Protocol for qRT-PCR analysis from formalin fixed paraffin embedded tissue sections from diffuse large b-cell lymphoma: Validation of the six-gene predictor score

Date

2016

Type:

Artículo

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11

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Journal Title

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Volume Title

Publisher

Impact Journals

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Abstract

As a part of an international study on the molecular analysis of Diffuse Large B-cell Lymphoma (DLBCL), a robust protocol for gene expression analysis from RNA extraction to qRT-PCR using Formalin Fixed Paraffin Embedded tissues was developed. Here a study was conducted to define a strategy to validate the previously reported 6-gene (LMO2, BCL6, FN1, CCND2, SCYA3 and BCL2) model as predictor of prognosis in DLBCL. To avoid variation, all samples were tested in a single centre and single platform. This study comprised 8 countries (Brazil, Chile, Hungary, India, Philippines, S. Korea, Thailand and Turkey). Using the Kaplan-Meier and log rank test on patients (n=162) and two mortality risk groups (with those above and below the mean representing high and low risk groups) confirmed that the 6-gene predictor score correlates significantly with overall survival (OS, p<0.01) but not with event free survival (EFS, p=0.18). Adding the International Prognostic Index (IPI) shows that the 6-gene predictor score correlates significantly with high IPI scores for OS (p<0.05), whereas those with low IPI scores show a trend not reaching significance (p=0.08). This study defined an effective and economical qRT-PCR strategy and validated the 6-gene score as a predictor of OS in an international setting.

Description

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Citation

Tekin N, Omidvar N, Morris TP, Conget P, Bruna F, Timar B, Gagyi E, Basak R, Naik O, Auewarakul C, Sritana N, Levy D, Cerci JJ, Bydlowski SP, Pereira J, Dimamay MP, Natividad F, Chung JK, Belder N, Kuzu I, Paez D, Dondi M, Carr R, Ozdag H, Padua RA. Protocol for qRT-PCR analysis from formalin fixed paraffin embedded tissue sections from diffuse large b-cell lymphoma: Validation of the six-gene predictor score. Oncotarget. 2016 Dec 13;7(50):83319-83329

Keywords

diffuse large B-cell lymphoma (DLBCL), 6-gene predictor score, formalin fixed paraffin embedded tissue (FFPE)

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