573092⟩