Bcl-6

(Bcl-6 protein)
Characteristics
The Bcl-6 protein (B-cell CLL/lymphoma 6, zinc finger protein 51) is a nuclear zinc finger transcription factor with an N-terminal POZ domain. Bcl-6 functions as a transcriptional repressor and is necessary for germinal centre formation. The gene is located at chromosome 3q27. Two alternatively spliced transcript variants that encode the identical protein have been reported for this gene. The Bcl-6 protein acts as a sequence-specific repressor of transcription, and has been shown to modulate the transcription of START-dependent IL-4 responses of B-cells. It can interact with a variety of POZ-containing proteins that function as transcription co-repressors. While it was generally known that this protein is a repressor of transcription, only recently it was highlighted that Bcl-6 suppresses the expression of the p53 gene and modulates DNA damage-induced apoptotic responses in germinal centre B-cells. Bcl-6 represses p53 transcription by binding 2 specific DNA sites within the p53 promoter region and accordingly, p53 expression is absent in germinal centre B-cells where Bcl-6 is highly expressed. Most notably, constitutive expression of Bcl-6 protects B-cell lines from apoptosis induced by DNA damage. These results suggest that an important function of Bcl-6 is to allow germinal centre B-cells to tolerate the physiologic DNA breaks required for immunoglobulin class switch recombination and somatic hypermutation without inducing a p53-dependent apoptotic response. These findings also imply that deregulated Bcl-6 expression contributes to lymphomagenesis in part by functional inactivation of p53. Bcl-6 appears to inhibit differentiation of germinal centre B-cells toward plasma cells by binding to signal transducers and activators of transcription 3, thereby preventing the expression of Blimp-1 (a major regulator of plasma cell development). Mutations of the Bcl-6 gene have been found in normal germinal centre B-cells, and even though it is not clear what role they play in lymphomagenesis, it appears that they may disrupt its negative autoregulation and in this manner promote lymphoma growth. In non-neoplastic lymphoid tissues, Bcl-6 protein is expressed in germinal centre B-cells in secondary follicles in both the dark (centroblast-rich) and light (centrocyte-rich) zones. It is also detectable in a few CD4-positive T cells, both in the germinal centres and in interfollicular areas, and in perifollicular CD30+ lymphoid cells. In addition, Bcl-6 protein is expressed in thymocytes, mainly in the cortex. It was recently also demonstrated in monocytes, but in levels lower than that usually observed in germinal centre B-cells. Bcl-6 is also observed in the squamous epithelium of the epidermis, transitional epithelium and olfactory sensory neurons. In all positive cells, Bcl-6 protein localizes to the nucleus in a diffuse or microgranular pattern.
Neoplasms
In malignant lymphomas, Bcl-6 protein is most commonly detected in germinal centre neoplasms including follicular lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma (40%), lymphocyte predominant Hodgkin lymphoma, and even in peripheral T-cell lymphoma with follicular localization (“follicular T cell lymphoma”). It is also found in CD30+ anaplastic large cell lymphoma. Bcl-6 expression is virtually absent in acute lymphatic leukaemia and mantle cell lymphoma. Translocations of the regulatory part of the Bcl-6 gene juxtapose the gene to promoters from a variety of partner chromosomes (most commonly in Ig chain loci).
Application
Bcl-6 is mainly used in the classification of B-cell lymphomas (see above).
Controls
Normal tonsil is recommended as positive and negative tissue control. Virtually all the germinal centre Bcells must show a moderate to strong nuclear staining reaction, while an at least weak to moderate nuclear staining reaction must be seen in the majority of squamous epithelial cells. In the mantle zones and interfollicular areas only dispersed cells must show a nuclear staining reaction.
Selected references
1. www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=gene&cmd=Retrieve&dopt=full_report&list_uids=604 2. www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=109565 3. Phan RT, Dalla-Favera R. The BCL6 proto-oncogene suppresses p53 expression in germinal-centre B cells. Nature 2004;432: 635-639. 4. Reljic R, Wagner SD, Peakman LJ, Fearon DT. Suppression of signal transducer and activator of transcription 3-dependent B lymphocyte terminal differentiation by BCL-6. J Exp Med. 2000;192:1841-1848. 5. Ye BH, Lista F, Lo Coco F, et al. Alterations of a zinc finger-encoding gene, BCL-6, in diffuse large-cell lymphoma. Science. 1993;262:747-750. 6. Lo Coco F, Ye BH, Lista F, et al. Rearrangements of the BCL6 gene in diffuse large cell non-Hodgkin's lymphoma. Blood. 1994;83:1757-1759. 7. Capello D, Vitolo U, Pasqualucci L, et al. Distribution and pattern of BCL-6 mutations throughout the spectrum of B-cell neoplasia. Blood. 2000;95:651-659. 8. Pasqualucci L, Migliazza A, Fracchiolla N, et al. BCL-6 mutations in normal germinal center B cells: evidence of somatic hypermutation acting outside Ig loci. Proc Natl Acad Sci U S A. 1998;95:11816-11821. 9. Pasqualucci L, Migliazza A, Basso K, Houldsworth J, Chaganti RS, Dalla-Favera R. Mutations of the BCL6 proto-oncogene disrupt its negative autoregulation in diffuse large B-cell lymphoma. Blood. 2003 Apr 15;101(8):2914-23. Epub 2002 Dec 19. 10. Flenghi L, Bigerna B, Fizzotti M, et al. Monoclonal antibodies PG-B6a and PG-B6p recognize, respectively, a highly conserved and a formol-resistant epitope on the human BCL-6 protein amino-terminal region. Am J Pathol. 1996;148:1543-1555. 11. Cattoretti G, Chang CC, Cechova K, et al. BCL-6 protein is expressed in germinal-center B cells. Blood. 1995;86:45-53. 12. Flenghi L, Ye BH, Fizzotti M, et al. A specific monoclonal antibody (PG-B6) detects expression of the BCL-6 protein in germinal center B cells. Am J Pathol. 1995;147:405-411. 13. Onizuka T, Moriyama M, Yamochi T, et al. BCL-6 gene product, a 92- to 98-kD nuclear phosphoprotein, is highly expressed in germinal center B cells and their neoplastic counterparts. Blood. 1995;86:28-37. 14. Falini B, Fizzotti M, Pileri S, Liso A, Pasqualucci L, Flenghi L. BCL-6 protein expression in normal and neoplastic lymphoid tissues. Ann Oncol. 1997;8:101-104. 15. Hyjek E, Chadburn A, Liu YF, Cesarman E, Knowles DM. BCL-6 protein is expressed in precursor T-cell lymphoblastic lymphoma and in prenatal and postnatal thymus. Blood. 2001;97:270-276. 16. Toney LM, Cattoretti G, Graf JA, Merghoub T, Pandolfi PP, Dalla-Favera R, Ye BH, Dent AL. BCL-6 regulates chemokine gene transcription in macrophages. Nat Immunol. 2000 Sep;1(3):214-20. 17. Z, Kim H, Park H, Kim Y, Cheon J, Kim I. The expression of bcl-2 and bcl-6 protein in normal and malignant transitional epithelium.Urol Res. 2003 Aug;31(4):272-5. Epub 2003 Jul 16. 18. Kanazawa N, Moriyama M, Onizuka T, Sugawara K, Mori S. Expression of bcl-6 protein in normal skin and epidermal neoplasms. Pathol Int. 1997 Sep;47(9):600-7. 19. Otaki JM, Fearon DT, Yamamoto H. The proto-oncogene BCL-6 is expressed in olfactory sensory neurons. Neurosci Res. 2005 Oct;53(2):189-200. 20. Bos R, van Diest PJ, van der Groep P, Greijer AE, Hermsen MA, Heijnen I, Meijer GA, Baak JP, Pinedo HM, van der Wall E, Shvarts A. Protein expression of B-cell lymphoma gene 6 (BCL-6) in invasive breast cancer is associated with cyclin D1 and hypoxia-inducible factor-1alpha (HIF-1alpha). Oncogene. 2003 Dec 4;22(55):8948-51. 21. Skinnider BF, Horsman DE, Dupuis B, Gascoyne RD. Bcl-6 and Bcl-2 protein expression in diffuse large B-cell lymphoma and follicular lymphoma: correlation with 3q27 and 18q21 chromosomal abnormalities. Hum Pathol. 1999;30:803-808. 22. Wlodarska I, Nooyen P, Maes B, Martin-Subero JI, Siebert R, Pauwels P, De Wolf-Peeters C, Hagemeijer A. Frequent occurrence of BCL6 rearrangements in nodular lymphocyte predominance Hodgkin lymphoma but not in classical Hodgkin lymphoma. Blood. 2003 Jan 15;101(2):706-10. Epub 2002 Sep 12. 23. Carbone A, Gloghini A, Gaidano G, Dalla-Favera R, Falini B. BCL-6 protein expression in human peripheral T-cell neoplasms is restricted to CD30+ anaplastic large-cell lymphomas. Blood. 1997 Sep 15;90(6):2445-50. 24. de Leval L, Savilo E, Longtine J, Ferry JA, Harris NL. Peripheral T-cell lymphoma with follicular involvement and a CD4+/bcl-6+ phenotype. Am J Surg Pathol. 2001 Mar;25(3):395-400. 25. Raible MD, Hsi ED, Alkan S. Bcl-6 protein expression by follicle center lymphomas. A marker for differentiating follicle center lymphomas from other low-grade lymphoproliferative disorders. Am J Clin Pathol. 1999 Jul;112(1):101-7. 26. Carbone A, Gloghini A, Gaidano G, Dalla-Favera R, Falini B. BCL-6 protein expression in human peripheral T-cell neoplasms is restricted to CD30+ anaplastic large-cell lymphomas. Blood. 1997;90:2445-2450. 27. Garcia JF, Garcia JF, Maestre L, Lucas E, Sanchez-Verde L, Romero-Chala S, Piris MA, Roncador G. Genetic immunization: a new monoclonal antibody for the detection of BCL-6 protein in paraffin sections. J Histochem Cytochem. 2006 Jan;54(1):31-8. Epub 2005 Jul 26.
09.12.14 - ET/MV/LE