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MYC Rearrangement and Translocations Involving Band 8q24 in Diffuse
Large Cell Lymphomas
By Marc Ladanyi, Kenneth Offit, Suresh C. Jhanwar, Daniel A. Filippa, and R.S.K. Chaganti
The configuration of the MYC gene in diffuse large cell
lymphomas (DLCL) with translocations involving band 8q24
[t(8q24)] has not been systematically studied. We collected
cytogenetic and clinical data on 171 consecutive cases of
DLCL, including cleaved, noncleaved, and immunoblastic
types, of which 96 had DNA available and 124 had abnormal
karyotypes. The cases with DNA available were evaluated for
MYC rearrangement (MYC-R) by Southern hybridization of
EcoRI-digested tumor DNA using an exon-1 probe, a combination of probe and enzyme known t o detect over 85% of
breaks in sporadic Burkitt’s lymphoma. In cases studied at
diagnosis, MYC-R, t(8;14)(q24;q32), or other t(8q24) were not
prognostically significant. Among the 124 cases with karyotypic abnormalities, seropositivity for human immunodeficiency virus was significantly more common in cases with a
t(8q24) (72%) than in cases without it (9%) (P < .05). Of the
four cases with an MYC -R, two had a t(8;14), one had a
t(7;8;14)(~15;q24;932), and one had a t(8;?)(q24;?) and a
T
HE t(8;14)(q24;q32) translocation, or one of its variants t(2;8)(p12;q24) and t(8;22)(q24;q32), is classically seen in Burkitt’s lymphoma (BL), but is also found in
some diffuse non-Burkitt’s non-Hodgkin’s lymphomas
(NHL).’-4 The t(8;14) translocation has been shown to
juxtapose the MYC oncogene, located at 8q24, to the
immunoglobulin heavy chain (IGH) gene, located at 14q32
resulting in deregulated expression of MYC. Similarly, the
variant translocations involve the light chain IG genes, IGK
at 2p12, and IGL at 22qll.’ Recent studies have shown the
prognostic significance of certain cytogenetic findings within
histologically defined subgroups of NHL.6-8 Because the
clinical significance of t(8;14) in non-Burkitt’s NHL has not
been defined, we determined the incidence and clinical
correlations of a cytogenetic 8q24 translocation break
[t(8q24)] in a consecutive series of 171 diffuse large cell
lymphomas (DLCL), of which 124 had an abnormal karyotype.
In endemic BL (eBL) with t(8;14), the translocation
breaks usually involve the V-DJ regions and sequences far
5’ of MYC, while in sporadic BL (sBL) and acquired
immunodeficiency syndrome (AIDS)-associated BL with
t(8;14), the switch regions of ZGH and MYC sequences in
the first intron-first exon or regions immediately 5’ are
involved?.’ In all variant translocations the breaks are
. ~ the precise mapping of the breaks
located 3‘ o ~ M Y CThus,
in the MYC gene in numerous BL specimens and cell lines
has demonstrated considerable heterogeneity and some
clinical correlations. However, MYC rearrangements have
not been extensively studied in non-Burkitt’s NHL. We
present here the results of breakpoint mapping studies in 15
cases with a t(8q24), taken from the subset of 76 cases of
DLCL where both an abnormal karyotype was documented
and material for molecular studies was available. In addition, we have evaluated the same 15 cases for point
mutations in the PvuII site in the first exon of MYC, because
such mutations at this site have been suggested to contribute to transcriptional deregulation of MYC. lo
Blood, Vol77, No 5 (March 1). 1991: pp 1057-1063
de1(8)(q24). In the three previous cases with translocations
involving 8q24 and 14q32, comigration of the rearranged
MYC band with either the J region or the switch-p region of
the lg heavy chain gene could not be demonstrated, leaving
the 14q32 breakpoint undefined at the molecular level.
Among the remaining 72 cases where both an abnormal
karyotype and molecular data were available, 11 had a
t(8q24). either t(8;14) or t(8;22)(q24;qll), in the absence of
an MYC-R. In these cases, the 8q24 break was presumably
located outside of the EcoRl MYC fragment. All 15 cases with
a t(8q24) were also screened for point mutations in the bull
site in the first exon of MYC; two cases that were not MYC-R
showed loss of this restriction site. These results indicate
that in most DLCL with t(8;14) or other t(8q24). the 8q24
breakpoint lies away from the MYC gene; in a minority of
these cases, point mutations in regulatory noncoding regions
were detected.
o 1991by The American Society of Hematology.
MATERIALS AND METHODS
The present series of 171 cases of DLCL is a subset of the series
of 434 specimens of NHL consecutively ascertained over a 5-year
period (1984 through 1988) at the Memorial Sloan-Kettering
Cancer Center (New York, NY),which has been defined in more
detail elsewhere! In these 171 DLCL, DNA for Southern blotting
analysis was available in 96, and an abnormal karyotype was
obtained in 124; 76 cases had both available, 48 had only abnormal
karyotypes available, 20 had only DNA results available, and 27
cases had neither type of data available. The DLCL were classified
pathologically according to the International Working Formulation,” and included cleaved, noncleaved, and immunoblastic cell
types. There was no histologic component of BL in any of the cases.
The cell surface IG determination and the cytogenetic analysis,
which consisted of short-term culture followed by G- andlor
Q-banding, were performed as previously described.’.’.’’ The karyotypes are described according to ISCN.” Comparison of proportions of cases with specific cytogenetic or molecular genetic
features were performed using the method of inference from
proportion^,'^ and actuarial survival rates were compared using the
method of Kaplan and Meier” and the logrank test for significance.
Immunoperoxidase studies were performed on B3-fixed (B3: 3%
mercuric chloride in 4% formaldehyde) paraffin-embedded sec-
From the Laboratory of Cancer Genetics, Sloan-Kettering Institute
and the Departments of Pathology (Cytogenetics and Surgical Pathology Services) and Medicine, Memorial Hospital, New York, Ny.
Submitted August 20,1990; accepted October 24, 1990.
Supported by Grant Nos. CA-34775 and CA-20194 from the
National Institutes of Health, Bethesda, MD.
Presented in part at the 81st Annual Meeting of the American
Association for Cancer Research in Washington, DC, May 25, 1990.
Address reprint requests to Marc Ladanyi, MD, Cytogenetics Service, Department of Pathology, Memorial Sloan-Kettering Cancer
Center, 1275 YorkAve, New York, NY10021.
The publication costs of this article were defrayed in part by page
charge payment. This article must therefore be hereby marked
“advertisement” in accordance with 18 U.S.C. section 1734 solely to
indicate this fact.
0 I991 by The American Society of Hematology.
0006-4971I91/7705-OO23$3.00l0
1057
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LADANYI ET AL
1058
tions using an avidin-biotin peroxidase complex method.16 The
L26” and UCHL-1’8monoclonal antibodies, specific for B cells and
T cells, respectively, were obtained from Dakopatts (Santa Barbara, CA).
Gene rearrangements were studied by Southern blotting of
DNA extracted from snap-frozen tissue, using an Oncor ProbeTech vacuum blotting device, as previously described: The following combinations of probes and enzymes were used. For the IGH
gene, DNA digested with Hind111 or EcoRI was probed with a
5.5-kb BamHI-Hind111 fragment encompassing the entire J region
(JH), a 1.3-kbEcoRI fragment of the constant-p,region (Ck), and a
2-kb Sst I fragment including all but the 3’ end of the switch-lr. (SF)
region.19For analysis of the configuration of the MYC gene, DNA
digested with EcoRI, HindIII, PstI, PvuII, SmaI, or Xba I restriction enzymes was probed with anXho I-Xba I genomic fragment of
MYC that includes all but the 5’ end of the first exon.” For the
PvuII site in exon-1 of MYC, a double digestion with PvuII and Xho
I restriction enzymes was performed as described:’ followed by
hybridization with the same probe as described above. The ZGH
and MYC gene probes used in this study were gifts from J. Ravetch
and W. Hayward, respectively.
RESULTS
Cytogenetics. Abnormal karyotypes were obtained in
124of 171cases. Of these, 17 (14%) had a classic t(8;14)(q24;
q32) translocation, four (3%) had a t(8;22)(q24;qll) translocation, and two had other 8q24 translocations, including a
t(7;8;14)(plS;q24;q32) in case 408 and a t(8;?)(q24;?) and
de1(8)(q24) in case 230 (Fig 1).In all, 23 cases (19%) had a
translocation involving 8q24.
In addition to t(8q24), other notable karyotypic abnormalities were present in two cases listed in Table 1: case 591
also showed a t(3;22)(q27;qll) and was included in a
previous report of a series of cases with this translocation,2*
while in case 143 a t(8;22)(q24;qll) and a t(14;18)(q32;q21)
were present together. The latter is the only case with
involvement of 18q21 and 8q24 in the same specimen in this
series; in all, 24 of the 124 cases of DLCL with karyotypic
data showed translocations involving 18q21, most often
t(14;18)(q32;q21).
Gene rearrangements. DNA was available for Southern
blot analysis in 96 cases, including 15 cases with a t(8q24)
(Table 1).EcoRI-digested DNA from these specimens was
probed with the MYC exon-1 probe; this combination of
probe and enzyme is known to detect over 85% of breaks in
sBL.” Four cases (4%) displayed an MYC rearrangement
(cases 230,348,408, and 705). Mapping of the breakpoints
was performed by studying the pattern of rearrangements
obtained with additional enzymes, including HindIII, PstI,
PvuII, SmaI, and XbaI (Fig 2). A clustering of breaks was
observed within exon-llintron-1 for cases 348,408, and 705,
and a more downstream break site was detected in case 230;
these results are schematically represented in Fig 3. None
of the 20 cases with molecular data alone were found to be
rearranged for MYC.
The 15 cases with a t(8q24) were evaluated for exon-1
MYC mutations at the PvuII restriction enzyme site (Table
1). In cases 348, 408, and 705, the PvuII fragment was
rearranged; in cases 540 and 591, thePvuII site at the 3’ end
of exon-1 was lost, indicating a small deletion or a point
mutation at this site (Fig 4). The remaining cases showed no
alteration of the PvuII fragments. Hence, 2 of 11 cases
(18%) of DLCL with an 8q24 break remote from the MYC
gene showed point mutation at this site.
Rehybridization of blots with probes for Sp, Cp, JH, and
MYC were performed in cases 348,408, and 705 to detect
comigrating bands. Two rearranged JH bands were present
in each of these cases (data not shown). Case 705 also
showed a rearrangement of Sp, in the presence of a
germline C p band, whereas in cases 348 and 408 both Sp
and C p were in germline configuration (data not shown).
However, in none of the three cases was comigration of
rearranged Sp or JH bands with the rearranged MYC bands
demonstrated.
Zmmunophenoqpes. Surface IG results on fresh tumor
cell suspensions were available in 11of the 15 cases in Table
1. Three cases expressed only ZGM, five cases also expressed other heavy chains, while the remaining three cases
expressed no surface IG. Six cases, including one studied in
frozen section only (case 295), expressed ZGK light chains,
while three expressed ZGL light chains. Case 143 had an
ZGK phenotype in the presence of a t(8;22). The cell
Chromosome
7
a
14
la
22
143
I 230
Patient
Fig 1. Q-banded partial karyotypes of cases 143,
230, and 348. In case 143, the t(8;22)(q24;qll) and the
t(14;18)(q32;q21) are illustrated. In case 408, the
complext(7;8;14)(p15;q24;q32) resulting in translocation of distal 7p material to 8q. of distal 8q to 14q. and
of distal 14q to 7p is shown. In case 230, the
de1(8)(q24) and the t(8;7)(q24;7) are shown. Arrows
indicate breakpoints.
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1059
MYC REARRANGEMENTS IN t(8;14) LYMPHOMAS
Table 1. Clinical, Pathologic, Cytogenetic, and Molecular Data on the 15 DLCL With t(8q24)
UTN
Age/
Sex
Pathology
Site
Surface
IG
MYC
No. of
Pvull Treatment
A. DLCL with t(8q24) and MYC rearrangement
R
G
230 24/M IMB-D
LN None
Cells
Pre
6
14
1
29
21
32
5
5
4
348 291M LNCC-D
SB
None
R
R
Pre
408 35lM IMB-D
705 38/F IMB-D
LN IGG-K
CW ND
R
R
R
R
Post
Pre
2
B. DLCL with t(8:14) withoutMYC rearrangement
70 23lF IMB-D
LN None
G
G
147
60/F
LNCC-D
178 28lM LNCC-D
Post
28
19
Sp
IGG-K
G
G
Post
LI
IGM-L
G
G
Pre
4
22
295 42lM LNCC-D
LN
K
G
G
Post
4
23
534
SP
ND
G
G
Post
1
16
536 31/F LNCC-D
591 37lM IMB-D
SP
LN
IGM-K
IGG-L
G
G
G
D
Pre
Pre
626
60/F
LNCC-D
LN
IGM-L
G
G
Post
19
9
7
18
656
41/F
LNCC-D
BN
IGG
G
G
Pre
3
33
71/F
LCC-D
Karyotype
46,XY
47.X.-Y,+ 1,del(l)(p32),de1(8)(q24),t(2;5)(p23;q35)t(4;?)(pl6:?),t(8:?)
(q24;?),+mar
46,XY
46,XY,t(8; 14)(q24;q32)
47,XY,+5,t(7;8; 14)(pl5;q24;q32)
46,X,der(X)t(X;l)(q26;q21),t(8: 14)(q24:q32),dup(ll)(q13+q23)
47,X,aer(X)t(X;l)(q26;q21),+7,t(8;14)(q24;q32),dup(l l)(q13-+q23)
47,X,der(X)t(X;l)(q26:q21),+7,t(8;14)(q24;q32),dup(l l)(q13+q23)
46,X,der(X)t(X; l)(q26:q21),t(8; 14)(q24;q32),der(4)t(4:?)
(q35:?),dup(l l)(q13+q23)
46,XX,t(8: 14)(q24:q32),d up(11)(q13+q23)
50,XX,+ 11,+12,+ 14,t(1;14)(p31;q32),t(8:14)(q24:q32),der(9)
(qter-cen: :?),der(g)(qter+cen::?), +de1(9)(q22+ter)
48,XX,+1,+7,+9,-20,t(1:6; 1 l)(p31~q23~q13),t(l~13)(q25~q22),t(8;14)
(q24;q32),de1(6)(q21-*ter),del(ll )(pl3+ter)
46,XY
47,XY.- 13,+ 12,t(8;14)(q24;q32),+der(13)t(13;2)(q34;pl5)
46,XX
55-57,XX.6, +7,+22,t(8: 14)(q24:q32),2Xdel(lO)(pl1+ter), der(l3)
t(13;?)(p12;?),+marl,2
46,XX
48,XX. - 15,+6,t(8; 14)(q24:q32).+der(X)t(l;X)(q21;q27),der(2)
(qter+cen::?),+mar
48,XX,+5,t( 1;12)(q12;q13),t(8: 14)(q24:q32),i(6p),+der( 12)t(1; 12)(q12:q13)
47,XY,t(3; 22)(q27;q 11),t(8; 14)(q24;q32)
47,XY,t(3;22)(q27;ql l),t(8;14)(q24:q32),del(2)(q31),+r
51,XX,+3,+ 18,+ 19,t(8; 14)(q24;q32),del(6)(q21+ter),der(3)t( 13;3)
(PI
1;pl l),+del(l)(p22-+ter),+dup(7)(ql l+q36)
98,XX,4n+/- of the clone above
85,XX,+/-,-2,-4,-6,-6,-10,-13,-15,-16,-20,-21,+7,t(8~14)
(q24;q23),+del(l)(pl2-*ter),+der(6)t(6;?:6)(ql3;?;pl3),+marl
C. DLCL with other translocations involving 8q24, without MYC rearrangement
48,X,-X,+7,+8,t(8:22)(q24;ql l),t(14:18)(q32;q21),del(l)(q22+ter),
143 58/F IMB-D
LN IGG-K
G
G
Post
27
+der(l)(qter+q21: :?),der(9)(qter+pl4: :9ql3+ter)
540 74lF IMB-D
LN ND
G
D
Pre
6 46,XX
21
45,X,-X,t(8:22)(q24;qIl)
~~~
~~~~
Abbreviations: UTN. unique tumor number: LN, lymph node: SM, small bowel; CW, chest wall: SP, spleen: LI, liver; BN, bone: IMB-D,
immunoblastic diffuse:LNCC-D, large noncleaved cell diffuse: LCC-D, large cleaved cell diffuse; G, germline; R, rearranged; D, deleted; hull, h u l l
restriction enzyme site in MYC exon-1: Pre, studies performed on pretreatment material; Post, studies performed posttreatment material: ND, not
done.
lineages of the three cases lacking surface immunoglobulin
(cases 70,230, and 348) and of the four cases that were not
studied in cell suspension (cases 295, 534, 540, and 705)
were determined using the L26 and UCHLl monoclonal
antibodies in paraffin sections. Case 230 was a Ki-1 positive
peripheral T-cell lymphoma; its phenotype and genotype
have been reported in detail elsewhere.' The remaining
cases were positive for L26 and negative for UCHL1,
confirming their B-cell lineage.
Clinical correlations. Of the 144 cases with an abnormal
karyotype or DNA available for molecular analysis, 102
were studied before cytotoxic treatment. The median survival of the 14 patients with a t(8q24) or a rearrangement of
MYC was no different from the remaining 88 patients
(P = .23). Serotesting for human immunodeficiency virus
(HIV) was performed in 29 of the 144 cases; of the seven
HIV-positive patients, five had a t(8q24) (71%), including
cases 230,408, and 591 in Table 1, compared with only two
of the remaining 22 patients (9%); this difference was
statistically significant (P < .05).
The salient clinical features of the final study group of 15
cases of DLCL with t(8;14) or other t(8q24) translocations
and DNA available for blotting are summarized in Table 1.
These 15 patients ranged in age from 23 to 76 years, and
there were nine women and six men. The mean age of the
MYC-rearranged group (cases 230, 348, 408, and 705) was
39.5 years compared with 48.4 years for the rest of the
group (P = .lo). The DLCL were classified as large non-
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IADANYI ET AL
1060
408
1
I
E
H
P
S
X
= "&Wb"
705
230
1
I
E
H
P
b
b
b
I
4
* i
;
cleaved cell in seven cases, immunoblastic in seven cases,
and cleaved cell in one case. In none of the cases was a
component of BL observed. Three of the 15 specimens in
Table 1were obtained from extranodal sites: small bowel in
case 348, chest wall in case 705, and bone in case 656. Of the
15 cases in Table 1, cytogenetic and molecular studies were
performed before cytotoxic chemotherapy in eight cases.
Among these, the median survival of the three patients with
rearrangement ofMYC was no different than that of the five
patients with a t(8q24) but witka germline configuration of
MYC(P > SO).
5'
3'
Fig 3. Restriction map of the MYC gene showing the three exons
(hatched boxes, from left t o right), and the regions where the
breakpoints in cases 348,408,705, and 230 are located according t o
the results of the Southern blotting (Fig 2). TheXho I-XbaI fragment of
exon-1 used as the probe is also indicated(open rectangle). Abbreviations: EcoRI, E; Hindlll, H; Pstl, Ps; Arull, Pv;Smal, S; Xhol, Xh; Xba 1,
Xb.
S
X
Fig 2. Southern blot analysis of the MYC gene in
cases 348,408,705, and 230. The DNA samples were
digested with EcoRl (E), Hindlll (H), Pstl (P), Smal (S),
and Xba I(X), yielding germline fragments of approximately 11, 10, 2.7, 2.0, and 6.6 kb, respectively. The
rearranged fragments are indicated by arrowheads.
The sizes of the rearranged fragments are, from left to
right, case 348: 14,10,2.6, and 5.6 kb; case 408: 12,14,
3.5,11, and 12 kb; case 705: 12, 10.5,3.3,10.5, and 9.7
kb; case 230: 11.5 and 8 kb. The comigration of the
rearrangedHindlll fragment with the germline band in
case 348 was confirmed by digestion with C/al (results
not shown). The DNA in case 348 could not be satisfactorily digested with Xba 1.
DISCUSSION
Up to 20% of DLCL show a t(8;14) translocation?"' In
our series, 14% showed this translocation, and another 5%
showed other translocations involving band 8q24. The
incidence of t(8;14) appears to be roughly the same in the
three histologic subgroups of DLCL, namely the cleaved,
noncleaved, and immunoblastic. Recent long-term follow-up studies of the DLCL group have failed to demonstrate any major differences in survival between these
s~btypes?~
Hence, from both the cytogenetic and clinical
viewpoint, grouping the three histologic subtypes for the
purposes of this analysis appears justified.
Two of the cases in Table 1also displayed other characteristic translocations. Case 591 showed a t(3;22)(q27;qll),
a recently identified translocation associated with diffise
NHL." In case 143, the t(8;22) was present in association
with a t(14;18)(q32;q21). Coexistent 18q21 and 8q24 rearrangements, involving BCL2 and MYC, have been described in rare cases of high-grade NHL or acute lymphoblastic leukemia.2s-"Our patient 143 had an immunoblastic
lymphoma and died of disease 7 months after the initial
diagnosis, despite aggressive chemotherapy. However, case
143 is the only case out of 24 with a t(18q21) in our series to
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MYC REARRANGEMENTSIN t(8;14) LYMPHOMAS
10
1.3
-
0.4
-
.
. ~ .-
kb
Fig 4. Southern blot analysis of the MYC gene for point mutation
at the h u l l restriction enzyme site in exon-1 using double digestion
with hull and Xho 1. In cases 540 and 591, the hull site was lost,
presumably by point mutation, producing a new fragment of the
combined size of thetwogermline fragments (1.3kb = 0.9kb + 0.4kb).
Case 348 shows a rearrangement within the bull fragment, producing a new band larger than 1.3 kb. Cases 408 and 705 also showed
rearrangement of the h u l l fragments (not illustrated).The remaining
cases showed no alteration of the h u l l fragments.
also display a t(8q24), suggesting that this is not a common
event in the transformation of low-gradeNHLwith t(18q21).
We found MYC exon-llintron-1 breaks in cases 348,408,
and 705. This region is also typically involved in the t(8;14)
sBL.9s2’With the exception of case 230, the lack of detectable MYC rearrangement in the remaining cases indicates
that the 8q24 break in these cases must lie outside of the
EcoRI fragment containing the MYC gene; this is usually
the case in eBL with t(8;14) and in the variant t(2;8) and
t(8;22) translocations:.” Most breaks in eBL appear to be
located at least 90 kb upstream of the MYC first exon,21
whereas the breaks in the variant translocations occur at
least 10 kb distal to the MYC gene,” beyond the 3’ Hind111
site (Fig 3).
Molecular studies of the MYC gene in cases of non-AIDS
DLCL have been limited. Chenevix-Trench et aI2’ studied
18 DLCL and found a rearrangement of MYC in one case,
located immediately 5’to exon-1; unfortunately, no karyotype data were available on this tumor, or on most of their
other cases. Sozzi et alMreported a case of DLCL with a
t(8;14) and a rearrangement of MYC. Delia et al” studied
the configuration of the MYC gene in five primary cutaneous B-cell DLCL and found it to be germline in all five;
unfortunately, no cytogenetic data were provided. The
presence of Epstein-Barr virus (EBV) within the lymphoma
cells, which is thought to be involved in the pathogenesis of
at least some AIDS-associated NHL, was not studied in the
present series. However, it should be pointed out that no
consistent relationship between EBV infection and the type
1061
of cytogenetic or molecular lesion has so far been shown in
these case^?^"
About 75% of AIDS-associated NHL, both BL and
DLCL, show MYC rearrangement, usually with breaks
similar to those in S B L . ~ ~Accordingly,
”
two of the three
HIV-positive cases in our group of 15 DLCL with t(8q24)
had a rearrangement in MYC. On the other hand, some
AIDS-associated cases show eBL-type breaks in the ZGH
and MYCMgenes. In our series of DLCL, HIV positivitywas
significantly more common in cases with a t(8q24) (71%)
than in cases without it (9%) (P < .05).
Our case 230 was a T-cell Ki-1 positive NHL* and had
cytogenetic involvement of the 8q24 band on both homologues [de1(8)(q24) and t(8;?)(q24;?)], while the two chromosomes 14 appeared normal. One MYC allele, most likely
the one involved in the t(8;?)(q24;?), showed a rearrangement in the 3’ region or immediately downstream; it was
impossible to determine whether the other MYC allele,
presumably the one in the de1(8)(q24) chromosome, was
deleted or germline, due to the high content of nonneoplastic cells in this specimen. However, our data do
allow the exclusion of a rearrangement of this allele.
Translocations involving band 8q24, typically t(8;14)(q24;
qll), have been well described in other T-cell neoplasms;
the break at 8q24 has been localized to the region 3’ of
MYC in several T-cell leukemia cell lines?s36
Possible mechanisms of MYC activation in these translocations include: (1) transcriptional activation by adjacent
ZG enhancers; (2) transcriptional activation by distant ZG
enhancers; (3) mutational inactivation of 5‘ regulatory
sequences by ZG-related hypermutational activity; and (4)
altered half-life and kinetics of a mutated MYC
Whether MYC activation is manifested by absolute overexpression or merely inappropriately sustained expression at
normal levels is still not entirely clear.
In relation to the third mechanism mentioned above,
cases of BL with distant MYC breakpoints, most commonly
endemic cases with t(8;14) or cases with variant translocations, have been found to have a high incidence of point
mutation within and upstream of exon-1 and within intronl?’~’~
The most commonly affected area is the PvuII restriction site at the 3‘ end of exon-1, which is mutated in over
50% of cases, eliminating the restriction site and resulting
in transcriptional deregulation of MYC.
Of the 15 cases
with a t(8q24) in the present series, two cases (540 and 591)
showed evidence of point mutation at the PvuII site; neither
of these two cases was MYC-R. Hence, they appear to be
analogous to eBL with distant MYC breakpoints, although
the incidence of point mutation appears to be significantly
lower (15%). Although the PvuII site is the most common
restriction enzyme site to undergo mutation in this setting,
mutations in exon-1 outside of this site are also numerOUS,Z’*’~ and may be present in at least some other cases in
our series.
The t(8;14) breakpoints within the ZGH locus differs
according to the form of BL? in sBL and AIDS-associated
BL, the breaks usually are in the Sp region, whereas in eBL
they most commonly are located in the JH region. We
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1062
LADANYI ET AL
studied cases 348, 408, and 705 for comigration of rearranged MYC and I G H bands. Case 705 showed rearrangement of both JH and Sp, in the presence of a germline C p
band, indicating that the lymphoma cells had undergone
class switching with deletion of the C p region. In cases 348
and 408, JH was rearranged, but Sp and C p were in a
germline configuration. No surface IG could be demonstrated in case 348; however, monoclonal surface IGG was
present in case 408, suggesting that the class switching that
occurred must have involved the 3’ end of the Sp region, as
previously described in some cases? with deletion of the
remainder of Sp, including the entire region covered by our
Sp probe. In none of the three cases was comigration of the
rearranged S k or JH bands with the rearranged MYC band
demonstrated. Thus, in these cases the 14q32 breakpoint
remains undefined at the molecular level. Occasionally, the
breakpoints in BL have been shown to occur in the D H
region or Sa or ST region^,^^^^' which we did not study.
Cases of BL with the variant translocations t(8;22) and
t(2;8) typically express ZGL and IGK light chains, respectively, because of the sequential rearrangement of the light
chain genes.42Interestingly, however, our case 143 had an
IGKphenotype in the presence of a t(8;22); this exceptional
combination has also been reported in a case of BL.43
Thus, of the 15 DLCL with a t(8q24), three (20%) had
breaks in the 5’ region of MYC, similar to sBL, and another
two cases (13%) showed evidence of point mutation within
exon-1, a regulatoly noncoding region, analogous to some
cases of eBL. One T-cell DLCL had a more 3‘ break in
MYC, as described in other T-cell lymphomas. The molecular events affecting the MYC gene in the other nine cases
(60%)remain to be clarified. It is possible that the t(8;14) in
at least some of these cases is ineffectual in terms of MYC
deregulation, relative to its counterpart in BL, which would
be consistent with its relative lack of impact on survival in
DLCL in the present study, despite the over-representation
of HIV-related cases in the t(8q24) group. In a separate
study, we are currently assessing MYC expression in this
series of DLCL by in situ hybridization. Thus, these studies
show that cases of DLCL with t(8;14) provide another
useful experimental setting for the analysis of the mechanisms of MYC deregulation. Finally, the localization of 8q24
breakpoints at the molecular level in cases of DLCL may
also have future therapeutic implications, because rearrangements within the MYC gene result in abnormal MYC
transcripts that may be targeted in a tumor-specific fashion
by antisense oligonucleotides.@
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1991 77: 1057-1063
MYC rearrangement and translocations involving band 8q24 in diffuse
large cell lymphomas
M Ladanyi, K Offit, SC Jhanwar, DA Filippa and RS Chaganti
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