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Use of archival formalin-fixed, paraffin-embedded (FFPE) tissue samples for molecular genetic analysis in diffuse large B-cell lymphoma (DLBCL)


Authors: Marie Jarošová;  Jana Kučerová;  Patrik Flodr;  Michaela Mikešová;  Vít Procházka;  Tomáš Papajík
Authors‘ workplace: Hemato-onkologická klinika FN a LF UP Olomouc 1;  Ústav biologie LF UP Olomouc 2;  Ústav klinické a molekulární patologie FN a LF UP Olomouc 3
Published in: Čes.-slov. Patol., 50, 2014, No. 2, p. 95-99
Category: Original Article

Overview

The currently valid molecular genetic subclassification of patients with diffuse large B-cell lymphoma (DLBCL) into three prognostic subgroups based on expression profiling has been the objective of numerous genetic studies. In routine clinical practice, however, expression profiling technology remains unavailable for the most of centers. Apart from the technology, in some cases molecular genetic laboratories have problems obtaining high-quality material, i.e. fresh tissues, for RNA isolation to determine gene expression. One possibility is to determine the gene expression from RNA obtained by isolation from formalin-fixed, paraffin-embedded (FFPE) tissue. This pilot study aimed at isolating RNA from FFPE in patients diagnosed with DLBCL and verifying the potential use of such RNA for the expression analysis of 7 selected genes. Although the study showed that it is possible to isolate RNA and determine the expression of the selected genes from archival material, the values of relative expression of some genes in the set were too variable to be used for unambiguous prognostic classification. It was confirmed that retrospective analyses of selected genes may be performed with sufficient material obtained, and that properly archived blocks may be used for molecular biology analyses even after 8 years.

Keywords:
DLBCL – FFPE – gene expression – qRT-PCR – genetic changes


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