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Prepublished online as a Blood First Edition Paper on January 2, 2003; DOI 10.1182/blood-2002-08-2577.
NEOPLASIA
From the Abteilung für Pathologie des
Universitätsklinikums Ulm, Germany; Institut
für Humangenetik, Universitätsklinikum Schleswig-Holstein,
Campus Kiel, Germany; Deutsches Krebsforschungszentrum,
Abteilung Molekulare Genetik, Heidelberg, Germany;
Pathologisches Institut der Universität Heidelberg,
Germany; and Institut für Hämatopathologie,
Universitätsklinikum Schleswig-Holstein, Campus Kiel,
Germany.
Structural aberrations of the short arm of chromosome 2, mostly resulting in gains of 2p13~16, have recently been
described as being highly recurrent in Hodgkin and Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL). As these gains consistently lead to increased copy numbers of the REL oncogene locus,
we investigated the expression of the c-Rel protein in a series of 30 cHL cases with known genomic REL status as determined by
comparative genomic hybridization and interphase cytogenetics.
Expression of the c-Rel protein was investigated in 26 biopsies by
immunohistochemistry. Distinct patterns were observed in HRS cells with
no staining, cytoplasmic, and/or nuclear staining for c-Rel. All 13 samples with additional copies of the REL locus displayed
nuclear staining for c-Rel, while 13 cHL samples lacking chromosome 2 (2p) gains displayed a significantly lower proportion or complete
absence of HRS cells with nuclear c-Rel expression. Detailed analysis using combined immunophenotyping and interphase cytogenetics of individual HRS cells demonstrated that REL gains correlated
with the presence of nuclear c-Rel staining. Additionally, in 2 cHL samples with translocation breakpoints in 2p13~16, nuclear staining of c-Rel was observed; in one of them the staining pattern was indicative of a truncated c-Rel protein. The correlation between structural aberrations involving the REL locus and nuclear
c-Rel accumulation in HRS cells qualifies REL as a
target gene of the frequent gains in 2p in cHL. The data suggest that
REL aberrations are a genetic mechanism contributing to
constitutive nuclear factor (NF)- Classical Hodgkin lymphoma (cHL) accounts for
approximately one third of all malignant lymphomas. It is widely
accepted that in most instances the neoplastic Hodgkin and
Reed-Sternberg (HRS) cells represent clonal populations of transformed
germinal center B cells.1,2 The HRS cells are embedded in
a background of reactive cells that are attracted by cytokines produced
by HRS cells.3,4 The reasons for this particular feature
of cHL are largely unknown; however, recent advances in the field have enabled new mechanisms to be identified, such as a constitutive activation of transcription factor nuclear factor (NF)- Very recently, using comparative genomic hybridization (CGH) from
microdissected HRS cells as well as simultaneous fluorescence immunophenotyping and interphase cytogenetics (FICTION;
fluorescence immunophenotyping and interphase
cytogenetics as a tool for
investigation of neoplasms), we
were able to identify structural aberrations affecting the short arm of
chromosome 2 (2p) as the most frequent chromosomal imbalance in cHL.
Changes in 2p, mostly leading to gains, were observed in up to 55% of
all cHL cases, with a predominance in the nodular-sclerosis (cHL-NS)
subtype.7-9
The high frequency of gains of 2p in cHL might indicate that this
chromosomal region harbors one or more genes potentially involved in
the pathogenesis of this disease. The critical region of gain in cHL
was narrowed to 2p13~16 by CGH. The 2 candidate oncogenes
BCL11A and REL have been mapped within this
critical region.9 Using interphase cytogenetics, the HRS
cells of 3 cHL cases with changes in 2p were found to display aberrant
signal patterns only for a fluorescence in situ hybridization (FISH) probe spanning the REL locus but not for a probe spanning
the BCL11A locus.9 These data suggest that
REL, rather than BCL11A, is the target of the 2p
alterations in cHL.
REL encodes a member of the family of NF- We herein show by immunohistochemistry and by simultaneous interphase
cytogenetics and fluorescence immunocytology that structural chromosomal aberrations in 2p correlate with nuclear c-Rel protein accumulation in HRS cells of cHL.
Tissue samples
Immunohistochemistry
For detection of c-Rel protein, the polyclonal rabbit serum 265 raised against a synthetic peptide of the C-terminus of the human c-Rel protein was applied.19,20 Furthermore, in one sample we also applied the serum 1136 raised against amino acid sequence 340-353, which corresponds to a more N-terminally located epitope of human c-Rel.21 Evaluation of immunostaining was carried out in a blinded fashion. In
tissue sections, strongly stained immunoblasts or lymph follicles were
regarded as positive intrinsic controls. Several categories were
created according to the proportion of HRS cells displaying positive
staining for c-Rel: Transfection and blocking experiments To test the specificity of the anti-Rel antisera, transfection experiments using human embryonic kidney cells 293 were carried out. These cells were transiently transfected with the Rc/CMV2 vector and the Rc/CMV2 plasmid with the full-length REL cDNA according to the manufacturer's protocol (Fugene; Roche Biochemicals, Mannheim, Germany). Cells were pelleted and fixed in formalin 48 hours after transfection and embedded in paraffin. Subsequently 2- to 4-µm-thick paraffin sections of the cell pellets were subjected to immunocytological staining (see "Immunohistochemistry").For blocking experiments, specific competing synthetic peptides, originally applied for immunization of rabbits, were used (1 µg peptide/µL serum). Prior to staining, 50 µL of the serum and 50 µL competitive peptide were incubated together on the slides and after incubation for 30 minutes, whereupon the subsequent staining procedure with secondary antibodies was carried out. In parallel, an irrelevant antirabbit serum was used as control. Fluorescence in situ hybridization (FISH) and combined immunophenotyping and interphase cytogenetics (FICTION) Probes for the REL and BCL11A loci used for interphase cytogenetic studies have been described recently.9,22 A chromosome 2 centromeric probe (CEP2) was obtained from Vysis (Downers Grove, IL). The FISH experiments were performed as described elsewhere.23In order to study both c-Rel expression pattern and REL genomic status simultaneously, FICTION was carried out. Touch or cytospin preparations from lymphoma tissues were fixed in acetone for 10 minutes and air dried. Slides were then incubated with the rabbit c-Rel polyclonal antibody 265, followed by sequential application of Alexa 350-conjugated goat antirabbit and Alexa 350-conjugated donkey antigoat secondary antibodies (Molecular Probes, Leiden, the Netherlands). Incubation steps were performed at room temperature with a 30-minute incubation period and subsequent washings. Subsequent to fluorescence immunostaining, slides were incubated for 10 minutes in Carnoy fixative (methanol-acetic acid, 3:1), washed in dH2O and postfixed in 1% paraformaldehyde for 1 minute. Slides were then dehydrated by an ethanol series (70%, 85%, and absolute) and air dried in the dark. The appropriate FISH probe (1.5 µL) was applied to the cell-containing area of the slide and covered with a round 10-mm cover slip. Probe and target DNA were simultaneously denatured at 70°C for 7 minutes and incubated overnight at 37°C. Posthybridization washes were performed in 0.1 × standard saline citrate (SSC) 3 times for 5 minutes each at 60°C. Finally, slides were mounted in antifade solution.24 No DNA counterstaining was performed. Slides were analyzed using a Zeiss Axioskop2 fluorescence microscope (Göttingen, Germany) equipped with the appropriate filter sets (AHF, Tübingen, Germany) and documented using the ISIS imaging system (MetaSystems, Altlussheim, Germany). HRS cells were identified by virtue of their atypical cytology, hyperploidy, and, in positive cases, c-Rel expression. A median of 12 HRS cells (range, 2-31 cells) were evaluated per cHL case. Statistical analysis Correlations between c-Rel expression and genomic REL aberrations in various subgroups were calculated using the Fisher exact test application in SPSS Version 11.0 (SPSS, Erkrath, Germany); P values less than .05 were considered significant. For statistical analyses, only samples taken at diagnosis were considered (ie, case 16b was excluded).
Determination of the genomic REL status Twenty-six biopsies from 25 cHL patients, for which CGH data from microdissected HRS cells were available, were included in the study. Gains of chromosomal material on the short arm of chromosome 2, including the critical region in 2p13~16 containing the REL gene, were detected in 12 cHL biopsies (Table 1). Two of these lymphomas displayed a distinct high-level DNA amplification of the REL locus. In order to assess the REL copy number in these cases, we performed interphase cytogenetics (FISH and FICTION) using probes for the REL locus and the centromeric region of chromosome 2. Seventeen of the 26 biopsies were successfully analyzed in this way. In 9 lymphomas, the balanced status for 2p detected by CGH was also found by interphase cytogenetics (median REL/D2Z1 ratio = 1.0). The 2 cases with high-level amplification by CGH showed high median REL/D2Z1 ratios of 6.0 and 7.5, confirming the presence of an amplification of the REL locus. In 5 cases with gain of 2p by CGH, the median REL/D2Z1 ratio was between 1.3 and 2.0, thus exceeding the balanced status. Overall, there was complete agreement between the findings obtained by the 2 techniques, except in one case: in cHL14, CGH revealed a balanced status while interphase FISH indicated amplification of the REL locus, with a REL/D2Z1 ratio of 5.0.Immunohistochemical detection of the c-Rel protein Specificity of the c-Rel antisera 265 and 1136 applied for immunohistochemistry was proven by transfection experiments in 293 human embryonic kidney cells as well as by blocking experiments with specific peptides in lymphoma tissue (see Figure 1).
Expression of the c-Rel protein in tonsillar tissue and classical Hodgkin lymphoma In hyperplastic tonsillar tissue, a strong, predominantly cytoplasmic staining confined to germinal center cells was obtained with both antisera. Inconsistently, there was a faint nuclear staining, equally confined to germinal center cells. In the extrafollicular areas a small number of intermingled immunoblasts revealed a strong cytoplasmic staining (Figure 2Ai-ii). Because of its superior staining quality, serum 265 was used for all subsequent immunohistochemical and FICTION studies.Staining of the HRS cells revealed 4 different patterns: negative
(Figure 2B), positive in the cytoplasm
(Figure 2C), positive in the nucleus (Figure 2F), and positive in the
cytoplasm and the nucleus (Figure 2Dii,2E). In some lymphomas,
different staining patterns coexisted within the tumor cell population
(Figure 2Hi).
Of the 26 cHL samples with available CGH data, 23 samples from 22 patients could be immunohistochemically evaluated for c-Rel protein expression. All 12 biopsies with gains or amplifications of 2p13~16 by CGH displayed a nuclear staining for c-Rel. The same held true for cHL14, in which amplification of the REL locus was evidenced by interphase FISH. Eleven of these 13 cHL samples showed an additional cytoplasmic staining, while 2 samples with gains on 2p (including 1 sample with a high-level DNA amplification of the REL locus) showed nuclear staining only (nos. 15 and 19). In cHL19, with REL staining restricted to the nucleus, the gain on the short arm of chromosome 2 was the sole imbalance detected by CGH. In order to analyze the correlation between REL copy number and c-Rel protein expression on the single-cell level, FICTION analysis was successfully performed on 7 of the 13 cHL samples with gains in 2p. FICTION confirmed nuclear expression of c-Rel protein in HRS cells with supernumerary copies of the REL locus (Table 1; Figure 2F-G,Hii). FICTION analysis of 3 additional cHL samples with high-level amplification of the REL locus (previous case nos. 1, 2, and 4 in Martin-Subero et al, 2002)9 also confirmed abnormally strong nuclear c-Rel staining (data not shown). Among the 13 cHL samples without gain on 2p according to CGH and interphase cytogenetics, 6 completely lacked c-Rel staining by immunohistochemistry or FICTION. In 2 of these, there was a slight discrepancy between the 2 techniques, most likely due to a very faint c-Rel expression as detected by fluorescence immunophenotyping (cHL1) or a low percentage of c-Rel+ HRS cells (cHL9). Nevertheless, over the complete study there was good overall accordance between immunohistochemistry and FICTION. Eight cHL samples without gains in 2p displayed nuclear staining by immunohistochemistry, including 5 with simultaneous cytoplasmic staining. Remarkably, the percentage of HRS cells with nuclear positivity for c-Rel on immunohistochemistry never exceeded 70% and was below 30% in 7 of 10 cHL samples without gain in 2p. In contrast, the percentage of nuclear c-Rel+ HRS cells in cHL samples with 2p gain was always more than 30% (P = .001) and exceeded 70% in 7 of 12 samples (P = .005; cHL16b excluded). The nuclear c-Rel accumulation (9 of 10 biopsies), like the genomic gain in 2p (8 of 10 biopsies), was particularly prominent in the nodular sclerosis subtype of cHL. In addition to the cases studied by CGH, 2 cHL cases were analyzed in which previous interphase cytogenetics revealed evidence of a breakpoint within the region spanned by the probe for the REL locus.9 Immunohistochemistry of one of these cases, which had been previously shown to contain a t(2;22)(p16;q12),25 featured strong nuclear and cytoplasmic expression of c-Rel as revealed by the antiserum 265 (Figure 2E). The second case with a split of the probe spanning the REL locus completely lacked staining with serum 265. Since a fusion protein lacking the C-terminus of c-Rel has recently been described in the human diffuse large B-cell lymphoma (DLBCL) cell line RC-K8, we also applied the serum 1136, recognizing a more N-terminally located epitope of c-Rel in this latter case.26 Indeed, the HRS cells of this case strongly stained with this serum in the nucleus and cytoplasm, suggesting that the c-Rel protein expressed in these HRS cells lacks the wild-type C-terminus.
In the present study, we show for the first time that HRS cells of most cHL samples express the c-Rel protein. This held particularly true for samples containing chromosomal aberrations in 2p13~16, which in the present series consistently accumulated nuclear c-Rel protein in high percentage of HRS cells. Within the subset of cHL samples with alterations involving the REL locus, the percentage of nuclear c-Rel+ HRS cells was significantly higher than in cHL samples with apparently normal REL loci. These findings suggest that, at least in a subset of cHL, nuclear accumulation of c-Rel expression is related to chromosomal changes affecting the corresponding genomic region. In nonneoplastic lymphoid tissue we found a high c-Rel expression in germinal center cells. These data are in line with mouse models indicating an important role of c-Rel in the development and differentiation of germinal center B cells.27,28 Nevertheless, expression of c-Rel was not restricted to the germinal center but was also observed in a few intermingled immunoblasts in the extrafollicular area. This is in accordance with recent gene-expression profiling data showing strong REL transcript expression in activated B cells.29 Most germinal center cells and immunoblasts expressed c-Rel predominantly in the cytoplasm. Only rarely did we also observe the protein in the nucleus, suggesting a transitory nuclear translocation of c-Rel protein during germinal center B-cell reaction. The most striking finding of the present study was the nuclear
accumulation of c-Rel in a high percentage of HRS cells in cHL samples
with genomic changes involving the REL oncogene locus. In
the clear majority of cHL cases hitherto characterized, chromosomal alterations in the short arm of chromosome 2 led to gain of the REL gene. In these lymphomas, nuclear c-Rel accumulation
might be caused by a gene dosage effect analogous to the mechanisms described for numerous other oncogenes, such as HER2/NEU,
BCL2, and CCNE.30-32 In 2 cHL
samples with nuclear c-Rel accumulation, we detected chromosomal
breakpoints, using a FISH probe spanning the REL locus. One
of those cases carried a translocation t(2;22), cytogenetically
suggesting deregulation of c-Rel through juxtaposition of the
REL gene and regulatory sequences of the immunoglobulin light chain locus In the second cHL sample with a break event detected by the REL-spanning FISH probe, the precise chromosomal rearrangement affecting the REL region is not yet clear.9 Strikingly, this case displayed strong nuclear staining using an antiserum directed to the REL homology domain recognizing amino acids 340 through 353. In contrast, no staining was observed using the serum 265 targeting amino acids 573 through 587 of the human c-Rel protein. These findings can be explained by the presence of a truncation of the C-terminus of the wild-type c-Rel protein caused by the chromosomal change. Such a mechanism of c-Rel activation has recently been demonstrated in the cell line RC-K8, derived from a human large B-cell lymphoma. In this cell line, the 5' end of the REL gene is fused with the 3' end of another gene called NRG (non-REL gene), resulting in a chimeric protein devoid of the wild-type C-terminus of c-Rel.26 Apart from the cell line RC-K8, genomic changes affecting the REL locus have been described in numerous primary lymphomas. In particular, gains including the REL locus have previously been reported in other B-cell lymphomas, such as diffuse large B-cell lymphoma,35,36 mediastinal B-cell lymphoma,37 and follicular lymphomas.38 Nevertheless, the frequency of 2p gains in these non-Hodgkin lymphomas did not exceed 30% in most series and is thus considerably lower than in cHL. Moreover, genomic amplification of REL was recently shown to be restricted to the germinal center B cell-like subtype of DLBCLs defined by lymphochip technology.39 Finally, a recent abstract reports that in DLBCL, nuclear expression of c-Rel correlated with REL gene amplification.40 A well-known common characteristic of HRS cells is a high level of
constitutive nuclear NF- We conclude that, based on the close correlation between
structural chromosomal aberrations in 2p13~16 and nuclear
accumulation of c-Rel described in the present study, it seems likely
that genomic changes of the REL locus provide an alternative
genetic mechanism of constitutive nuclear NF-
We wish to thank Dr N. Rice and Dr M. Ernst, National Cancer Institute - Frederick Cancer Research and Development Center (NCI-FCRDC), Frederick, MD, for helpful discussion and for providing the c-Rel antisera, the competing peptides, and the c-Rel vector. We thank Iwona Nerbas, Claudia Becher, and Dorit Schuster for excellent technical assistance and Caroline Higginson for editorial help.
Submitted August 22, 2002; accepted December 23, 2002.
Prepublished online as Blood First Edition Paper, January 2, 2003; DOI 10.1182/blood-2002-08-2577.
Supported by the Tumorzentrum Heidelberg/Mannheim (I.I.2.), the Deutsche Krebshilfe (70-2859-Ba2), the Landesforschungschwerpunkt Baden Württemberg (D.1526.1.4), and the Interdisziplinäre Zentrum für klinische Krebsforschung (IZKF) Kiel.
T.F.E.B. and J.I.M.-S. contributed equally to this work.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Peter Möller, Abteilung für Pathologie, Universität Ulm, Albert-Einstein-Allee 11, D-89081 Ulm, Germany; e-mail: peter.moeller{at}medizin.uni-ulm.de; or Reiner Siebert, Institut für Humangenetik, Universitätsklinikum Schleswig-Holstein, Campus Kiel, Schwanenweg 24, D-24105 Kiel, Germany; e-mail: rsiebert{at}medgen.uni-kiel.de.
1. Küppers R, Schwering I, Bräuninger A, Rajewsky K, Hansmann ML. Biology of Hodgkin's lymphoma. Ann Oncol. 2002;13(suppl 1):11-18[Abstract].
2.
Marafioti T, Hummel M, Foss HD, et al.
Hodgkin and Reed-Sternberg cells represent an expansion of a single clone originating from a germinal center B-cell with functional immunoglobulin gene rearrangements but defective immunoglobulin transcription.
Blood.
2000;95:1443-1450
3.
Staudt LM.
The molecular and cellular origins of Hodgkin's disease.
J Exp Med.
2000;191:207-212
4.
Skinnider BF, Mak TW.
The role of cytokines in classical Hodgkin lymphoma.
Blood.
2002;99:4283-4297 5. Bargou RC, Emmerich F, Krappmann D, et al. Constitutive nuclear factor-kappaB-RelA activation is required for proliferation and survival of Hodgkin's disease tumor cells. J Clin Invest. 1997;100:2961-2969[Medline] [Order article via Infotrieve].
6.
Hinz M, Loser P, Mathas S, Krappmann D, Dörken B, Scheidereit C.
Constitutive NF-
7.
Joos S, Küpper M, Ohl S, et al.
Genomic imbalances including amplification of the tyrosine kinase gene JAK2 in CD30+ Hodgkin cells.
Cancer Res.
2000;60:549-552
8.
Joos S, Menz CK, Wrobel G, et al.
Classical Hodgkin lymphoma is characterized by recurrent copy number gains of the short arm of chromosome 2.
Blood.
2002;99:1381-1387
9.
Martin-Subero JI, Gesk S, Harder L, et al.
Recurrent involvement of the REL and BCL11A loci in classical Hodgkin lymphoma.
Blood.
2002;99:1474-1477 10. Baldwin AS Jr. The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol. 1996;14:649-683[CrossRef][Medline] [Order article via Infotrieve]. 11. Gilmore TD. The Rel/NF-kappaB signal transduction pathway: introduction. Oncogene. 1999;18:6842-6844[CrossRef][Medline] [Order article via Infotrieve]. 12. Rayet B, Gelinas C. Aberrant rel/nfkb genes and activity in human cancer. Oncogene. 1999;18:6938-6947[CrossRef][Medline] [Order article via Infotrieve]. 13. Siebenlist U, Franzoso G, Brown K. Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol. 1994;10:405-455[CrossRef][Medline] [Order article via Infotrieve]. 14. Wulczyn FG, Krappmann D, Scheidereit C. The NF-kappa B/Rel and I kappa B gene families: mediators of immune response and inflammation. J Mol Med. 1996;74:749-769[CrossRef][Medline] [Order article via Infotrieve]. 15. Cabannes E, Khan G, Aillet F, Jarrett RF, Hay RT. Mutations in the IkBa gene in Hodgkin's disease suggest a tumour suppressor role for IkappaBalpha. Oncogene. 1999;18:3063-3070[CrossRef][Medline] [Order article via Infotrieve].
16.
Jungnickel B, Staratschek-Jox A, Bräuninger A, et al.
Clonal deleterious mutations in the IkappaBalpha gene in the malignant cells in Hodgkin's lymphoma.
J Exp Med.
2000;191:395-402 17. Wood KM, Roff M, Hay RT. Defective IkappaBalpha in Hodgkin cell lines with constitutively active NF-kappaB. Oncogene. 1998;16:2131-2139[CrossRef][Medline] [Order article via Infotrieve]. 18. Jaffe ES, Harris NL, Stein H, Vardiman JW. Tumours of Haematopoietic and Lymphoid Tissues. Lyon, France: IARC Press; 2001. 19. Whiteside ST, Epinat JC, Rice NR, Israel A. I kappa B epsilon, a novel member of the I kappa B family, controls RelA and cRel NF-kappa B activity. EMBO J. 1997;16:1413-1426[CrossRef][Medline] [Order article via Infotrieve]. 20. Rice NR, MacKichan ML, Israel A. The precursor of NF-kappa B p50 has I kappa B-like functions. Cell. 1992;71:243-253[CrossRef][Medline] [Order article via Infotrieve].
21.
Latimer M, Ernst MK, Dunn LL, Drutskaya M, Rice NR.
The N-terminal domain of IkappaB alpha masks the nuclear localization signal(s) of p50 and c-Rel homodimers.
Mol Cell Biol.
1998;18:2640-2649
22.
Satterwhite E, Sonoki T, Willis TG, et al.
The BCL11 gene family: involvement of BCL11A in lymphoid malignancies.
Blood.
2001;98:3413-3420 23. Schlegelberger B, Metzge S, Harder S, Zühlke-Jenisch R, Zhang Y, Siebert R. Classical and molecular cytogenetics of tumor cells. In: Wegner RD, ed. Diagnostic Cytogenetics. Heidelberg, Germany: Springer-Verlag; 1999:151-185.
24.
Martin-Subero JI, Chudoba I, Harder L, et al.
Multicolor-FICTION: expanding the possibilities of combined morphologic, immunophenotypic, and genetic single cell analyses.
Am J Pathol.
2002;161:413-420 25. Schlegelberger B, Weber-Matthiesen K, Himmler A, et al. Cytogenetic findings and results of combined immunophenotyping and karyotyping in Hodgkin's disease. Leukemia. 1994;8:72-80[Medline] [Order article via Infotrieve]. 26. Kalaitzidis D, Gilmore TD. Genomic organization and expression of the rearranged REL proto-oncogene in the human B-cell lymphoma cell line RC-K8. Genes Chromosomes Cancer. 2002;34:129-135[CrossRef][Medline] [Order article via Infotrieve]. 27. Gugasyan R, Grumont R, Grossmann M, et al. Rel/NF-kappaB transcription factors: key mediators of B-cell activation. Immunol Rev. 2000;176:134-140[CrossRef][Medline] [Order article via Infotrieve]. 28. Tumang JR, Owyang A, Andjelic S, et al. c-Rel is essential for B lymphocyte survival and cell cycle progression. Eur J Immunol. 1998;28:4299-4312[CrossRef][Medline] [Order article via Infotrieve]. 29. Alizadeh AA, Eisen MB, Davis RE, et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature. 2000;403:503-511[CrossRef][Medline] [Order article via Infotrieve].
30.
Monni O, Joensuu H, Franssila K, Klefstrom J, Alitalo K, Knuutila S.
BCL2 overexpression associated with chromosomal amplification in diffuse large B-cell lymphoma.
Blood.
1997;90:1168-1174 31. Schwab M. Oncogene amplification in solid tumors. Semin Cancer Biol. 1999;9:319-325[CrossRef][Medline] [Order article via Infotrieve].
32.
Stark A, Hulka BS, Joens S, et al.
HER-2/neu amplification in benign breast disease and the risk of subsequent breast cancer.
J Clin Oncol.
2000;18:267-274 33. Siebert R, Rosenwald A, Staudt LM, Morris SW. Molecular features of B-cell lymphoma. Curr Opin Oncol. 2001;13:316-324[CrossRef][Medline] [Order article via Infotrieve].
34.
Willis TG, Dyer MJ.
The role of immunoglobulin translocations in the pathogenesis of B-cell malignancies.
Blood.
2000;96:808-822 35. Barth TF, Bentz M, Leithäuser F, et al. Molecular-cytogenetic comparison of mucosa-associated marginal zone B-cell lymphoma and large B-cell lymphoma arising in the gastro-intestinal tract. Genes Chromosomes Cancer. 2001;31:316-325[CrossRef][Medline] [Order article via Infotrieve].
36.
Houldsworth J, Mathew S, Rao PH, et al.
REL proto-oncogene is frequently amplified in extra-nodal diffuse large cell lymphoma.
Blood.
1996;87:25-29 37. Bentz M, Barth TF, Brüderlein S, et al. Gain of chromosome arm 9p is characteristic of primary mediastinal B-cell lymphoma (MBL): comprehensive molecular cytogenetic analysis and presentation of a novel MBL cell line. Genes Chromosomes Cancer. 2001;30:393-401[CrossRef][Medline] [Order article via Infotrieve]. 38. Goff LK, Neat MJ, Crawley CR, et al. The use of real-time quantitative polymerase chain reaction and comparative genomic hybridization to identify amplification of the REL gene in follicular lymphoma. Br J Haematol. 2000;111:618-625[CrossRef][Medline] [Order article via Infotrieve].
39.
Rosenwald A, Wright G, Chan WC, et al.
The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma.
N Engl J Med.
2002;346:1937-1947 40. Donelly GB, Teruya-Feldstein J, Quin J, Zelenetz AD, Houldsworth J, Chaganti RS. Nuclear expression of Rel correlates with gene amplification in diffuse large B-cell lymphomas (DLBCL) [abstract]. Blood. 2001;98:302. 41. Mathas S, Hinz M, Anagnostopoulos I, et al. Aberrantly expressed c-Jun and JunB are a hallmark of Hodgkin lymphoma cells, stimulate proliferation and synergize with NF-kappaB. EMBO J. 2002;21:4104-4113[CrossRef][Medline] [Order article via Infotrieve].
42.
Skinnider BF, Elia AJ, Gascoyne RD, et al.
Signal transducer and activator of transcription 6 is frequently activated in Hodgkin and Reed-Sternberg cells of Hodgkin lymphoma.
Blood.
2002;99:618-626
© 2003 by The American Society of Hematology.
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  N. Gupta, J. Delrow, A. Drawid, A. M. Sengupta, G. Fan, and C. Gelinas Repression of B-Cell Linker (BLNK) and B-Cell Adaptor for Phosphoinositide 3-Kinase (BCAP) Is Important for Lymphocyte Transformation by Rel Proteins Cancer Res., February 1, 2008; 68(3): 808 - 814. [Abstract] [Full Text] [PDF]
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 G Kapatai and P Murray Contribution of the Epstein Barr virus to the molecular pathogenesis of Hodgkin lymphoma J. Clin. Pathol., December 1, 2007; 60(12): 1342 - 1349. [Abstract] [Full Text] [PDF]
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Abstract
Introduction
B/Rel activation in cHL.
(Blood. 2003;101:3681-3686)
have been described in HRS cells of a small subset
of cHL and might account for NF-
stands for no staining detected, + indicates
staining in up to 30%, ++ indicates staining in more than 30% and up
to 70%, and +++ indicates staining in more than 70% of the total
number of HRS cells analyzed (Table 
(Ai). In the extrafollicular region (Aii),
intermingled immunoblasts are c-Rel+; bars = 37.5µm.
(B) In cHL1, with balanced genomic status for REL by CGH and
FICTION, the HRS cells are c-Rel protein
(IGL) in 22q.