unfavorable prognostic factor

MYC and BCL2 Protein Expression in Patients with Diffuse Large B-cell Lymphoma

LYMPHOID TUMORS , , , ,

AE Misyurina1, AM Kovrigina1, EA Baryakh1, VA Misyurin2, SK Kravchenko1, AV Misyurin2, TN Obukhova1, SM Kulikov1, AN Kopylov2, AU Magomedova1, EG Gemdzhyan1, AI Vorob’ev1

1 Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167

2 N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478

For correspondence: Anna Evgen’evna Misyurina, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167; Tel.: +7(909)637-32-46; e-mail: anna.lukina1@gmail.com

For citation: Misyurina AE, Kovrigina AM, Baryakh EA, et al. MYC and BCL2 Protein Expression in Patients with Diffuse Large B-cell Lymphoma. Clinical oncohematology. 2015;8(1):44–53 (In Russ).

ABSTRACT

Objective. The objective of this study is to analyze the incidence and the role of MYC and BCL2 protein expression in patients with diffuse large B-cell lymphoma (DLBCL) and to compare data of histological, immunohistochemical, genetic, and moleculobiological analyses with clinical characteristics.

Methods. 62 patients with confirmed diffuse large B-cell lymphoma (DLBCL) were enrolled in the study; they underwent treatment according to the original modified protocol NHL-BFM-90 ± R in the Hematology Research Center under the Ministry of Health of the Russian Federation. A reference group consisted of 13 DLBCL patients who underwent СНОР-like ± R therapy. In all observations, histological and immunohistochemical tests were performed in archived biopsy samples of a tumor tissue or a lymph node (paraffin blocks) using BCL2 (clone 124, Dako) and MYC antibodies (clone Y69, Epitomics). Based on C.P. Hans’s algorithm (2004), GCB and non-GCB immunohistochemical subtypes of DLBCL were determined. Standard cytogenetic tests (n = 22) and FISH analysis (n = 52) were performed in this study in order to determine locus translocation of с-MYC gene, IgH gene, t(8;14)(q24;q32), BCL2 gene, t(14;18)(q32;q21). Quantitative RT-PCR on paraffin tumor/node biopsy samples was also performed to evaluate the number of mRNAs of с-MYC and BCL2 genes.

Results. MYC expression was found in 24 (39 %) of 62 DLBCL patients and BCL2 in 36 (58 %) of 62 patients (threshold values were 40 % and 50 % of tumor cells, respectively). MYC/BCL2 coexpression was diagnosed in 15 (24 %) of 62 DLBCL patients. 4 (27 %) of 15 patients with MYC/BCL2 coexpression had a GCB-subtype of DLBCL, 73 % patients with MYC/BCL2 coexpression were diagnosed with non-GCB subtype of DLBCL (p < 0.02). c-MYC rearrangement was found in two cases (3 %), one of them had a MYC protein expression score more than 40 %. 10 (19 %) patients had one or more additional signal from 8q24 locus of c-MYC gene. We didn’t find any correlation between the presence of additional signals from c-MYC gene locus and the level of immunohistochemical expression of MYC protein ³ 40 % (p < 0.05). BCL2 gene rearrangement was detected in one case; it was accompanied by BCL2 protein immunohistochemical expression ³ 50 %. BCL2 amplification was observed in 17 (40 %) patients. There was a correlation between the amplification of BCL2 gene and immunohistochemical expression of BCL2 protein (threshold value was ³ 50 % of positive cells) (p = 0.0053). There was a direct correlation between the amount of mRNAs and MYC protein (correlation coefficient 0.86; p < 0.0001). There was no correlation between the level of BCL2 gene expression and the amount of protein (correlation coefficient 0.14; p = 0.57). Four-year overall survival for DLBCL patients treated with m-NHL-BFM-90 ± R was 71 % (in patients without MYC/BCL2 coexpression) vs 57 % (in patients with MYC/BCL2 coexpression) (p = 0.39). The probability of relapse or progression of DLBCL was significantly higher in patients with MYC/BCL2 coexpression, than in patients without MYC/BCL2 coexpression (65 % vs 15 %; p = 0.0029).

Conclusions. The MYC/BCL2 coexpression is observed mainly in DLBCL patients with non-GCB immunohistochemical subtype of the disease. The MYC/BCL2 coexpression is vital for prediction of the risk of relapses/progression of the disease in patients receiving an intensive chemotherapy according to the m-NHL-BFM-90 ± R protocol. Taking into account a relatively stable structure of the protein substrate, the obtained data may become a basis for development of a diagnostic immunohistochemical algorithm for stratification of DLBCL patients.

Keywords: DLBCL, intensive therapy, MYC/BCL2 coexpression, immunohistochemistry, unfavorable prognostic factor.

Received: November 8, 2014

Accepted: November 11, 2014

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