This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Akasaka, T. Articles by Ohno, H. Search for Related Content PubMed PubMed Citation Articles by Akasaka, T. Articles by Ohno, H. Social Bookmarking                            What's this?

Molecular and Clinical Features of Non-Burkitt’s, Diffuse Large-Cell Lymphoma of B-Cell Type Associated With the c-MYC/Immunoglobulin Heavy-Chain Fusion Gene

From the First Division, Department of Internal Medicine, and Laboratory of Anatomical Pathology, Faculty of Medicine, The Center for Molecular Biology and Genetics, Kyoto University, Kyoto; and First Department of Internal Medicine, Kansai Medical University, Moriguchi, Japan.

Address reprint requests to Hitoshi Ohno, MD, First Division, Department of Internal Medicine, Faculty of Medicine, Kyoto University, 54 Shogoin-Kawaramachi, Sakyo-ku, Kyoto 606-8507, Japan; email hohno{at}kuhp.kyoto-u.ac.jp

	ABSTRACT

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

 
PURPOSE: t(8;14)(q24;q32) and/or c-MYC/immunoglobulin heavy-chain (IGH) fusion gene have been observed not only in Burkitt’s lymphoma (BL) but also in a proportion of non-BL, diffuse large-cell lymphoma of B-cell type (DLCL). We explored molecular features of DLCL with c-MYC/IGH fusion and the impact of this genetic abnormality on clinical outcome of DLCL.

PATIENTS AND METHODS: A total of 203 cases of non-BL DLCL were studied. Genomic DNA extracted from tumor tissues was subjected to long-distance polymerase chain reaction (LD-PCR) using oligonucleotide primers for exon 2 of c-MYC and for the four constant region genes of IGH.

RESULTS: Twelve cases (5.9%) showed positive amplification; one had a c-MYC/Cµ, nine had a c-MYC/C, and two had a c-MYC/C fusion sequence. Restriction and sequence analysis of the LD-PCR products, ranging from 2.3 to 9.4 kb in size, showed that breakage in the 12 cases occurred within a 1.5-kb region that included exon 1 of c-MYC in combination with breakpoints at the switch regions of IGH (10 of 12). In 10 cases, Myc protein encoded by the fusion genes demonstrated mutations and/or deletions. Six cases had additional molecular lesions in BCL-2 or BCL-6 and/or p53 genes. The age range of the 12 patients was 44 to 86 years, with a median age of 65.5 years. Five patients had stage I/II disease, and seven had stage III/IV disease. Lactate dehydrogenase was elevated in nine of 11 subjects. Seven showed involvement of the gastrointestinal tract. All patients were treated by surgery and/or chemoradiotherapy; six died of the disease within 1 year, resulting in the poorest 1- and 2-year survival rates among DLCL subgroups.

CONCLUSION: The c-MYC/IGH fusion gene of DLCL is identical to that of the sporadic type of BL (sBL). DLCL with c-MYC/IGH shares clinical features with sBL but is characterized further by an older age distribution.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

 
BURKITT’S LYMPHOMA (BL) has been defined on the basis of its characteristic cytomorphology as well as specific translocations involving 8q24, the band in which the c-MYC proto-oncogene is located.¹ However, endemic (eBL) and sporadic (sBL) cases not only exhibit distinct clinicoanatomical features and Epstein-Barr virus (EBV) incidence but also differ in the position of the breakpoints in relation to both c-MYC and immunoglobulin genes (IGs) as partners.^1-3 The positions of the breakpoints on the IGs suggest that sBL tumors arise later in B-cell ontogeny than eBL tumors.^1-3

The International Lymphoma Study Group has proposed the high-grade B-cell lymphoma, Burkitt-like (BLL) category, showing histopathology intermediate between BL and diffuse large-cell lymphoma of B-cell type (DLCL).⁴ BLL was initially determined to lack c-MYC rearrangement but does carry a rearrangement of the BCL-2 gene,⁴ which suggests that BLL is not related to BL but is a subtype of DLCL of follicular center-cell origin. However, a later study indicated that BLL was a cytogenetically heterogeneous disease, and a significant fraction of BLL had chromosome abnormalities affecting the c-MYC locus.⁵ This discrepancy may largely account for the difficulty in distinction between BL, BLL, and DLCL based on their histologic features, even for experienced hematopathologists.⁶

Despite the effort to discriminate between each category of lymphoid diseases by histopathologic and molecular genetic characteristics, t(8;14)(q24;q32) has been observed not only in BL but also in a proportion of cases with intermediate grade DLCL.^7,8 Cytogenetic and clinical analysis of a large series of non-Hodgkin’s lymphoma (NHL) showed that extranodal NHL was more likely to carry t(8;14) and/or c-MYC rearrangement than node-based NHL,⁹ although the presence of t(8;14) did not affect survival of DLCL patients.^7,10 Ladanyi et al¹¹ suggested that a proportion of breakpoints on 8q24 may lie far from c-MYC in DLCL, because the incidence of c-MYC rearrangement as determined by Southern blot analysis was significantly lower than that of t(8;14) by cytogenetic analysis. Thus it is of interest to determine whether cytogenetically identical t(8;14) has the same biologic significance in the development of BL and DLCL.

We have developed a long-distance polymerase chain reaction (LD-PCR) technique to readily detect the c-MYC/immunoglobulin heavy-chain (IGH) fusion gene, which is the molecular equivalent of t(8;14) of sBL (ie, breakages occur at a point close to the c-MYC exon 1 and within switch regions of IGH).^12,13 In this study, we analyzed c-MYC/IGH fusions in a large series of non-BL DLCL. The purpose of this study was to explore (1) the incidence of this genetic abnormality in DLCL, (2) details of the fusions at the nucleotide level, (3) additional molecular lesions in BCL-2, BCL-6, and p53 genes, and (4) the impact of this genetic abnormality resulting from t(8;14) on clinical outcome of patients with DLCL.

	PATIENTS AND METHODS

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

 
Patient Characteristics
From the list of patients with NHL admitted to Kyoto University Hospital (Kyoto, Japan) and related hospitals between 1981 and 1997, we selected a total of 203 patients whose lymphomas showed diffuse proliferation of large cells of B-cell phenotype. In this process, we included the diffuse mixed small- and large-cell (F) as well as the large-cell immunoblastic (H) categories of the Working Formulation (WF).¹⁴ The small noncleaved cell lymphoma, Burkitt’s and non-Burkitt’s type (J), was excluded. Thus the lymphomas of this study corresponded to the diffuse large B-cell lymphoma category of the Revised European-American Lymphoma classification (REAL).⁴ The histologies of relevant cases were reviewed by H.Y. and confirmed to be non-BL DLCL. The lymphoma tissues were subjected to immunohistochemical analysis, surface immunophenotyping, and gene rearrangement analysis using a probe for the J region (JH) of IGH to determine the B-cell origin of lymphoma cells.

Treatments
Of a total of 199 patients in whom treatment details were available, 178 received combination chemotherapy that included doxorubicin or other anthracyclines, whereas nonaggressive regimens were given to six patients. Fifty-one patients were treated with radiotherapy, and primary tumors were surgically resected in 38. Six patients underwent high-dose chemotherapy associated with stem-cell support. Two patients died before initiation of therapy.

Southern Blot Hybridization and DNA Probes
High-molecular-weight genomic DNA extracted from lymphoma specimens was digested with appropriate restriction enzymes and electrophoresed through 0.8% agarose gels. DNA was transferred onto nylon membrane filters (GeneScreen Plus; NEN Research Products, Boston, MA) and hybridized with probes labeled with [alpha phosphorous-32]dCTP (3000 Ci/mmol, Amersham, Tokyo, Japan) using a random labeling system. Hybridization and washing conditions were as recommended by the manufacturer. F372 probe represented the major translocation cluster region of the BCL-6 gene.¹⁵ Probes for the BCL-2 gene were probe b (major breakpoint cluster region [MBR probe]),¹⁶ pFL-2 (minor breakpoint cluster region [mcr probe]),¹⁷ and pB16 (5'-BCL-2 probe).¹⁸

LD-PCR, Cloning into Plasmid, and Nucleotide Sequencing
Designations and sequences of the oligonucleotide primers to detect oncogene/IG fusion genes were described in our previous articles.^12,13 MYC/13 primer, 5'-GTTCATAGGTGATTGCTCAGGACATTTCTGTTAGA-3', was newly designed to be complementary to the 3'-untranslated region of c-MYC exon 3 in the reverse orientation. Each PCR reaction mixture (50 µL) contained 100 ng of genomic DNA, reaction buffer, dNTP mixture, 20 pmol of each primer, and 2.5 U LA Taq DNA polymerase (Takara Shuzo, Kyoto, Japan). PCR cycling variables for long targets were previously described in detail.^12,13,19 PCR products with A-overhangs were ligated into a plasmid vector with 3' T-overhangs at the cloning site (TA Cloning; Invitrogen, San Diego, CA). Transformation and extraction of DNA were performed by established methods. Nucleotide sequencing of the regions of interest was performed with a BigDye Terminator Cycle Sequencing Kit (PE Applied Biosystems, Foster City, CA), and the sequencing reactions were resolved on an automated sequencer (ABI PRISM 310 Genetic Analyzer, PE Applied Biosystems).

EBV Genome
The presence of EBV genome in the tumor was determined by PCR amplification of the region of EBV-encoded small RNA (EBER). The primer sequences were 5'-GTGGTCCGCATGTTTTGATC-3' and 5'-GCAACGGCTGTCCTGTTTGA-3', and the expected size of the PCR product was 190 base pairs (bp).²⁰ DNA from eBL Raji cells was used as a positive amplification control.

PCR-Mediated Single-Strand Conformation Polymorphism (SSCP) Analysis of the p53 Gene and Direct Sequencing
PCR amplification and subsequent SSCP analysis of the p53 gene were carried out as described.²¹ Aliquots of 100 ng of genomic DNA were amplified by PCR in the presence of [alpha phosphorous-32]dCTP, using six sets of primers: two sets for exon 5 and one set for each of exons 6 through 9 of p53. The PCR products were denatured and loaded onto 6% nondenaturing polyacrylamide gels in the presence or absence of 10% glycerol. The sequencing apparatus was cooled continuously with circulating water. After electrophoresis, the gels were dried and exposed to x-ray film at room temperature without intensifying screens. DNA samples that were scored positive by the PCR-SSCP assay were further studied by direct sequencing.

Statistical Methods
Correlations between gene rearrangements and prognostic variables were analyzed with Fisher’s exact test. Survival curves were plotted by the method of Kaplan and Meier,²² and the curves were compared using the log-rank test.²³

	RESULTS

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

 
c-MYC/IGH Fusion Genes Detected by LD-PCR
LD-PCR is a general method that is capable of detecting oncogene/IG fusion sequences up to 30 kb in size.¹³ To amplify the c-MYC/IGH fusion, oligonucleotide primers were designed to be complementary to exon 2 of c-MYC and to the four constant region (CH) genes of IGH, ie, Cµ, C, C, and C.¹² Genomic DNA extracted from the total of 203 DLCL cases were subjected to LD-PCR analysis, and 12 cases (5.9%) showed positive amplification. One case had a fusion with Cµ, nine had a fusion with C, and two had a fusion with C constant region genes (Fig 1 and Table 1). The integrity of high-molecular-weight DNA from each case was verified by amplification of a 10-kb unrelated locus under identical conditions.

View larger version (65K): [in this window] [in a new window]   Fig 1. Ethidium bromide–stained gel electrophoresis of long-distance PCR showing c-MYC/IGH fusion genes. Constant region genes (CH) involved in each fusion are indicated at the bottom. An aliquot of 10 µL was loaded in each lane and electrophoresed through a 0.7% agarose gel. HindIII digested phage DNA (Takara Shuzo, Kyoto, Japan) was used as a molecular size marker (lane M).

Because the rearranged c-MYC gene allele is transcriptionally active under the influence of IGH sequences, it is possible that mutations within the c-MYC gene fused to IGH affect the integrity of Myc protein. To obtain the entire c-MYC coding sequences, we performed another round of LD-PCR using the CH primers in combination with a primer for the 3'-untranslated region of exon 3 (MYC/13). As expected, the LD-PCR products from the 12 cases were 2.3 kb longer than those obtained with the exon 2 primer (data not shown). The coding regions within exons 2 and 3 were amplified by second PCR using the LD-PCR products as templates and cloned into plasmids for nucleotide sequencing. As listed in Table 1, we found point mutations including those leading to amino acid substitutions in nine cases and intragenic deletions of 51 and 204 nucleotides in two cases.

Additional Genetic Lesions in DLCL With the c-MYC/IGH Fusion Gene
Although an oncogene rearrangement resulting from each chromosome translocation in B-cell NHL probably occurs early in the process of tumorigenesis and defines the properties of each tumor, two or more molecular lesions sometimes coexist in a single lymphoma cell.^26,27 We investigated the status of BCL-2 and BCL-6 oncogenes by Southern blot analysis using probes for their breakpoint clusters and examined the relationship between the two genes with IGs by LD-PCR.¹³ Case 472 had two independent lesions in the 5' region of BCL-2. The BCL-2 alleles were fused to the IG and IG light chain genes. The BCL-6 gene was rearranged in three cases (case nos. 854, 893, and 422), and LD-PCR using appropriate primer pairs revealed that two of the three had a fusion with IGH indicative of t(3;14)(q27;q32); thus the two IGH loci were affected by two independent translocations. The partner locus of the remaining case remains to be determined.

It has been shown that a high proportion of BL cell lines as well as fresh tumor materials carry somatic mutations of the p53 tumor suppressor gene.²⁸ We performed PCR-SSCP and direct sequencing analysis of p53 in the 12 DLCL cases. As indicated in Table 1, we found five single-base substitutions within the "hot spot" in four cases, and four of the five mutations caused alteration of coded amino acids. Thus a total of six cases carried additional molecular lesions in BCL-2 or BCL-6 and/or p53.

The presence of the EBV genome was determined by PCR amplification of the region for EBER. No PCR product was obtained in any DNA sample from each case.

Clinical Features of DLCL With the c-MYC/IGH Fusion Gene
Table 2 lists the clinical properties of 12 DLCL patients associated with the c-MYC/IGH fusion gene. The age range of the 12 patients was 44 to 86 years, with a median age of 65.5 years. The male-to-female ratio was 2:1. According to the WF, one had a diffuse mixed small- and large-cell lymphoma, one had a large-cell immunoblastic lymphoma, and the remaining 10 lymphomas corresponded to the diffuse large-cell category. Figure 2 shows representative histologies of the non-BL DLCL cases. Five patients had stage I/II disease and seven had stage III/IV disease. Seven showed involvement of the gastrointestinal tract; three had a tumor of the ileocecal region. Lactate dehydrogenase (LDH) was elevated in nine of 11 subjects for whom data were available. All patients were treated with surgery and/or chemoradiotherapy, and complete remission was achieved in seven patients. However, six died of disease within 1 year. Three patients are currently enjoying long disease-free survival, which could be partly accounted for by the lack of p53 mutation. Case 472 carrying both c-MYC and BCL-2 rearrangements had no history of low-grade lymphoma. None of the patients had evidence for association with AIDS.

View larger version (143K): [in this window] [in a new window]   Fig 2. Representative histologies of non-BL DLCL carrying c-MYC/IGH fusion gene (hematoxylin and eosin stain): (A) case 247 (original magnification, x 100), and (B) case 893 (x 200).

Southern blot analysis of the total 203 DLCL cases revealed that 12 had rearrangement of BCL-2 gene, 43 had rearrangement of BCL-6, and one had both gene rearrangements. To investigate whether DLCL carrying c-MYC/IGH fusion gene constitutes a distinct subtype within the heterogeneous disease of DLCL, we studied the correlations of several of the widely accepted prognostic variables with DLCL carrying either one of the three gene rearrangements and those lacking these molecular lesions. As indicated in Table 3, we compared the c-MYC/IGH fusion (n = 12), c-MYC/IGH-negative (n = 191), BCL-2 rearrangement (n = 12), BCL-6 rearrangement (n = 43) and rearrangement-negative (n = 141) subgroups; the last subgroup was negative for the three molecular abnormalities. Poor performance state, bulky mass, high LDH value, and intermediate/high-risk of international prognostic index tended to be more frequent in the c-MYC/IGH subgroup than in the c-MYC/IGH-negative and rearrangement-negative subgroups, although these differences were not statistically significant.

View larger version (32K): [in this window] [in a new window]   Fig 3. (A) Overall survival of 12 DLCL patients with c-MYC/IGH fusion genes as compared with the remaining 191 DLCL patients. (B) Overall survival of DLCL patients with c-MYC/IGH fusion genes (n = 12), BCL-2 rearrangements (n = 12), BCL-6 rearrangements (n = 43), and those lacking these molecular lesions (n = 141). Five patients carried two of the three gene rearrangements.

	DISCUSSION

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

Cloning and nucleotide sequencing of the c-MYC/IGH fusion genes revealed that breakage of all cases occurred adjacent to exon 1 of c-MYC in combination with breakpoints at the switch regions of IGH in 10 cases. The preferential joining to the downstream CH genes (ie, Cµ v C and C) was described in our previous study on Japanese BL.^12,13 The EBV genome was uniformly absent. Thus it is evident that these DLCL cases carried a t(8;14) identical to that of sBL at the molecular level, and the c-MYC gene was most likely deregulated. Characteristic clinical features included male predominance, high LDH values, and involvement of the gastrointestinal tract at presentation, although we found no statistically significant difference as compared with other DLCL patients. The lymphomas initially responded to chemoradiotherapy; however, duration of the remission was short and six patients died of the disease within 1 year. Nevertheless, the presence of long-term survival in a proportion of the patients suggested that the lymphoma could be cured with appropriate therapy. We therefore concluded that DLCL carrying c-MYC/IGH shares molecular and clinical features with sBL,¹ even though the lymphoma cells lacked characteristic cytomorphology.

Retrospective analysis of DLCL with reference to cytogenetic observations has demonstrated correlations between recurring chromosome abnormalities and survival of patients with DLCL. Negative prognostic indicators included abnormalities of 1q, 2p, 3p, 6q and 17p,^8,10,31,32 although the impact of each aberration on clinical outcome varied considerably among comparable studies. On the other hand, complexity of the karyotype (ie, the number of aberrations) has been reported to be another important prognostic factor.^7,33 t(8;14) was classically described in BL and was associated with poor clinical outcome,³⁴ although later studies demonstrated that survival of patients with t(8;14)-bearing DLCL is not significantly different from that of patients with other DLCL.^7,10 This negative correlation of t(8;14) and prognosis could partly account for the fact that t(8;14) was observed even in low-grade NHL,³⁵ and therefore cytogenetically identical t(8;14) could consist of molecularly heterogeneous populations in terms of oncogenic activation. The present study focusing on the particular molecular lesion resulting from t(8;14), c-MYC/IGH fusion gene, apparently warrants reappraisal of the correlation in DLCL between t(8;14) associated with c-MYC deregulation and rapidly progressive clinical features.

It is of particular interest that nine of the 12 patients were older than 60 years and the median age was comparable to that of other DLCL patients, whereas the majority of patients with sBL are children and young adults.¹ The median age of patients with BLL carrying c-MYC translocation was reported to be 41 years.⁵ These observations raised the possibility that histogenesis of sBL/BLL/DLCL is influenced by age-related factors. Because lymphoma is a disease of the immune system, age-related alterations of immune function could affect the properties of each tumor. Many studies of the relationship between immunity and aging have focused on T-cell function, although impairments in B-cell function have been described.³⁶ Peyer’s patch B-cells from aged mice were reported to respond to a protein kinase C (PKC) activator, phorbol myristate acetate, plus a calcium ionophore, ionomycin, to a lesser degree than those from young mice,³⁷ which suggests that proliferative responsiveness of senescent B-cells was impaired via the calcium-dependent PKC pathway.³⁶ It is possible that lymphoma cells of older patients with intrinsic defects, despite the promotion by c-MYC deregulation, may not be able to proliferate as rapidly as sBL cells in children. On the other hand, BL is characterized by marked macrophage infiltration to ingest apoptotic tumor cells; aging may alter this phagocytic activity.

Cytogenetic and molecular studies of NHL that occurs in the gastrointestinal tract have revealed t(8;14) and/or c-MYC rearrangement in a significant proportion of NHL subtypes that show diffuse aggressive histopathology. These include DLCL, high-grade mucosa-associated lymphoid tissue–derived lymphoma, and small noncleaved-cell lymphoma.^38-40 van Krieken et al^38,39 showed that the former two types of NHL developed in the stomach, whereas the latter lymphoma predominantly occurred in the ileocecal region. In their study, Southern blot analysis, using probes for c-MYC and IGH genes, suggested that t(8;14) of the gastric and ileocecal types differed in the position of breakpoints within IGH (gastric type in the switch region and ileocecal type in the J segments),^39,40 although they did not analyze the breakpoints at the nucleotide level. The present study based on sequencing analysis showed that DLCL can arise in both preferential sites and carry the identical c-MYC/IGH fusion gene. Thus molecular mechanisms responsible for the diverse occurrence of gastrointestinal lymphoma remain to be elucidated.

Chromosomal translocations in B-cell NHL result in juxtaposition of cellular oncogenes to the IGs or equivalent loci, and, in general, coding regions of the oncogenes are not interrupted by the translocation.⁴¹ On the other hand, somatic mutations within the coding region of c-MYC have been observed in eBL, sBL, mouse plasmacytoma, and AIDS-associated lymphomas; some of these mutations occurred independently of t(8;14).^42-44 However, as suggested in one of the reports, previous studies did not prove that these mutations exclusively involved the c-MYC allele that was transcriptionally active.⁴² In the present study, we sequenced the entire c-MYC coding region of the translocated allele that was most likely promoted by the juxtaposed IGH. We are currently investigating whether the positions of mutations and deletions are related to particular domains of the Myc protein and whether these lesions have a pathogenetic role.

It is notable that four cases had additional gene rearrangements associated with other B-cell NHL-specific translocations. In these cases, the BCL-2 or BCL-6 gene was probably deregulated, raising the question of whether these additional molecular lesions can affect malignant phenotypes of lymphoma cells carrying c-MYC/IGH fusion. The sequential activation of c-MYC in t(14;18)/BCL2-carrying lymphoma cells has been described not only in clinical cases²⁶ but also in experimental animals.⁴⁵ It is not clear which of the c-MYC and BCL-6 rearrangements occurs first in a single B cell, because both rearrangements preferentially involve switch regions of IGH. A possible link between c-MYC and BCL-6 is that both gene products are transcription factors that act in the nucleus. Myc is known to play a central role in both the cellular responses to proliferative stimuli and the process of transformation, although diverse functions by connecting with Myc-interacting proteins have been described.^46,47 In contrast, Blc-6 has been reported to be a transcriptional repressor,⁴⁸ which probably needs to be downregulated when B-cells exit from the germinal center.⁴⁹ It remains to be determined whether alteration of target genes of the two gene products confers a synergistic growth advantage on lymphoma cells.

In this study, we identified a subgroup of DLCL carrying the c-MYC/IGH fusion gene that had a molecular structure identical to that of sBL. The disease showed intermediate-grade histopathology but was clinically high grade, and it predominantly occurred in older patients. It is currently unknown whether a very intensive, short-term chemother-apy designed for childhood BL^50,51 is also effective for patients with this particular type of DLCL.

	ACKNOWLEDGMENTS

 
Supported by grants-in-aid from the Japan Ministry of Health and Welfare (nos. 7-29 and 9-10).

We thank Drs M. Abe (Fukushima Medical School, Fukushima), N. Maseki (Saitama Cancer Center, Ina), T. Shimomura, H. Shimada, T. Ichinohe (Shizuoka General Hospital, Shizuoka), F. Matsuyama (Shimada City Hospital, Shimada), K. Kita, H. Miwa, M. Yamaguchi (Mie University, Tsu), Y. Ohno, T. Hayashi, S. Miyanishi (Tenri Hospital, Tenri), T. Suzuki (Shiga Medical Center for Adult Disease, Moriyama), T. Ohno (Otsu Red Cross Hospital, Otsu), H. Fujii (Kyoto First Red Cross Hospital, Kyoto), H. Hayashi, T. Akaogi (Kyoto Second Red Cross Hospital, Kyoto), N. Yasuda (Kyoto City Hospital, Kyoto), K. Miyaoka (Kyoto Min-iren Central Hospital, Kyoto), K. Nasu, N. Tomono, S. Doi (Osaka Red Cross Hospital, Osaka), Y. Konaka (Kitano Hospital, Osaka), H. Sakoda (Sumitomo Hospital, Osaka), T. Kamamoto (Kansai Denryoku Hospital, Osaka), H. Konishi (Kurashiki Central Hospital, Kurashiki), and H. Sawada, Y. Izumi, and K. Fujita (Kokura Memorial Hospital, Kitakyushu) for providing clinical materials and information regarding the patients. We thank Dr A. Tachibana (Radiation Biology Center, Kyoto University, Kyoto, Japan) for technical advice.

	NOTES

 
T.A. is a fellow in cancer research of the Japan Society for the Promotion of Science, Tokyo, Japan.

	REFERENCES

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION REFERENCES

 
1. Magrath IT, Jain V, Jaffe ES: Small noncleaved cell lymphoma, in Knowles DM (ed): Neoplastic Hematopathology. Baltimore, MD,Williams & Wilkins, 1992, pp 749-772

2. Neri A, Barriga F, Knowles DM, et al: Different regions of the immunoglobulin heavy-chain locus are involved in chromosomal translocations in distinct pathogenetic forms of Burkitt lymphoma. Natl Acad Sci U S A 85:2748-2752, 1988[Abstract/Free Full Text]

3. Pelicci P-G, Knowles DM II, Magrath I, et al: Chromosomal breakpoints and structural alterations of the c-myc locus differ in endemic and sporadic forms of Burkitt lymphoma. Proc Natl Acad Sci U S A 83:2984-2988, 1986[Abstract/Free Full Text]

4. Harris NL, Jaffe ES, Stein H, et al: A revised European-American classification of lymphoid neoplasms: A proposal from the International Lymphoma Study Group. Blood 84:1361-1392, 1994[Free Full Text]

5. Macpherson N, Lesack D, Klasa R, et al: Small noncleaved, non-Burkitt’s (Burkitt-like) lymphoma: Cytogenetics predict outcome and reflect clinical presentation. J Clin Oncol 17:1558-1567, 1999[Abstract/Free Full Text]

6. The Non-Hodgkin’s Lymphoma Classification Project: A clinical evaluation of the International Lymphoma Study Group classification of non-Hodgkin’s lymphoma. Blood 89:3908-3918, 1997

7. Offit K, Wong G, Filippa DA, et al: Cytogenetic analysis of 434 consecutively ascertained specimens of non-Hodgkin’s lymphoma: Clinical correlations. Blood 77:1508-1515, 1991[Abstract/Free Full Text]

8. Yunis JJ, Mayer MG, Arnesen MA, et al: bcl-2 and other genomic alterations in the prognosis of large-cell lymphoma. N Engl J Med 320:1047-1054, 1989[Abstract]

9. Kramer MHH, Hermans J, Wijburg E, et al: Clinical relevance of BCL2, BCL6, and MYC rearrangements in diffuse large B-cell lymphoma. Blood 92:3152-3162, 1998[Abstract/Free Full Text]

10. Levine EG, Arthur DC, Frizzera G, et al: Cytogenetic abnormalities predict clinical outcome in non-Hodgkin lymphoma. Ann Int Med 108:14-20, 1988

11. Ladanyi M, Offit K, Jhanwar SC, et al: MYC rearrangement and translocations involving band 8q24 in diffuse large cell lymphomas. Blood 77:1057-1063, 1991[Abstract/Free Full Text]

12. Akasaka T, Muramatsu M, Ohno H, et al: Application of long-distance polymerase chain reaction to detection of junctional sequences created by chromosomal translocation in mature B-cell neoplasms. Blood 88:985-994, 1996[Abstract/Free Full Text]

13. Akasaka T, Akasaka H, Ohno H: Polymerase chain reaction amplification of long DNA targets: Applications to analysis of chromosomal translocations in human B-cell tumors (review). Int J Oncol 12:113-121, 1998[Medline]

14. The Non-Hodgkin’s Lymphoma Pathologic Classification Project: National Cancer Institute sponsored study of classification of non-Hodgkin’s lymphomas: Summary and description of a working formulation for clinical usage. Cancer 49:2112-2135, 1982[Medline]

15. Deweindt C, Kerckaert JP, Tilly H, et al: Cloning of a breakpoint cluster region at band 3q27 involved in human non-Hodgkin’s lymphoma. Genes Chromosomes Cancer 8:149-154, 1993[Medline]

16. Tsujimoto Y, Finger LR, Yunis JJ, et al: Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. Science 226:1097-1099, 1984[Abstract/Free Full Text]

17. Cleary ML, Galili N, Sklar J: Detection of a second t(14;18) breakpoint cluster region in human follicular lymphomas. J Exp Med 164:315-320, 1986[Abstract/Free Full Text]

18. Tsujimito Y, Bashir MM, Givol I, et al: DNA rearrangements in human follicular lymphoma can involve the 5' or the 3' region of the bcl-2 gene. U S A 84:1329-1331, 1987[Abstract/Free Full Text]

19. Akasaka T, Akasaka H, Yonetani N, et al: Refinement of the BCL2/immunoglobulin heavy chain fusion gene in t(14;18)(q32;q21) by polymerase chain reaction amplification for long targets. Genes Chromosomes Cancer 21:17-29, 1998[Medline]

20. Frank D, Cesarman E, Liu YF, et al: Posttransplantation lymphoproliferative disorders frequently contain type A not type B Epstein-Barr virus. Blood 85:1396-1403, 1995[Abstract/Free Full Text]

21. Akasaka T, Muramatsu M, Kadowaki N, et al: p53 mutation in B-cell lymphoid neoplasms with reference to oncogene rearrangements associated with chromosomal translocations. Jpn J Cancer Res 87:930-937, 1996[Medline]

22. Kaplan EL, Meier P: Nonparametric estimation from incomplete observations. J Am Stat Assoc 53:457-481, 1958

23. Peto R, Pike MC, Armitage P, et al: Design and analysis of randomized clinical trials requiring prolonged observation of each patient: II. Analysis and examples. Br J Cancer 35:1-39, 1977[Medline]

24. Hayday AC, Gillies SD, Saito H, et al: Activation of a translocated human c-myc gene by an enhancer in the immunoglobulin heavy-chain locus. Nature 307:334-340, 1984[Medline]

25. Ohno H, Furukawa T, Fukuhara S, et al: Molecular analysis of a chromosomal translocation, t(9;14)(p13;q32), in a diffuse large-cell lymphoma cell line expressing the Ki-1 antigen. U S A 87:628-632, 1990[Abstract/Free Full Text]

26. Thangavelu M, Olopade O, Beckman E, et al: Clinical, morphologic and cytogenetic characteristics of patients with lymphoid malignancies characterized by both t(14;18)(q32;q21) and t(8;14)(q24;q32) or t(8;22)(q24;q11). Genes Chromosomes Cancer 2:147-158, 1990[Medline]

27. Yonetani N, Akasaka T, Akasaka H, et al: Molecular features of a new human lymphoma cell line carrying both BCL2 and BCL6 gene rearrangements. Oncogene 17:971-979, 1998[Medline]

28. Gaidano G, Ballerini P, Gong JZ, et al: p53 mutations in human lymphoid malignancies: Association with Burkitt lymphoma and chronic lymphocytic leukemia. Acad Sci U S A 88:5413-5417, 1991[Abstract/Free Full Text]

29. Akasaka T, Yonetani N, Akasaka H, et al: Detection of t(8;14)(q24;q32) by polymerase chain reaction for long DNA targets: A report of two patients with B-cell non-Hodgkin’s lymphoma. Int J Hematol 67:75-79, 1998[Medline]

30. Offit K, Coco FL, Louie DC, et al: Rearrangement of the bcl-6 gene as a prognostic marker in diffuse large-cell lymphoma. N Engl J Med 331:74-80, 1994[Abstract/Free Full Text]

31. Schouten HC, Sanger WG, Weisenburger DD, et al: Chromosomal abnormalities in untreated patients with non-Hodgkin’s lymphoma: Associations with histology, clinical characteristics, and treatment outcome. Blood 75:1841-1847, 1990[Abstract/Free Full Text]

32. Cabanillas F, Pathak S, Grant G, et al: Refractoriness to chemotherapy and poor survival related to abnormalities of chromosome 17 and 7 in lymphoma. Am J Med 87:167-172, 1989[Medline]

33. Kristoffersson U, Heim S, Mandahl N, et al: Prognostic implication of cytogenetic findings in 106 patients with non-Hodgkin lymphoma. Cancer Genet Cytogenet 25:55-64, 1987[Medline]

34. Fukuhara S, Nasu K, Kita K, et al: Cytogenetic approaches to the clarification of pathogenesis in lymphoid malignancies: Clinicopathologic characterization of 14q+ marker-positive non-T-cell malignancies. Jpn J Clin Oncol 13:461-476, 1983[Abstract/Free Full Text]

35. Ladanyi M, Offit K, Parsa NZ, et al: Follicular lymphoma with t(8;14)(q24;q32): A distinct clinical and molecular subset of t(8;14)-bearing lymphomas. Blood 79:2124-2130, 1992[Abstract/Free Full Text]

36. Doria G, Frasca D: Genes, immunity, and senescence: Looking for a link. Immunol Rev 160:159-170, 1997[Medline]

37. Kawanishi H, Joseph K: Effects of phorbol myristate and ionomycin on in vitro growth of aged Payer’s patch T and B cells. Mech Ageing Dev 65:289-300, 1992[Medline]

38. Ott G, Katzenberger T, Greiner A, et al: The t(11;18)(q21;q21) chromosome translocation is a frequent and specific aberration in low-grade but not high-grade malignant non-Hodgkin’s lymphomas of the mucosa-associated lymphoid (MALT-) type. Cancer Res 57:3944-3948, 1997[Abstract/Free Full Text]

39. van Krieken JHJM, Raffeld M, Raghoebier S, et al: Molecular genetics of gastrointestinal non-Hodgkin’s lymphomas: Unusual prevalence and pattern of c-myc rearrangements in aggressive lymphomas. Blood 76:797-800, 1990[Abstract/Free Full Text]

40. van Krieken JHJM, Medeiros LJ, Pals ST, et al: Diffuse aggressive B-cell lymphomas of the gastrointestinal tract. Am J Clin Pathol 97:170-178, 1992[Medline]

41. Rabbitts TH: Chromosomal translocations in human cancer. Nature 372:143-149, 1994[Medline]

42. Yano T, Sander CA, Clark HM, et al: Clustered mutations in the second exon of the MYC gene in sporadic Burkitt’s lymphoma. Oncogene 8:2741-2748, 1993[Medline]

43. Bhatia K, Huppi K, Splanger G, et al: Point mutations in the c-Myc transactivation domain are common in Burkitt’s lymphoma and mouse plasmacytomas. Nat Genet 5:56-61, 1993[Medline]

44. Bhatia K, Spangler G, Gaidano G, et al: Mutations in the coding region of c-myc occur frequently in acquired immunodeficiency syndrome-associated lymphomas. Blood 84:883-888, 1994[Abstract/Free Full Text]

45. Marin MC, Hsu B, Stephens LC, et al: The functional basis of c-myc and bcl-2 complementation during multistep lymphomagenesis in vivo. Exp Cell Res 217:240-247, 1995[Medline]

46. Schmidt EV: MYC family ties. Nat Genet 14:8-10, 1996[Medline]

47. Gu W, Bhatia K, Magrath IT, et al: Binding and suppression of the Myc transcriptional activation domain by p107. Science 264:251-254, 1994[Abstract/Free Full Text]

48. Albagli O, Dhordain P, Bernardin F, et al: Multiple domains participate in distance-independent LAZ/BCL6-mediated transcriptional repression. Biochem Biophys Res Commun 220:911-915, 1996[Medline]

49. Ye BH, Cattoretti G, Shen Q, et al: The BCL-6 proto-oncogene controls germinal-centre formation and Th2-type inflammation. Nat Genet 16:161-170, 1997[Medline]

50. Magrath I, Adde M, Shad A, et al: Adults and children with small non-cleaved-cell lymphoma have a similar excellent outcome when treated with the same chemotherapy regimen. J Clin Oncol 14:925-934, 1996[Abstract/Free Full Text]

51. Todeschini G, Tecchio C, Degani D, et al: Eighty-one percent event-free survival in advanced Burkitt’s lymphoma/leukemia: No differences in outcome between pediatric and adult patients treated with the same intensive pediatric protocol. Oncol 8:S77–S81, 1997 (suppl 1)

Submitted March 8, 1999; accepted September 30, 1999.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. Nakamura, H. Ye, C. M. Bacon, A. Goatly, H. Liu, L. Kerr, A. H. Banham, B. Streubel, T. Yao, M. Tsuneyoshi, et al. Translocations Involving the Immunoglobulin Heavy Chain Gene Locus Predict Better Survival in Gastric Diffuse Large B-Cell Lymphoma Clin. Cancer Res., May 15, 2008; 14(10): 3002 - 3010. [Abstract] [Full Text] [PDF]

				J. Steere, A. Perl, E. Tomczak, and A. Bagg Large B-Cell Lymphoma Masquerading As Acute Leukemia J. Clin. Oncol., April 20, 2006; 24(12): 1950 - 1951. [Full Text] [PDF]

				G. Moldenhauer, S. W. Popov, B. Wotschke, S. Bruderlein, P. Riedl, N. Fissolo, R. Schirmbeck, O. Ritz, P. Moller, and F. Leithauser AID expression identifies interfollicular large B cells as putative precursors of mature B-cell malignancies Blood, March 15, 2006; 107(6): 2470 - 2473. [Abstract] [Full Text] [PDF]

				I. S. Lossos Molecular Pathogenesis of Diffuse Large B-Cell Lymphoma J. Clin. Oncol., September 10, 2005; 23(26): 6351 - 6357. [Abstract] [Full Text] [PDF]

				J. J. Hoefnagel, R. Dijkman, K. Basso, P. M. Jansen, C. Hallermann, R. Willemze, C. P. Tensen, and M. H. Vermeer Distinct types of primary cutaneous large B-cell lymphoma identified by gene expression profiling Blood, May 1, 2005; 105(9): 3671 - 3678. [Abstract] [Full Text] [PDF]

				A. M. Glas, M. J. Kersten, L. J. M. J. Delahaye, A. T. Witteveen, R. E. Kibbelaar, A. Velds, L. F. A. Wessels, P. Joosten, R. M. Kerkhoven, R. Bernards, et al. Gene expression profiling in follicular lymphoma to assess clinical aggressiveness and to guide the choice of treatment Blood, January 1, 2005; 105(1): 301 - 307. [Abstract] [Full Text] [PDF]

				T. Akasaka, I. S. Lossos, and R. Levy BCL6 gene translocation in follicular lymphoma: a harbinger of eventual transformation to diffuse aggressive lymphoma Blood, August 15, 2003; 102(4): 1443 - 1448. [Abstract] [Full Text] [PDF]

				J. L. Hecht and J. C. Aster Molecular Biology of Burkitt's Lymphoma J. Clin. Oncol., November 1, 2000; 18(21): 3707 - 3721. [Abstract] [Full Text] [PDF]

				T. Akasaka, C. Ueda, M. Kurata, H. Akasaka, H. Yamabe, T. Uchiyama, and H. Ohno Nonimmunoglobulin (non-Ig)/BCL6 gene fusion in diffuse large B-cell lymphoma results in worse prognosis than Ig/BCL6 Blood, October 15, 2000; 96(8): 2907 - 2909. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Akasaka, T. Articles by Ohno, H. Search for Related Content PubMed PubMed Citation Articles by Akasaka, T. Articles by Ohno, H. Social Bookmarking                            What's this?