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

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Tekin N., Omidvar N., Morris T. P., Conget P., Bruna F., Timar B., ...Daha Fazla

ONCOTARGET, cilt.7, sa.50, ss.83319-83329, 2016 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 7 Sayı: 50
  • Basım Tarihi: 2016
  • Doi Numarası: 10.18632/oncotarget.13066
  • Dergi Adı: ONCOTARGET
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.83319-83329
  • Anahtar Kelimeler: diffuse large B-cell lymphoma (DLBCL), 6-gene predictor score, formalin fixed paraffin embedded tissue (FFPE), RESIDUAL DISEASE DETECTION, POLYMERASE-CHAIN-REACTION, TIME RT-PCR, CANCER PROGRAM, MODEL, EXPRESSION, SURVIVAL, LEUKEMIA, EUROPE, GENES
  • Ankara Üniversitesi Adresli: Evet

Özet

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.