[CANCER RESEARCH 45, 430-434,
January 1985]
Chromosomal Alterations in Acute Leukemia Patients Studied with Improved
Culture Methods1
Joseph R. Testa,2 Shinichi Misawa, Nobuo Oguma, Keri Van Sloten, and Peter H. Wiernik3
University of Maryland Cancer Center, Baltimore, Maryland 21201
ANLL4 have chromosomal changes. They used a modified culture
ABSTRACT
method involving methotrexate synchronization to obtain more
elongated chromosomes with improved resolution, and they
detected karyotypic changes in everyone of 24 patients with
ANLL. In a subsequent, preliminary study using culture tech
niques we reported chromosomal abnormalities in 25 of 29 (86%)
patients with either ANLL or ALL (18).
We undertook the present study to determine the incidence
and types of chromosomal defects in a larger series of 64 acute
leukemia patients studied with short-term culture methods (both
Cytogenetic studies, using improved short-term culture tech
niques, were performed on 64 patients with acute leukemia to
determine the incidence and kinds of clonal karyotypic changes
detectable with this newer methodology. An adequate number
of analyzable mitoses was obtained from 59 patients. Clonal
chromosomal alterations were found in 88% (52 of 59) of pa
tients, as compared to approximately 50% in previous studies
of acute leukemia in which conventional techniques were used.
From our series, abnormal karyotypes were detected in 37 of 44
(84%) cases with primary acute nonlymphocytic leukemia, all 5
with secondary acute nonlymphocytic leukemia, and all 10 with
acute lymphoblastic leukemia. Among the entire group of pa
tients, several recurrent abnormalities were observed, e.g., -7
with and without methotrexate synchronization). Our results
suggest that most patients with acute leukemia, both ANLL and
ALL, have chromosomal alterations. In some cases, there may
be a subtle rearrangement or a small clone of abnormal cells
only.
in eight cases, +8 in seven cases, t(15;17) in four cases, and
t(8;21) or a variant of this translocation in four cases. In five
patients, the only abnormality was a rather subtle structural
rearrangement (e.g., tiny deletion). Five other patients had clonal
changes which were found in less than 10% of the mitoses
examined in each case. Our results indicate that most patients
with acute leukemia, both acute nonlymphocytic leukemia and
acute lymphoblastic leukemia, have clonal chromosome abnor
malities associated with their disease.
MATERIALS AND METHODS
Sixty-four patients with various types of acute leukemia were exam
ined. Forty-seven of the patients had ANLL, 5 had ANLL secondary to
treatment for a previous malignancy (S-ANLL), and 12 had ALL. Fiftytwo of the 64 patients were studied at diagnosis prior to antileukemic
chemotherapy; the other 12 patients were studied after initiating chem
otherapy and in either initial active disease or in relapse. Fifty-five newly
diagnosed patients were consecutive adults admitted to the University
of Maryland Cancer Center, and one was a child referred for diagnosis;
the remaining 8 adult patients were examined at the time of relapse. The
diagnosis and subclassification of each case was in accordance with the
morphological and cytochemical criteria of the French-American-British
INTRODUCTION
Cooperative Group (2).
In most cases, the cytogenetic specimen was aspirated bone marrow.
Occasionally, an aspirate was unavailable, and a bone core biopsy or
unstimulated peripheral blood was examined in such cases. Specimens
were studied with both the methotrexate cell synchronization technique
(22) and with a modified 24-hr culture method without synchronization.
There have been a considerable number of karyological stud
ies, using chromosome banding techniques, on bone marrow
cells from patients with acute leukemia (for reviews, see Refs.
13, 15, and 16). These earlier studies had suggested that ap
proximately one-half of all patients with acute leukemia have a
Briefly, marrow cells were washed twice with RPM11640 medium, and
then cultured (density: 1 x 106 nucleated cells/ml) in RPM11640 medium
supplemented with 15% fetal bovine serum. Cells were cultured at 37°
normal karyotype. However, most of these investigations were
performed on direct bone marrow preparations, and often the
chromosomes examined were quite contracted and not suitable
for detailed analysis. Several reports have indicated that a short-
in a humidified atmosphere containing 5% CO2. For the
synchronization method, there was an initial incubation
followed by synchronization with 10~7 M methotrexate at
The cells were released with 10~5 M thymidine for 6 hr and
term (24 to 48 hr) culture period can sometimes reveal clonal
chromosomal abnormalities that would not have been detected
if the direct method alone had been used (3, 4,10, 21). Further
more, Yunis et al. (25) suggested that all or most patients with
to Colcemid (0.06 ng/m\) for 10 to 15 min. Cells were swelled with
hypotonie solution (0.075 M KO) at 37°for 20 min and then fixed with
ethanol:acetic acid (3:1). The fixative was changed 4 to 5 times prior to
slide preparation. For unsynchronized cultures, cells were washed and
1Supported in part by the Leukemia Research Foundation, Inc. and by USPHS
Grant 1P50CA-32107 awarded by the National Cancer Institute, Department of
Health and Human Services.
2 Special Fellow of the Leukemia Society of America, Inc. To whom requests for
reprints should be addressed, at Section of Cancer Genetics and Cytogenetics,
University of Maryland Cancer Center, 655 W. Baltimore Street, Baltimore, MD
21201.
'Current address: Albert Einstein College of Medicine, Montefiore Medical
Center, 111 East 210th Street, Bronx, NY 10467.
Received July 23,1984; accepted October 8, 1984.
CANCER RESEARCH
methotrexate
of 3 to 5 hr
37°for 17 hr.
then exposed
* The abbreviations used are: ALL, acute lymphoblastic leukemia; ANLL, primary
acute nonlymphocytic leukemia; S-ANLL, secondary acute nonlymphocytic leuke
mia; L2 and M1 to M6, types of acute lymphoblastic leukemia and acute nonlym
phocytic leukemia, respectively, according to the French-American-British classifi
cation; L2, acute lymphoblastic leukemia with large, heterogeneous blasts; M1,
acute myeloblastic leukemia without maturation; M2, acute myeloblastic leukemia
with maturation; M3, hypergranular promyelocytic leukemia (acute promyelocytic
leukemia); M4, acute myelomonocytic leukemia; M5, acute monocytic leukemia;
M6, erythroleukemia.
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CHROMOSOMAL
ALTERATIONS
IN ACUTE LEUKEMIA
incubated at the same density described above. The cells were cultured
without synchronization for approximately 24 hr and then exposed only
briefly (10 to 15 min) to Colcemid (0.06 ng/m\). Hypotonie swelling,
fixation, and slide preparation were the same as for the methotrexate
synchronization method.
Slides were stained according to our usual laboratory procedures (19).
Mitotic cells were stained conventionally with Giemsa and photographed;
the same cells were then restained and rephotographed with quinacrine
fluorescence to show Q-bands. In certain cases, additional banding
techniques were used. Mitoses were analyzed from the duplicate pho
tographs. Whenever possible, we attempted to analyze at least 10
mitoses from synchronized cultures and 10 from unsynchronized prep
arations for each patient.
Karyotypes were designated according to the recommendations of
the International System for Human Cytogenetic Nomenclature (8, 9).
The observation of a minimum of 2 mitoses with an identical structural
rearrangement or extra chromosome or of 3 cells with the same missing
chromosome was considered evidence for the existence of an abnormal
clone in a patient.
RESULTS
The salient clinical and cytogenetic findings on the 64 patients
studied are listed in Table 1. An adequate number of mitotic cells
for cytogenetic examination was obtained in 59 patients (ANLL,
44; S-ANLL, 5; ALL, 10). In 5 patients (ANLL, 3; ALL, 2), an
analysis of the karyotype was not possible, due to the low
number of mitotic cells and the poor quality of the chromosome
preparations. Overall, clonal karyotypic abnormalities were de
tected in 52 of 59 (88%) évaluablepatients. Of the 44 patients
with primary ANLL, 37 (84%) were cytogenetically abnormal.
Seven ANLL patients had a normal karyotype, including 6 with
M1 type and 1 with M4 type leukemic cells. Abnormal karyotypes
were observed in all 5 patients with S-ANLL and in all 10 with
ALL.
In 5 patients, the only abnormality observed was a subtle
structural rearrangement. Three of these cases (266, 329, 332)
had small interstitial deletions of chromosomes 11,5, and 2,
respectively. A fourth (Case 265) had a defective chromosome
19 which was nearly normal in size but with abnormal, moderate
fluorescence intensity on the short arm (lymphocytes showed a
normal pair 19). The fifth patient (Case 300) had a subtle pericentric inversion, inv(16Xp13q22). One other patient (Case 283)
also showed an inv(16)(p13q22) but, in that case, the abnormality
was accompanied by an easily discernible change, i.e., trisomy
22. Patient 381, a child with Down's syndrome, had tiny intersti
tial deletions of chromosomes 7 and 9 which were accompanied
by an obvious 3; 18 translocation and the constitutional change,
trisomy 21. Examples of several subtle, acquired defects are
shown in Fig. 1.
Five patients (Cases 262, 306, 310, 330, and 346) had clonal
changes which were found in less than 10% of the mitoses
examined in each case. These small clones involved trisomy in
2 patients, structural rearrangement in 2 others, and loss of a
chromosome in the fifth case. In 3 of these cases, the small
clone was recognized only after a considerable number (>50) of
mitoses was fully karyotyped in each case.
Several recurrent abnormalities were oberved, including -7 in
8 patients (ANLL, 4; S-ANLL, 3; ALL, 1), and +8 in 7 (ANLL, 5;
S-ANLL, 1; ALL, 1). Among the structural rearrangements ob
served was a t(15;17) seen in each of the 4 patients with
CANCER
Fig. 1. Qurnacrine-banded chromosomes depicting subtle, acquired structural
rearrangements seen in 3 patients, a, interstitial deletion of chromosome 11,
del(11Xp12p14), in Case 266; b, pericentric inversion of chromosome 16,
inv(16Xp13q22), in Case 300; c and d, interstitial deletions of 7, del(?Xq31.1q31.3),
and 9, del(9)(p22p24 or p21 p23), respectively, in Case 381. Arrows, rearrangement
break points.
hypergranular promyelocytic leukemia (French-American-British
type M3). Four patients had a t(8;21) or a variant of this translocation; 3 had acute myeloblastic leukemia with maturation (M2),
and the fourth had acute myelomonocytic leukemia (M4). Both
patients with an inv(16) had the M4 disease type in association
with the presence of dysplastic bone marrow eosinophils. A
standard Ph1- translocation, t(9;22), was seen in 2 patients with
ALL, type L2. An i(17q) or idic(17q) was seen in 2 other l_2
cases.
Other recurrent structural abnormalities included long arm
deletions of chromosomes 5, 6, and 7. A 5q- was observed in
5 patients (ANLL, 3; S-ANLL, 2). Loss of bands 5q22 to 5q31
was common to 4 of the 5 cases; the fifth had a small deletion,
del(5Xq12q13). A 6q- was found in 3 patients (ANLL, 1; ALL,
2), with loss of 6q21 to 6q23 common to each. A 7q- was seen
in 4 cases (ANLL, 3; S-ANLL, 1). Three of these four patients
had a del(7Xq32) and each had the M1 disease type. The fourth
had a tiny deletion, del(7Xq31.1q31.3), and the diagnosis was
M5.
DISCUSSION
Our results lend support to the proposal by Yunis ef al. (25)
that, with the use of improved culture methods such as the
methotrexate synchronization procedure, most patients with
ANLL can be shown to have clonal karyotypic changes. Further
more, our data suggest that the same may be true for patients
with ALL, as each of the 10 évaluablecases of ALL in this study
had cytogenetic abnormalities. Although results from numerous
RESEARCH VOL. 45 JANUARY 1985
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CHROMOSOMAL
ALTERATIONS
IN ACUTE LEUKEMIA
Table 1
Clinical and cytogenetic findings in patients with acute leukemia
after diag
nosis
of cells
abnor
Laboratory
source"BM8BMBMBMBMBMBM"BMBM"BMBMBMBMBMBM"BM"BMBMBMBMBMBMBMBMBMBMBMBMBMBM
analyzed055
mal"071100248725966710005100953089140100100531004165100140100231001
caserÃ-o.2412422432442482512522542552602612622632652662702712722732752762772782792802812822832862892912952973003033063093103113173203273293
(mos.)1119141.5748575<12.541.55.5735430+173.530+4a1229+1629+14128186128+2319727+174.510.525+18+4825+8.54<
abnormality46,
karyotypic
(43)'28
(16)6(0)29(12)15(0)12(0)28(17)12(5)0(0)11(0)31(5)61
q12?)42,
XX, del(6)(q21q27?), t(15; 17)(q22;
del(8)(p21?),+tdic[dup(3)(p21-^26);
XY, -3, -5, -7, -8, -12, -18,
3acen?
12] (q12?; p13), +A size mar, +1 to
frag93,
mar46,
XXYY, +F size
inv(20)(q13q11or
XY, t(15; 17)(q22; q12?)/46, XY, t(15; 17),
13)47,
3q)46,
XY, +1(1
XX,t(1
XX, t(1 1; ?) (q23; ?)/46, XX, t(1 1; ?), del(6) (q21q25)/46,
+2146,
1; ?), ins(1; ?) (q11; ?)/47, XX, t(11; ?), ins(1; ?),
11)(q13;q23)Sample
XY, t(11;
inadequate46,
+8,del(15)(q13q21?)45,
XY, -16, +der(16)t(8;
(40)23(4)22(11)27
16)(q11 or 12; p13)/47, XY,
-1943,
XY,
q26),del(5)(q13?),
XX, -5, -7, -11, -13, -19, -21, t(2; 10)(q14;
19)(q13;p13)46,+tdic(7; 11)(q11; q13), +der(1 9)t(1 1;
?)46,XX, -19, +der(19)t(19; ?)(p13;
(22)0(0)9(7)43
del(11)(p12p14)Sample
XX,
inadequate45,
frag,del(1)(p31?),
X, -X, -5, -7, +8, -9, -16, -17, -20, -21, +cen
?),+t(21;
t(3; ?) (p25?; ?), +del(5) (q1 1q33)?, t(9; ?) (q12;
mar45, ?)(p13; ?), +A size mar, +C size
-2145,
XY,
(39)42(34)9(5)15(7)0(0)0(0)19(6)10(8)17(9)20(15)19(17)14(13)89
-20,t(9;22)(q34;q11)46,
XX,
t(3;5)(q21;q31)Sample
XY,
inadequateSample
inadequate45,
+mar[inv(20)(p11.2q13.1)?]46,
XY, -7/46, XY, -20,
t(8;21)(q22;q22)48,
XY,
7q),+der(?)
XY, -1 , -2, +5, +1 6, t(8; 10) (q24; q24), del(9) (p22), ¡(1
»2qter)46,
(1pter—»1p11::?),+der dic(1; 2)(1qter—»1p11::2p21—
q12?)47,
XY, t(15; 17)(q22;
inv(16)(p13q22)46,
XX, +22,
del(7)(p15?)46,
XY,
(89)9(8)13(12)30(15)19(16)25
del(9)(q22)46,
XY,
del(6)(q21?q23?)43,
XX,
+der(17)t(5;17)(p13?;
XX, -5, -6, -11, -17, -18, del(7)(q32),
-17,-18,
p12?), +mar/48, XX, -5, +6, +8, +8, +11,
17)46,del(7), +der(17)t(5;
inv(16)(p13q22)46,
XX,
q12?)46,
XY, t(15; 17)(q22;
(24)23(10)23(11)55(10)27(9)32(10)32(9)22(18)48
del(7)(q32)46,
XY,
t(8;21)(q22;q22)47,
XY,
+1845,
XX,
q22)45,
X, -X, t(1 ; 21 ; 8) (q23; q22;
q13)36,
XX, -7/45, XX, -7, t(X; 8)(q26;
(26)34(17)32
(24)62(31)0(0)20(17)31
-17,-20,
X, -X, -2, -3, -5, -7, -9, -12, -13, -15, -16,
inv(3)(q13.3q29)/61,
+21, +B size mar, del(1)(p13.1p22.3),
+del(9)(p12p22),+10,
X, -X, +1p-, +1p-, +4, +6, +8, +8,
+2246,+11, +14, +18, +19, +21, +21, +22,
del(5)(q12q13)46,
XY,
r(18)(p11q23)46,
XY,
del(11)(p11p15)46,
XY,
del(2)(q34q35)Sample
XX,
inadequate46,
p23),+10,
XY, t(13; 19)(q14; p13)/51, X, -Y, +4, +del(6) (p21
del(6)(p21p23),46,
+11,
+13,
+14,
del(5)(q22q31),
(14)28(14)28(15)59
t(8;21)(q22;q22)47,
XX,
+845,
XX,
-747,XX,
+546,
XY,
(22)5(5)18(8)11(0)34(18)7(3)%
t(9;22)(q34;q11)47,
XY,
+646,
XX,
q14)46,
XY, deK6)(p21), del(9)(q12q32), t(10; 14)(q25;
idic(17)(p11)47,
XY, idic(9)(p12)/46, XY, idic(9),
XX, +16
CANCER
RESEARCH
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1985
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CHROMOSOMAL
ALTERATIONS
IN ACUTE LEUKEMIA
Table 1—Continued
diag
abnor
of cells
nosis
Laboratory
mal085093010076100Clonal
analyzed020(3)26(15)28(14)70
abnormality46,
sourceBMBMBMBMBMBMPBNo.
(mos.)21
case
no.374375377378379380381"Age(yr)7064735379321.5SexFFMFMMFDiagnosis"MlM1M4M1M2L2MSSurvivalafter
karyotypic
+1145,
XX, deK7)(q32), t(7; 17)(q22; q23)/47, XX,
+2621
?;14)(6qter-^p21::?::14p11.2-»14qter),
XY, -2, -6, +8, -13, -17, del(5) (q21 q32), +der(6)t(6;
?)(q13.1;45,
+der(20)t(20;
+9<113Sample
(36)30(5)25(14)38
(24)%
+min,+der(8)t(8;
X, -Y, -5, -7, -8, -8, -16, +21, +E size mar, +r,
+21,+der(10)t(10;
?)(q24; ?)/47, XY, -5, -7, +8, -10, -17,
+min46,
17)(p15; q21), +r,
14)47,
XY, t(2; 14)(q21; q32)/49, XY, +13, +19, +21, t(2;
.3),del(9)(p22p24orp21p23)
XX, +21 , t(3; 18) (p23; q1 1.2), del(7) (q31 .1q31
* Diagnosis is primary acute leukemia, unless marked with t, in which case diagnosis is secondary acute leukemia. M1, acute myetoblastic leukemia without maturation;
M2, acute myetoblastic leukemia with maturation; M3, hypergranular promyelocytic leukemia; M4, acute myelomonocytic leukemia; M5, acute monocytic leukemia; M6,
erythroleukemia; L2, acute lymphoblastic leukemia.
b Patients were examined at diagnosis before initiation of chemotherapy, unless marked with * or ", in which case patients were examined after initiation of
chemotherapy and either in initial active disease (*), or in relapse (").
c Total mitotic cells analyzed with banding techniques.
" Indicates percentage of donai abnormal cells.
8 BM, bone marrow; BC, bone core; PB, peripheral blood.
' Numbers in parentheses, subtotal of mitoses analyzed from methotrexate-synchronized cultures.
9 Patient not followed.
" Patient with Down's syndrome (constitutional trisomy 21).
able, and only an unsynchronized culture was set up). We found
no difference in the rates of abnormalities using the 2 types of
culture procedures. Among the cultures with analyzable mitoses,
clonal changes were detected in 48 of 57 (84%) cases without
synchronization as compared to 45 of 54 (83%) with methotrex
ate synchronization. Subtle structural rearrangements were rec
ognized in chromosomes obtained with either method. However,
it should be noted that, since a short exposure to Colcemid and
a considerable number of fixative changes were utilized in both
procedures, it was possible to obtain high-quality, elongated
earlier studies, most of which were done with the standard direct
technique, had suggested that only about one-half of all patients
with either ANLL or ALL have an abnormal karyotype (13, 15,
16), a more recent large compilation (20) revealed a higher
proportion of cases with clonal abnormalities (66%) in ALL which
more closely approximates the incidence reported here.
Yunis ef al. (25) suggested several explanations for the high
incidence of chromosomal alterations in their series of ANLL
patients who were studied with the methotrexate synchroniza
tion method: (a) short-term culture of marrow specimens, either
with or without cell synchronization, may favor division of leukemic cells (3,4,10,21),
so that a 24- to 48-hr incubation period
chromosomes with either culture method.
Yunis et al. (25) proposed that all patients with ANLL may
have a chromosomal defect, whereas Rowley (14) suggested
that at least some of these patients will be found to have a
normal karyotype, even when a large number of cells with
elongated chromosomes are examined. She proposed that leu
kemia in such patients may be related to an alteration in the
control of a normal cellular gene as a result of an insertion of
(submicroscopic) DNA sequences adjacent to it. Our data sug
gest that about 10 to 15% of patients may have a normal
karyotype, even when improved cytogenetic methods are used.
Moreover, Yunis (24) recently reported that further studies in his
laboratory have also revealed a few ANLL patients with a normal
karyotype.
Interesingly, in our series, 6 of 7 patients with a normal
karyotype had the same disease type (M1). Thus, patients with
this diagnosis may be more likely to have normal chromosomes.
Alternatively, such patients may have a submicroscopic alteration
or, possibly, an extremely subtle, consistent microscope change
that can be delineated only with more elongated chromosomes
than we obtained in this study. Moreover, even though a consid
erable number of mitoses (26 to 89; median, 42) was fully
karyotyped in each of our 7 patients who showed no clonal
abnormalities, a small clone may have been overlooked in one
or more cases. Larson ef al. (11) have discussed the limits of
sensitivity for detecting clonal cytogenetic abnormalities. When
only 25 cells are karyotyped, an abnormal cell will be detected
with 95% certainty only if the abnormality is present in at least
can sometimes reveal an abnormal clone that may not be found
in the direct preparation; (o) some of the chromosomal changes
observed by Yunis et al. (25) were subtle rearrangements that
could only be detected with high-resolution banded chromo
somes; (c) with cell synchronization, large numbers of analyzable
mitoses were often available so that small abnormal clones could
be found. In 5 patients from our study, the only karyotypic
abnormality was a subtle structural rearrangement that might
have been overlooked were it not for the improved resolution of
elongated, finely banded chromosomes. With standard cytogenetic techniques, bone marrow chromosomes from leukemia
patients are frequently contracted and poorly banded (16) and,
therefore, subtle changes may be easily missed (22). Five other
patients from our series had small clones (less than 10% of cells
examined). In 3 of these patients, the clonal change was identified
in each case only after a detailed examination of more than 50
mitoses (or about twice the number examined in many cytogenetics laboratories). With improved cytogenetic methods, it is
possible to obtain a greater number of mitotic cells suitable for
detailed analysis and thereby increase the chance of detecting
small clones that might have been overlooked previously.
Overall, results obtained with the improved unsynchronized
and methotrexate-synchronized culture procedures were similar.
Analyzable mitoses were found in 57 of 64 (89%) cultures studied
without cell synchronization and in 54 of 63 (86%) in which
methotrexate was used (in Case 255, too few cells were avail
CANCER
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CHROMOSOMAL
ALTERATIONS
IN ACUTE LEUKEMIA
12% of the total cell population (7). Furthermore, because either
2 or 3 cells with the same chromosomal alteration must be found
to be considered a clone, the leukemic population must therefore
by much greater than 12% of the total cell population to be
recognized (11).
No control sample was available to assess whether some of
the smaller karyotypic changes reported here can occur in the
absence of a known diagnosis of leukemia. In one study of more
than 100 healthy bone marrow transplant donors whose marrow
chromosomes were examined with banding techniques, none
showed a clonal karyotypic abnormality.5 Specimens from these
kemia: a new association. Blood, 67: 994-998, 1983.
2. Bennett, J. M., Catovsky, D., Daniel, M-T., Flandrin, G., Galton, D. A. G.,
Gralnick, H. R., and Sultan, C. [French-American-British (FAB) Cooperative
Group]. Proposals for the classification of the acute leukaemias. Br. J. Haematol., 33: 451-458, 1976.
3. Berger, R., Bemheim, A., and Flandrin, G. Absence d'anomalie chromosomique
individuals were processed by standard cytogenetic procedures,
and similar studies with newer methodology will be needed to
determine the incidence of fine chromosomal defects in hematologically normal marrows.
It is now well recognized that both ANLL and ALL can be
divided into several subcategories, each having a specific chro
mosomal change and distinctive clinicopathological characteris
tics (13, 15, 16, 23). Two samples include the t(15;17) in M3
type ANLL and the t(8;21) which is usually found in the M2
disease type. Our study included examples of many such well
known cytogenetic-clinicopathological
entities. In addition, we
7.
4.
5.
6.
8.
9.
10.
11.
12.
identified 2 patients with an inv(16) who had M4 disease and
dysplastic bone marrow eosinophils. This subtle inversion was
first identified in a patient whose marrow cells were examined
with high-resolution chromosome banding techniques (25). Sub
sequent study of other patients has revealed an interesting
association of this inv(16) and other related abnormalities affect
ing band 16q22 with abnormal eosinophils in ANLL-M4 (1, 5,12,
13.
14.
15.
17).
Improved cytogenetic techniques can be expected to allow for
the identification of further "new" chromosomal rearrangements
16.
17.
associated with particular subgroups of leukemia. Some of these
subtle rearrangements may be quite rare, and the specific cytogenetic-clinicopathological associations will be recognized only
as more patients are studied with newer methodology. Honey
and Shows (6) note that, in ANLL, different chromosomes and,
by implication, different genes are significant in leukemogenesis
at different stages of granulocyte differentiation. Therefore, fur
ther cytogenetic studies with improved techniques will be useful
in uncovering chromosomal sites that may carry genes critical to
the normal control of proliferation and differentiation of particular
bone marrow cells.
18.
19.
20.
21.
22.
23.
REFERENCES
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Chromosomal Alterations in Acute Leukemia Patients Studied
with Improved Culture Methods
Joseph R. Testa, Shinichi Misawa, Nobuo Oguma, et al.
Cancer Res 1985;45:430-434.
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