Ann Lab Med.  2018 Jan;38(1):63-66. 10.3343/alm.2018.38.1.63.

Molecular and Cytogenetic Studies in a Child with Burkitt Lymphoma and Ataxia-Telangiectasia Syndrome Harboring MYC Overexpression and Partial Trisomy 8

• De Souza M
• Vera-Lozada G
• Othman M
• Marques-Salles TJ
• Pinto LW
• da Rocha M
• Rouxinol S
• Liehr T
• Ribeiro RC
• Hassan R
• Silva ML

• KMID: 2397605
• DOI: http://doi.org/10.3343/alm.2018.38.1.63

Abstract

No abstract available.

MeSH Terms

Ataxia Telangiectasia*
Burkitt Lymphoma*
Child*
Cytogenetics*
Humans
Trisomy*

Figure

• 

  Fig. 1 Cytogenetics and molecular characterization. (A) G-banding cytogenetics: GTG banding karyotype showing the derivative chromosome 8 pointed by the red arrow. (B–F) Molecular cytogenetics: (B) FISH analysis using IGH/MYC/CEP8 Tri-Color Dual Fusion Probe (04N10-020, Abbott Molecular, Des Plaines, IL, USA). Green signal: IGH; red signal: MYC; aqua signal: CEP8; (C) LSI MYC Spectrum Orange Probe (02N22-020, Abbott Molecular) shows derivative chromosome 8 with 2 copies of MYC, 3 in total per cell; (D) Complementary FISH analyses using BCL6 Break Apart Probe (Z-2177-50, ZytoVision GmbH, Bremerhaven, HB, DE) and BLC2 Break Apart Probe (Z-2192-50, ZytoVision GmbH) showed normal partners for both chromosomes 3 and 18, respectively; (E) FISH using partial chromosome paintings for 8p and 8q arms showed partial trisomy 8; (F) Multicolor chromosome banding probe for chromosome 8 characterized the derivative chromosome 8 as a result of t(8;8)(pter->q21::p22->qter); (G–P) Comparisons between cellular genes and microRNA (miRNA) expressions among our patient and classical Burkitt lymphomas, healthy bone marrow (BM) cells, reactive follicular hyperplasias (RFH), and BL- and diffuse large B-cell lymphoma (DLBCL)-derived cell lines. (G) MYC; (H) BCL2; (I) CD10; (J) miR-155; (K) miR-Let7a; (L) miR-Let7b; (M) miR-Let7e; (N) miR-9*; (O) miR-150; (P) miR-21. Case: study patient; BL: classical Burkitt lymphoma; BL-CL: represent the mean values of BL-derived cell lines—Namalwa, Raji, and Ramos; DLBCL-CL: represents the mean values of diffuse large B-cell lymphoma (DLBCL)-derived cell lines—Farage and Pfeiffer; BM: bone marrow cells from healthy donors; RFH: reactive follicular hyperplasia of lymph nodes. The line set at 1 represents the calibration reference. RNA was extracted from FFPE BM biopsy (case) and BL and RFH lymph node using MasterPure™ RNA Purification Kit (Epicentre, Madison, WI, USA). RNA from BM and cell lines was extracted with Direct-zol™ RNA MiniPrep (Zymo Research, Irvine, CA, USA). Relative expression of CD10 and BCL2 was evaluated by TaqMan® assays, as previously described [10], using the average of ACTB and B2M reference genes for normalization. MYC expression was quantified with SYBR green® assays using the average of ACTB and GUSB for normalization. miRNAs were quantified with stem-loop TaqMan® assays (Applied Biosystems, Life Technologies, Carlsbad, CA, USA) after reverse transcription with MicroRNA Reverse Transcription Kit (Applied Biosystems, Life Technologies) for each miRNA and the reference small RNA RNU48. Quantification values were expressed as fold change (2−ΔΔCq) after calibration with the classical BL sample exhibiting the lowest expression level. Bars represent the mean of fold change values in each category, except for the case, in which the mean of two different experiments was represented. Error bars represent standard error of the mean.

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