From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
The
Gene
Located
Diffuse
By Ming-.Sheng
Lee,
at Chromosome
Lymphomas
Mark
Sen Pathak,
B. Blick,
Frederick
B. Hagemeister,
The
karyotypic
abnormality
in 75%
results
in the
which
resides
gene).
65
rearrangement
at
Using
from
the
technique.
mas
transformed
to
large
small
1 8q21
cell
R
cell
cell
lymphomas
of the
Several
been
observed
these somatic
a protooncogene
Burkitt’s
t(8;14)(q24;q32)
B cell
cloned
displayed
had
10
ment
of
13
lymphomas.’3
About
the
75%
display
the
remaining
t(2;8)(pil;q24)
or
the
counterpart
14 band
q32,
chromosomes
chromosome
occur
within
the chromosome
2 band p1 1, and chromosome
band qi 1 where
the immunoglobulin
light chain gene and X light chain gene,
The
still
role
of the
immunoglobulin
genes
In vitro
have
controversial.
oncogene
in
these
studies
situations
develop
a fatal
in these
shown
that
bearing
the
enhancer
and
22
K
overex-
lymphoma.”
77030.
The publication
charge payment.
“advertisement”
indicate this fact.
© 1 987
by Grune
0006-4971/87/7001-001
90
costs ofthis
This article
in accordance
& Stratton.
article were defrayed
in part by page
must therefore
be hereby marked
with 18 U.S.C.
§1734
solely
to
Inc.
1$3.00/0
the
B cell
lines
as a
malignanresiding
at
Recently,
region
were
carrying
the
different
simultaneously
uncultured
in -60%
extent
in all
groups
of
of the
forms
of follicular
rearrangement
of
human
of
malignant
with cytogenetic
analysis,
obtained
directly
from
cells
partictumor
or aspirates.
MATERIALS
AND
METHODS
Procurement
oftissues.
Fresh uncultured
tissues were obtained
from diagnostic
specimens
from 65 patients
at The University
of
Texas MD Anderson
Hospital
and Tumor
Institute
at Houston,
Houston, Texas (32 lymph node biopsies, 25 lymph node aspirates,
3
bone marrow aspirates,
I bone biopsy, I peripheral
blood, 2 pleural
fluid
From the Departments
of Hematology.
Genetics,
Laboratory
Medicine,
Pathology,
and Clinical Immunology,
The University
of
Texas MD Anderson
Hospital
and Tumor Institute
at Houston.
Submitted
September
15, 1986; accepted
February
13, 1987.
Supported
by funds
from
the Hudson
Ard Memorial
Fund,
account No. 1 72! 70.
Address
reprint requests
to Dr Fernando
Cabanillas,
Department
of Hematology,
The University
of Texas MD Anderson
Hospital
and Tumor lnstitute
at Houston,
6723
Bertner Aye, Houston,
TX
no
has
which
bcl-2,
by three
is rearranged
oncogene
the c-myc
oncogene
sequence
fre-
and
Using
these
probes,
these
investigators
that the gene located
at chromosome
I 8 band
is
c-myc
human
cell
translocation
putative
biopsies
gene
involve-
t(14;18)(q32;q21),
oncogene
lymphoid
q21 (the
18q2I
gene)
lymphomas.’3”5
This report
explores
lymphomas
ularly
on
1 8q21
lymphomas,
common
a putative
from
submistudies.
DICIs
18 band
q21, has been postulated.
genomic
DNA
fragments
from
this
investigators.’2’4
demonstrated
this
gene,
reside.6’7
instances
is dysregulated
pressed.8”#{176} Transgenic
mice
coupled
to the
immunoglobulin
quently
heavy
chain
respectively,
of
8
on
most
of
Inc.
of follicular
of the
in
0.03).
=
abnormality,
analogy,
in
no cells
In our
with
(b)
q21
of extranodal
patients
(P
& Stratton,
t(14;i8)(q32;q21)
of
t(8;22)(q24;qI
I) tnanslocation.4
The
common
feature
these
translocations
is the involvement
of chromosome
band q24 where
the c-myc
oncogene
resides.5
Breakpoints
the
human
cloned
with
and
cases
gene.
of the
incidence
in -75%
one
By
chromosome
the
25%
are
cies.
three
kanyotypic
observed
origin.
yielded
1 8q21
18
18 band
rearrangement
compared
by Grune
group
of
and
higher
rearrangement
A similar
of
DLCLs
gene
been
of
translocations
gene.
and
Some
of the
when
1 8q21
S 1987
and
better
a
demonstrated
iden-
cytogenetically
cytogenetically
also
with
that
of unknown
rearrangement
also
(a)
as chromosome
cases
of chromosome
that
a significantly
analysis
situations:
fragment
(44%)
chromosome
We
croscopic
patients
9
cases
metaphase.
(40%)
of
a rearrangement
12 of 1 8 (67%)
(29%)
McLaughlin,
Our
karyotyping
chromosome
4
a 14q+
identifying
analysis
in
cytogenetically
either
in
7
cytogenetics
us with
reciprocal
translocation,
show
donor
genetic
mechanisms
of tukaryotypic
abnormalities
immunoglobulin
lymphomas
the
q21
) translocation
in
involve
an
of
Our
gene
provided
in human
and
1 8q21
t(14;18)(q32;q21
potential
consistent
mutations
(SNCs).
the
DEVELOPMENTS
genetics
have
understanding
morigenesis.
cases
the
2
Peter
Cork,
techniques.
in difficult
band
lympho-
L. Katz,
Ann
cytogenetic
tifying
8 of 20
and
by
helpful
bcl-2,
in 1 8 of 26
Ruth
Goodacre,
in Uncultured
Cabanillas
found
proved
analyzed
follicular
lymphomas,
was
Southern
rearranged
(DLCLs),
of
in which
ECENT
molecular
have
large
rearrangement
cases
was
we
by the
3 of 5 (60%)
lymphomas
noncleaved
detected
(77%)
fragments
J. Butler,
Angela
1 8q21
(the
as probes,
samples
gene
lymphomas,
q21
DNA
q21
lymphoma
The
follicular
oncogene
band
genomic
1 8 band
human
(69%)
diffuse
human
two
18
James
and Fernando
) is reported
translocation
This
of a putative
chromosome
chromosome
uncultured
blot
lymphomas.
M. Trujillo,
S. Velasquez,
U. Gutterman,
t(14;18)(q32;q21
of follicular
Jose
William
Jordan
to occur
18 Band
q21 Is Rearranged
as Well
as Follicular
Lymphomas
aspirates,
and
1 ascitic
fluid
aspirate).
All
were reviewed
by a member of the hematopathology
cytopathology
section in the department
of pathology
be involved
by lymphoma.
Tissue
typing.
All
65 samples
were
classified
of these
section
and
according
samples
and the
proved
to
to the
Working
Formulation
of non-Hodgkin’s
lymphomas
for clinical
usage’6 The determination
of folliculam on diffuse architecture
in the
samples of lymph node aspirates
was based on the histopathology
of
the same lymph node whenever
possible or on a lymph node biopsy
from another site. A subset of patients who displayed
folliculan large
cell lymphoma
(FLCL)
or diffuse large cell lymphoma
(DLCL)
histologically,
but who initially were diagnosed
as having follicular
small cleaved
cell lymphoma
(FSC)
or folliculan
mixed small
cleaved and large cell lymphoma
(FM), were categorized
as transformed
folliculan
lymphoma.
The tumor
immunophenotype
was
determined
by using one or more markers
including
IgG, 1gM, x
light chain, X light chain, BI, iS, Leu-l, Leu-2, Leu-3, Leu-4, Ti),
etc. as described.’7
Blood.
Vol 70. No 1 (July).
1987:
pp 90-95
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
OF THE
REARRANGEMENT
18Q21
GENE
Ea
IN LYMPHOMAS
Es
S
91
SE
Table
1 . Configuration
8;g
‘
I
I
I
V*
i
bcl-2
I
Ill
I
__
Fig 1 .
Partial
restriction
map of chromosome
1 8 band q21 and
probes
used. Top bar represents
the germline
DNA structure
of
chromosome
1 8q21 . Horizontal
lines under chromosome
1 8 mdicate the probes
18q21
and bcl-2
used;
t(14;18)
breakpoint
hot
spot”.
1 . Configuration
of 1 8q21
Human
Gene
Probe
18q21 Probe
Case
No.
Barn
Karyotype
Follicular
small
cleaved
Hind
RI
Pst
Set
Barn
HIAd
RI
R
11
NM
R
12
NM
R
---
13
NM
R
--R
R
14
NM
15
NM
G
G
G--
G
G
G--
16
ND
G
G
G--
G
G
G--A-
R
-
-
R
R
-
A
-
G--
G
ND
G
G
26
ND
G
G
follicular
-
A
R
G
--G
G
G
G
G
G
-
t(14;18)
G
G
53
t(8;14)
G
G
54
t(8;14)
G
55
t(8;22)
R
56
t(8;22)
G
57
ND
G
G
G
-
-
G
-
-
G
-GG-G
G--
G
G
G
-
-
-
R
-
-
-
-
R
-
-
-
G
G
-
-
G
G--
G
G
G
G
G
G
-
G
-
-
R
-
-
-
G
-
-
-
-
G
G
G
G
G
G--
G
G
G--
G
G
-
G
G
-G
GG-
G
G
-G-
GG-
G
G
-G-
G--
G
G
G--
ND
G
lymphoma
iso(18q)
G
G
G
G
-
G
G
--AG
64
46,XY
G
--AG
65
ND
G
G
Hind,
G
GG
-
-
-
-
G
G
G
G
lymphocytic
Abbreviations:
NM,
-
-
Pst,
G
-G-
no metaphases;
Al, EcoAI;
Hindlll;
-
G
Pstl;
Sst,
ND,
SstI;
G
G
G
G
not
done;
G, germline;
G
-
Barn,
-
BamHI;
A, rearrange-
ment.
and large cell
-
-
G
-
A
-
-
A
-
A-AG
-
-
A
-
A-AG
-
G
G
-
-
Preparation
ofDNA
and Southern
blot analysis.
High-mol-wt
DNAs were prepared
by a modification
of the method
described
previously.”
The DNAs
were digested
with various
restriction
endonucleases,
size-fractionated
in 0.8% aganose gels by electnophoresis, and then transferred
onto nitmocellulose
filters as described
by
A
-
A
--
-
A
R--
---
-
A
-A-
Probes were nick-translated
with [a-32P] to a specific activity of 1
to 3 x 10 cpm/,zg of DNA. Hybridization
was carried out at 42#{176}C
overnight
in the presence
of I 0% dextran
sulfate. The filters were
then extensively
washed with 0.1% sodium dodecyl sulfate (SDS)/
0. 1 X sodium chloride
and sodium citrate (SSC) at 65#{176}C.
Autoradiography was done at - 70#{176}C
with intensifying
screens.
Probes used.
Two human genomic
DNA fragments
located at
chromosome
18 band q21, subsequently
called
bcl-2 and 18q2l, were
kindly supplied
by Drs C.M. Croce and Si. Konsmeyer,
respec-
--AG
-
G
-
G-G
G
--AG
G
G-G-
G
G
A
A-AG
lymphoma
27
NM
28
NM
29
ND
A
G
A
30
ND
G
G
GG-
31
ND
G
G
-G
A
A
--
G
-
G
G
52
Small
cell
t(14;18)
G
R
-
--
-
R
-
G
G
cell
R
--
G
-
G
-
--AG
G
-
R
A---
G
G
---
--
-
25
ND
G
-
-
-G
G
A
-
5sf
--G
G
noncleaved
58
G
G
--
R
24
G
Small
T cell
G
-
large
Transformed
-
G
18q2 1 Probe
Hind
RI Pat
-
ND
NM
Follicular
---
NM
10
A
G
63
-
-
R
A
G
62
R
ND
NM
G
-
23
48
NM
NM
A
---G
61
NM
-
R
G
G
9
NM
G
NM
+3
8
t(14;18)
NM
47
59
G
G
21
46
60
-
A
---
GAG
14q+
-
-G
R
G--
7
A
G
R
A
A
G--
A
G
ND
Barn
G
NM
G
A
G
t(14;18)
6q-
ND
-
Sat
--
45
G
R
t(14;18)
44
GG-
G
19
G
G-G
14q+
20
G
G
-
G
-
t(2;4)
G
14q+
R
43
G
6
t(14;18)
G
G
5
18
t(11;14)
50
Pat
G
t(8;14)
42
G
G
R
-
RI
R
41
-
G
t(14;18)
R
-
G
t(14;18)
4
mixed small cleaved
R
G-G
3
t(14;18)
-8,-17,etc.
G
G
17
40
A
A
-
--G
R
t(14;18)
R
G
-
2
-
14q+
G
R
22
Sat
A---
R
R
39
R
t(14;18)
G
Hind
-
1
Follicular
Pst
cell
-
Barn
51
in 65 Uncultured
Lymphomas
bcl-2
Karyotype
in 65 Uncultured
(Cont’d)
Probe
bcl-2
No.
49
Gene
Lymphomas
Case
j
Probes:
Table
of 1 8q21
Human
A
A---
-
G
-AG
G
G
-G-
G
G
-G
G
large cell
32
t(14;18)
A
-
-
33
t(14;18)
A
-
-
-
A
A---
34
14q+
A
-
--
--
A
35
14q+
A
A
36
14q+
A
G
37
14q+
G
G
38
14q+
G
--G
-
tively’’3
(Fig
1).
The
probe
18q21
was
a
1.5-kilobases
(kb)
G
-
A
A
-
A-AG
HindIIl-EcoRl
insert.
The
probe
bcl-2
was a 2.8-kb
EcoRlHindIIl
insert. An immunoglobulin
heavy chain JH gene probe was
provided
by Korsmeyen.#{176} An N-myc
probe was obtained
from
American
Type Culture
Collection
(ATCC,
Rockville,
MD).
Cytogenetic
analysis.
Cytogenetic
analyses were simultaneously
G
G
performed
A
G
previously
descnibed.2”2
Direct
and, if insufficient
metaphases
were then performed.
G
Diffuse
A
A
-
-
-
G-G
G
G
G
--G
G-G
on
tissue
samples
from
5 1 patients
using
techniques
preparations
were performed
first
were observed,
short-term
cultures
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
LEE
92
RESULTS
Of the 65
rearrangement
I ). The
the
rearrangements
rearrangement
cases
or
with
by analysis
(Fig
2A and
and
was
28. Cases
inadequate
case
24, the
probe
l8q21
repeated
enzyme
the
result
the
rearrangements
tion
the
these
mas,
the
lymphomas,
thus, there
analysis.
In
remaining
but
cases
was
restriction
showed
three
within
the
cases
who
that
higher
showed
patients
29,
2).
Twelve
the
I 8q21
the
t(14;18)(q32;q21)
and
ofthese
13 (77%)
20.5-kb
The
three
the
breakpoints
example
of comigration
gene and the immunoglobulin
of the
cases
iH gene. In
ysis
was
confined.
In
cally
C I , D I , E I , and
that a t(14;l8)(q32;q21)
1 1 with cytogenetic
A2,
the filters
B2,
similar
F I ). The
were
C2,
D2,
I 8q2 I probe
rehybridized
E2,
and F2).
size was detected
with
both
the
and the H probe in 28 of the 3 1 samples,
18q21 gene and the JH gene are on the
filters
were
again
washed
to remove
rehybridized
with
the
displayed
indicating
only
that
the
(bcl-2)
samples
fragment,23
detected
with
not the result
a N-myc
1 8q21
probe
of technique
was
then
with a JH probe
A rearranged
as
an
i8q2l
same
the
probes
and
fragment.
The
probe
and
control.
All
the
I8
fragment
9 cases
chromosome
10
gene.
iidicate
that
outde
the
(44%)
that
our analkaryotypi-
unknown
origin,
fragment
that
and the JH probe,
we
hybnidizea
suggesting
translocation
existed.
Three
of the
abnormalities
other than the t(14;18)
or
we demonstrate
a comigrating
of these
on the
have been carefully
of chromosome
chromosome
33
18q2i
which
(cases
patients
structure
remaining
within
chromosome
showed
rearrangement
40, 54, and 55). In none of these
I 4q +
of
suggesting
occurred
of
a comignating
(bcl-2)
probe
51
were
translocation;
no rearrangement
4 of the
14q+
i 8 could
germline
fragments
the iH probe
of the assay.
A.
the
H
internal
a
gene
probes
that
the expected
16-kb
those
rearranged
artifacts
removed,
suggesting
a
could
demonstrate
to both the i8q21
(Fig 3, lanes
fragment
of
(bcl-2)
displayed
gene,
iH
ofthe
chromosome
from
metaphase
rearrangement
the
Of
rearrangement
showed
in
showed
with
showed
restriction
addition
to the 4.3-kb
HindIII
germline
fragment,
each of
the samples
had one on two additional
rearranged
fragments
when analyzed
with the 18q21
probe
(Fig 3, lanes Al, Bi,
and
cells
t(l4;l8)(q32;q21)
with
a significantly
on samples
no
I 8 (67%)
differ-
patients
(P = .03).
disease
attempted
18 cases,
clinical
Companiwith the
revealed
3. The
showed
the
in
lymphomas
were reviewed.
rearrangement
gene
comigration
that
BamHI
20.5-kb
gene (Table
2).
the Ann Arbor
translocation.
13 displayed
restric-
In
of these
gene
cases,
BamHi
lymphocytic
in Table
was
lympho0 of S T cell
small
18q21
with
FSC/FMs
i8q2l
and
of extranodal
BamH
I , indicating
that
of the 4.3-kb
HindIII
I).
the
analysis
(Table
obtained.
The
2,
frequency
SNCs,
no rearrangement
summarized
of
(23
follicular
2 of 7 (29%)
with DLCL
who showed
showed
are
Cytogenetic
enzyme
rearrange(cases
DLCLs,
0 of 2 diffuse
rearrangement
fragment.
restriction
and
lymphomas
transformed
rearrangementofthe
clinical
manifestations
1 2 patients
frag-
that
follicular
3 of 5 (60%)
stage of the 20 patients
son of the eight patients
by using
restriction
31
of 26 (69%)
8 of 20 (40%)
showed
The
rearrangements
HindIII
23 (87%)
in the
l8q21
31
24,
the analysis
HindIII
Of
H1ndIII
correlating
molecular
genetic
analysis
and karyotypes
is shown
in Table
2.
3 FLCLs),
ences
4.3-kb
1).
3 is a representative
rearranged
14, 21,
be detected
of
4.3-kb
(Fig
and
enzymes
cases
which
Eighteen
of
reproduced.
Fig
fragment
fragment
Figure
restriction
only
with
36) were shown by analysis
with
breakpoints
occurred
outside
restriction
I) in 27 of the
were
could
analysis
with
in 23; 20 of these
within
(Fig
or more
breakpoints
I and
rearrangement,
was available
and
the
two
by identification
exceptions
Psi-i,
within
(Table
ment
probes
four
and
clustered
ment
both
rearrangement
molecular
mostly
confirmed
31 showed
gene (Table
I 4, 2 1 , and 28 were small samples;
DNA
to repeat
the Southern
blot
and
The
with probes,
for the I 8q21
were
with
B). The
our results
histopathology
of
samples
we analyzed
of one of the alleles
ET AL
iH
be detected
fragment.
of the
three
The
1 8q2 I
cases
could
karyotypes
reviewed,
with emphasis
18. No abnormalities
in
in any
of these
three.
DISCUSSION
The
were
been
A summary
t(14;i8)(q32;q21)
observed
frequently
chromosomal
in follicular
translocation
lymphomas
has
but
less
B.
I
II
Ill
IV
V
VI
VII
I
II
Ill
IV
V
VI
VII
Fig 2.
Southern
blot hybridization
using probe
1 8q21 . The genomic
DNAs were
extracted
from
cases 27. 1 9. 44. 1 8. 22, 34, and 5. designated
I. II.
Ill, IV. V. VI, and VII. respectively.
(A) Using
enzyme
HindIll.
the 1 8q21 probe detected
a germline band of 4.3 kb. One or two rearranged
bands
(arrows)
were detected
in these samples
except
for case 44 (lane Ill). (B) Using
enzyme
Pat-i.
rearrangements
were also demonstrated
in cases
27. 19, 18, 22. 34, and 5 (lanes I. II, IV, V. VI, and
VII).
The sizes
of the rearranged
bands
were
similar
but not identical.
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
AEARAANGEMENT
OF THE
18o21
GENE
93
IN LYMPHOMAS
A
B
1
2
C
1
2
D
2
1
E
1
2
F
1
2
1
S
kb
I.
.
:IIj
I= g=: #{149}=
:=
Fig 3.
Comigration
of the 1 8q2i gene and immunoglobulin
with a H probe which
recognized
a 1 1 -kb germline
fragment.
probe ,jH (2). Cases 27. 1 9. 1 8, 22, 34. and 5 are designated
demonstrated
to comigrate
with the rearranged
immunoglobulin
frequently
this
DLCL.’3
in
translocation
oncogene
Several
results
bcl-2,
which
sites
breakpoints
base pains
some
resides
segments
of the
The
hybrid
The
The
i8q21
t(14;18)
The
gene component
breakpoint
segment,
restriction
bands
size
enzyme
(5 to 6 kb)
DNA
in
of one
that
of the
gene.
head-to-tail
to
‘ 2.14,2426
probe
the
are usually
enzyme
HindIII.
are
detected
by the
usually
restriction
the
fall within
this
fragment
length
similar
in
cases,
probe
restriction
2.
Correlation
With
of Histology
Rearrangement
i8q2i
to the
18q21
control
probe
i8q2i
sample,
fragment.
The
rangement
by
bcl-2
size
that
in size
from
in these
by
enzyme
detected
than
that
by
of
the
of a rearranged
to the germline
DNA
for detection
the
of the
occurred
and
rear-
breakpoints
on
within
rearranged
band
cases
the
bands,
the other
one
different
in
band.
a 20.5-kb
of
types
bands
is that
“two”
poly-
detected
restriction
instances
detected
the germline
presence
reason
no
)
1 , Psi- I
was
similar
probe
to the germline
analyzed
various
likely
bcl-2
the
18q21
probe
similar
was
and
revealed
In occasional
broader
the
size
most
gene.’2”3
one
of rearbecause
a
subjects
Sst-
I 8q21
using
the germline
suggesting
whose
normal
lymphomas
were
gene
only
possibility
is remote
rearrangement
actually
fragment
was small,
1 , HindIII,
the
45),
>20
of
with rearrangement
enzyme
analyses.
The
than
probe bcl-2
but not by probe
HindIII.
In these instances,
the
and Karyotype
of the
related
chromosome
are com-
from
other
(BamH
14, 2 1 , and
the
interpretation
in the samples
obtained
tumors
enzyme
morphism
We
Table
with
two
since
probe.
polymorphisms
of DNA
of samples
DNA
rearranged
In most
bcl-2
is
I ),24.26
within
the
in
In our series,
most cases
by at least two restriction
assay
could
be performed.
due to DNA
polymorphisms
(cases
HindIII
very
encountered
the amount
patients
J
component.
i8q21
to
5’
When
enzyme
rangement
survey
juxtaposition.
Therefore,
‘ 2.4,2426
I 8q21
3 kb
using
bands
breakpoints
5’ end
heavy-chain
is
is -2
site.
by the
rearranged
to the
problems
rearrangement.
were confirmed
within
450
on chromo-
is 5’ to the JH gene
hot spot on chromosome
which
detected
revealed
HindIII
restriction
enzyme
site (Fig
on chromosome
14q32 usually
occur
breakpoints
J4-J6
close
gene
3’ to the
kb
cases
located
breakpoints
were
immunoglobulin
l8q2I-i
mon
that
of putative
translocation
at the
DNA
sequencing
of
representative
located
showed
heavy chain H gene. The blot shown in Fig 2A was washed
and rehybridized
Autoradiography
shown
by probe 1 8q21 (1 ); autoradiography
shown
by
A. B. C, D, E, and F, respectively.
Rearranged
1 8q21 gene fragments
are
H
gene fragments
( .-).
at 18q21
chromosome
cloning
and
in seven
were
studies
rearrangement
on chromosome
i8q21
(bp) of each other,
and
I4q32
-2.7
previous
in the
The t(i4;I8)
is a unique
molecular
level. Molecular
the crossover
2
BamHl
human
uncultured
segment
lymphomas
of this
and
gene in
demon-
Gene
Karyotype
Histology
t( 1 4; 1 8)
NM
14q +
Others
ND
Total
Table
3.
Clinical
Stages
Lymphoma:
FSC/FM
7/9
1/3
8/9
0/0
1/2
FLCL
1/1
0/0
0/0
0/0
0/2
1/3
Transf. FL
0/0
0/0
2/2
0/0
1/3
3/5
DLCL
2/2
3/6
2/4
1/5
0/3
8/20
SNC
0/1
0/0
0/0
2/4
0/2
2/7
Others
0/0
0/0
0/3
Total
10/13
4/9
12/18
Numerator
,
number
po sitive
for
rearrange
17/23
0/2
0/2
3/11
2/14
ment;
and
do nominat
or, number
small
cleaved
small
metaphases;
cell lymphoma;
cell lymphoma;
formed
no
follicular
noncleaved
FLCL.
follicular
lymphoma;
cell lymphoma.
ND,
FM. follicular
large
DLCL,
not
mixed
done;
small
cell lymphoma;
diffuse
large
FSC,
cleaved
Transf.
cell
lymphoma;
follicular
and large
FL, transSNC,
of
Configuration
18q21
Diffuse
Rearrangement
Large
Nodal
Disease
Gene
Cell
Rearrangement
of the
i8q2i
Gene
I-Ill
IV
Bone
Extranodal
Diseaset
Marrow
Involved
Bone
Marrowt
Spared
Rearrangement
0
4
0
4
Genmline
6
1
4
1
One
NM,
Without
With
With
Stage
analyzed.
Abbreviations:
12 Cases
of 8 Cases
Stage
0/7
31/65
of 20 Patients
Comparison
and
FM
case
showed
Extranodal
tExtranodal
The
histology
sample
disease
.03 by chi-square
tissue,
divergent
in spleen).
that
(including
we
(DLCL
in mesenteric
analyzed
displayed
stage
IV cases)
lymph
node
DLCL.
v nodal
disease
palate,
thyroid,
(P
=
analysis).
sites
pericardium,
of involvement:
lung,
and liver.
base
of tongue,
soft
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
94
LEE ET AL
strated
that
frequently
mas:
a rearrangement
occurs
69%
of follicular
of the
except
histories
they
that
might
of the clinical
manifestations
That
in three
novo
DLCLs,
various
site
exists,
Even
sites
and
BamHI
20.5-kb
has
DLCL
is cumbersome
and
proliferative
follicular
quently
yield
chromosome
20 patients
of the
eight
cases
18q21
gene
was
cases
in
with
of
rangement
there
of
interesting
these
Morphological
subtype
follicular
in patients
t(14;18)
eight
patients
clonality
rearranged
whether
it will
the
part
of
(a)
1 gene
the
have
natural
Does
process
niques.
Molecular
karyotyping
12
The
high
some
rear-
displayed
of
be
followed
from
that
with
the original
many
clone
a newly
to
with
a
transformed
lion?
In our series,
three of five cases
showed
rearrangement
lymphomas
partly answers
the
first
of transformed
follicular
of the I 8q2 1 gene. This
question,
because
the 1 8q2 1 gene can be preserved
determination
of clonality,
however,
follow-up
of the
cases
that
rearrangement
in some cases.
A precise
will require
longitudinal
showed
rearrangement
prior
to
Coexistence
of the
t(14;18)
and
the
t(8;14)
from
with
are
series,
a rearranged
two
lymphomas
SNC
displayed
gene and one carried
the
whereas
the other
carried
t(8;14)(q24;q32)
the t(8;22)(q24;qlI)
tion.
t(8;14)
translocation
(data
not shown).
The
patient
with
rearranged
c-myc
colleagues
postulated
I 8q2
1 gene
results
nant
B cells,
the
oncogene
that
activation
in a clonal
and
acquisition
or t(8;22)j
results
leading
c-myc),
of
expansion
1 8q21
the
high
frequency
in our
studies
samples
were
in the activation
to a higher
grade
and
Tumors
with
rearrangements,
by
cases
that
cytogenetically
We could
in part
finding
be done
in
viable
Furthermore,
aspirates.
due
to the
lymph
The
that
showed
fact
node
that
the
breakpoint
JH
region.
The
showed
yielded
many
rearrangement
of three
cases
of the
18q21
Further
to rule
on chromosome
karyotypes
an intact
of our
aspirates.
of the JH gene.
probe
is required
that
relatively
metaphases
was the demonstration
without
comigration
a Cz, S,s, and VH
however,
it can
analysis.
node
from
gene
with
cases,
in I 2 of I 8
Molecular-
70#{176}C,whereas
-
of 5 1 ) of insufficient
54,
the
tech-
origin.
in that
20 to
-
cytogenetic
obtained
outside
if
cytogenetic
of unknown
of lymph
55)
fredonor
Fur-
requires
a small amount
of DNA.
We
the molecular
genetic
analysis
simply
(I8
and
a very
lymphomas,
gene
of nine
at
for
was
technique
analysis
out the
14 occurred
in each
of these
chromosome
three
1 8, suggesting
a submicroscopic
rearrangement
of the 1 8q2 1 gene. Further
elucidation
of the possible
mechanism
will require
molecular
and
rearrangement
of the 18q21
of B cell lymphomas,
including
types
SNCs.
For
ment
needs
an understanding
to
be
of clonal
studied
Molecular
karyotyping
identifying
clinical
sequencing.
DNA
subsets
evolution,
longitudinally
genetic
is difficult,
analysis
and
of lymphoma
gene occurs
in
DLCLs
and
rearrange-
in
transformed
is useful,
especially
is potentially
with
unique
useful
biological
for
and
behaviors.
rearranged
of low-grade
of a secondary
a
analysis
The
Sometimes
the
is uncertain.
detect
an advantage
(40,
when
had
Pegoraro
the
has
materials
lymphomas.
translocation,
translocaalso
in four
required
from
various
a patient
with
SNC.’4
In our
to
for months
In summary,
translocation
has been observed
in cell lines derived
acute
B cell leukemia27
and a patient
(eg,
cells
cloning
transformation.
t(8;14)
analysis
possibility
of
cells.
follicular
chromosome
An intriguing
clone acquire
a second
of histologic
transforma-
the
the
in cytogenetic
as
internal
Southern
blot analysis
were able to perform
regard
to detect
analysis
proved
very useful
in difficult
We could identify
the donor chromo-
as 18q2l
stored
also
in 10
fell outside
also detect
the t( I 4; 1 8)(q32;q2
I ) translocation
cases that cytogenetically
yielded
no metaphases.
tissues
histoof
able
remain.
viable
such
impossible
a l4q+
genetic
of
used
difficulties
requires
genetic
situations.
could
analysis
fragment.
index,
are
band
not
sequen-
1 3 patients
we
breakpoints
few or no metaphases.
for the 14q+
abnormality
exist,
with
lymphoma
The clinical
history
or does
stage
frequency
to
questions
persist,
in
high
they
of the
cases
be different
Does the original
(b)
that
the
submicroscopic
stage
of the
no rearrangement.
to a more aggressive
Important
be raised:
clone take over?
somatic
mutation
of the
will
is a
1 8q2
from
with follicular
translocation.
was
should
the clinical
in the
in whom
there
transformation
lymphomas.
that
rearrangement.
in view
the
to determine
1 2 patients
no
Arbor
rearrangement
involvement
stage IV presentations
who typically
show
behavior
was
Ann
different
was
extranodal
is
revealed
there
significantly
which
frequency
DLCL
in which
and
in our
genetic
we were
that
significant
low
transfor-
cases
are
that
by molecular
the methodology
improved,
however,
before
analysis
restriction
though
mutations
studies.
revealed
suggests
thermore,
ofthe
gene
abnormality
rearrangement
still
or years
this
(77%).
low-grade
for months
identify
thorough
of each
cytogenetic
t(14;18)(q32;q21)
be follicular
in one
asymptomatic
a
these two somatic
require
longitudinal
with
the
A
from
possibility
undiagnosed
logic
de
biopsies
ie, FM
remote
involved
fide
whether
will also
Correlation
29%
raises
histopathologies
multiple
mation.
Review
the
and
actually
bona
lymphoma
closely
determine
tial events
lympho-
in DLCLs
DLCLs.
and
were
histologies,
The
cases
fragment
B cell
of DLCLs,
gene
into
us
site.
these
18q21
cases
clinical
divergent
another
that
these
for one in which
revealed
gene
human
40%
transformed
convinced
in
of the
that
review
case
this
of
with
of whether
lymphomas
types
lymphomas,
of SNCs.
The rearrangement
question
within
in various
ACKNOWLEDGMENT
malig-
mutation
[eg,
of another
malignancy.27
oncoTo
We wish to thank Dns Croce and Korsmeyer
for kindly supplying
bcl-2 probe and the 18q2l probe. We also wish to thank ioyce
Campbell
for assistance
in the preparation
of this manuscript.
us the
REFERENCES
1. Bboomfield
BA, Gajl-Peczalska
lymphoma.
Cancer
CD,
Arthur
Ki:
DC, Fnizzera
Nonrandom
Res 43:2975,
G, Levine
chnomosomal
1983
2.
EG, Peterson
abnormalities
in
kin’s
Rowley
iD,
lymphoma.
3. Yunis
Fukuhara
Semin
ii,
Oken
Oncol
MM,
S: Chromosome
7:255,
1980
Kaplan
ME,
studies
Ensrud
in non-HodgKM,
Howe
RR,
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
REARRANGEMENT
OF THE
18Q21
Theologides
A: Distinctive
subtypes
of non-Hodgkin’s
GENE
95
IN LYMPHOMAS
chromosomal
lymphomas.
15. Tsujimoto
Y, Cossman
i, iaffe E, Croce
the bcl-2 gene in human
folliculan
lymphoma.
abnormalities
in histologic
N Engl i Med 307:1231,
CM: Involvement
of
Science
228:1440,
I982
198S
Zech L, Haglund
V, Nilsson-Klein
G: Characteristic
chromosomal abnormalities
in biopsies and lymphoid cell lines from patients
with Bunkitt and non-Burkitt
lymphomas.
Int i Cancer
17:47, 1976
5. Dalla-Farera
R, Bregni M, Enikson i, Patterson
D, Gallo RL,
Croce CM: Human
c-myc
oncogene
is located
in the region of
chromosome
8 that is translocated
in Burkitt lymphoma
cells. Proc
16. The Non-Hodgkin’s
Lymphoma
Pathologic
Classification
Project: National
Cancer Institute
sponsored
study of classifications
of non-Hodgkin’s
lymphoma.
Summary
and description
of a working
4.
NatI
Acad
Sci
USA
79:7824,
1982
Croce CM, Shanden
M, Martinis
i, Cicurel L, D’Ancona
GG,
Dolby TW, Kopnowski
H: Chromosomal
location
of the human
immunoglobulin
heavy chain
gene.
Proc Natl
Acad Sci USA
76:3416,
1979
7. McBride
OW, Heiten PA, Hollis OF, Swan D, Otey MC,
Leden P: Chromosomal
location
of human
kappa
and lambda
formulation
6.
immunoglobulin
light
155:1480,
8.
region
A, Drwinga
activation
lymphoma.
9. Croce
CM,
GM,
of the
Proc
NatI
Thierfelder
Nowell
HL,
10.
globulin
i
translocated
Acad
Sci
Enikson
0,
Exp
Med
Croce
CM:
PC,
k locus
to a region
c-myc
transcription.
enhances
CM:
3 of an
Proc
A, Finan
i, Emmanuel
unnearnanged
NatI
i,
unnear-
Acad
B,
of an
immuno-
c-myc
oncogene
Sci
USA
80:7581,
1983
by immunoglobulin
in tnansgenic
12.
Cloning
t(I4;18)
I 3.
i: Amount
by adenovinus
of viral DNA in the
type 2. i Mol Biol
fragments
EM:
Detection
separated
by
gel
of specific
sequences
among
i
Biol
electrophoresis.
Mol
DNA
98:503,
Ravetch
iV,
tion
ofembryonic
and
Sanchez
technique.
Siebenlist
V. Konsmeyer
of the human
0,
rearranged
i and
Escobar
Lancet
5, Waldman
immunoglobulin
2:269,
ii,
locus:
D genes.
Yunis
ii:
T, Leder
Chanacteniza-
Cell
27:583,
A simple
1981
0-banding
1973
Trent
i, Cnickard
K, Gibas
Z, Goodacre
A, Pathak
5,
Sandbeng
AA, Thompson
F, Wang-Peng
i, Wolman
5: Methodologic advances
in the cytogenetic
analysis
of human solid tumors.
Cancer Genet Cytogenet
19:57, 1986
23.
Schwab
M,
Ellison
Bishop
iM:
Enhanced
quent
to amplification
progression
i,
Busch
M,
expression
Roseman
of the
of DNA
of neunoblastoma.
human
may
Proc
W,
gene
Vanmus
N-myc
contribute
Natl
Acad
HE,
conse-
to malignant
Sci
USA
8 1:4940,
1984
I I . Adams
JM, Harris AW, Pinkent CA, Coycoyan
LM, Alexanden WS, Cony 5, Palmitev
RD. Bnister RL: The c-myc
oncogene
driven
1982
22.
by translocation
of a C
Natl
Acad Sci USA
Translocation
49:2112,
1973
P: Structure
in
K, Finan
Cancer
U, Sambrook
transformed
19. Southern
21.
of an
usage.
1975
1983
activation
K, an-Rushdi
Croce
oncogene
80:820,
i, Nishikurak
Transcriptional
J, Nishikura
Nowell
PC,
c-myc
USA
W, Enikson
PC:
Nowell
ranged and untnanslocated
c-myc
oncogene
locus in Burkitt’s
lymphoma
cells. Proc
80:6922,
1983
Lenoir
genes.
Pattensson
of cells
73:125-130,
20.
i, ar-Rushdi
Transcriptional
Lenoir
constant
18.
genome
1982
Enikson
Bunkitt
chain
for clinical
Hsu SM, Raine L, Fanger H: Use of avidin-biotin-peroxidase
complex
(ABC)
in immunopenoxidase
techniques.
J Histochem
Cytochem
29:577, 1981
17.
Tsujimoto
mice.
enhancers
Nature
Y,
induces
318:533,
Finger
LR,
lymphoid
malignancy
1985
Yunis
i,
Nowell
PC,
Croce
CM:
of the chromosome
breakpoint
of neoplastic
B cells with the
chromosome
translocation.
Science 226:1097,
1984
Bakhshi
A, iensen
iP,
Goldman
P. Wright
ii,
McBride
OW,
Epstein AL, Korsmeyer
Si: Cloning the chromosomal
breakpoint
of
t(l4;18)
human lymphoma:
clustering
around iH on chromosome
14
and near a transcriptional
unit on 18. Cell 41:899, 1985
14. Cleary
ML, Sklan i: Nucleotide
sequence
of a t(14;l8)
chromosomal
breakpoint
in follicular
lymphoma
and demonstration
of a breakpoint-cluster
region near a transcniptionally
active locus on
chromosome
18. Proc Natl Acad Sci USA 82:7439,
1985
24.
Tsujimoto
Y, Gorham
t(14;18)
chromosome
result from mistakes
25.
analysis
Cleary
26.
scripts
follicular
ML,
Smith
of cDNAs
transcript
1986
for
resulting
Tsujimoto
and
i, Cossman
translocations
in VDJ joining.
from
Y,
protein
lymphoma.
SD,
bcl-2
Croce
Sklan
and
the
i:
CM:
The
Acad
and
tnanslocation.
the
structural
bcl-2/immunoglobulin
Analysis
of bcl-2,
Proc NatI
E, Croce
Cloning
a hybrid
t(14;l8)
CM:
products
i, iaffe
involved
in B-cell neoplasms
Science 229: 1390, 1985
of
the
Cell
structure,
gene
involved
Sci USA
83:5214,
47:19,
tran-
in human
1986
Pegoraro
L, Palumbo
A, Enikson i, Falda M, Giovanazzo
B,
Emanuel
BS, Rovera 0, Nowell PC, Croce CM: A 14;18 and 8;14
chromosome
translocation
in a cell line derived from an acute B-cell
27.
leukemia.
Proc
Natl
Acad
Sci
USA
8 1 :7 1 66,
1984
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
1987 70: 90-95
The gene located at chromosome 18 band q21 is rearranged in uncultured
diffuse lymphomas as well as follicular lymphomas
MS Lee, MB Blick, S Pathak, JM Trujillo, JJ Butler, RL Katz, P McLaughlin, FB Hagemeister, WS
Velasquez and A Goodacre
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