1. 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.
- Author
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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, and Padua RA
- Subjects
- Aged, Asia, Biopsy, Disease-Free Survival, Europe, Female, Gene Expression Profiling standards, Genetic Predisposition to Disease, Humans, Kaplan-Meier Estimate, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse mortality, Lymphoma, Large B-Cell, Diffuse pathology, Male, Middle Aged, Phenotype, Predictive Value of Tests, Proportional Hazards Models, Reproducibility of Results, South America, Time Factors, Biomarkers, Tumor genetics, Fixatives chemistry, Formaldehyde chemistry, Gene Expression Profiling methods, Lymphoma, Large B-Cell, Diffuse genetics, Paraffin Embedding, Real-Time Polymerase Chain Reaction standards, Reverse Transcriptase Polymerase Chain Reaction standards, Tissue Fixation methods, Transcriptome
- 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.
- Published
- 2016
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