GENES, CHROMOSOMES & CANCER 29:89 –95 (2000)

PERSPECTIVE ARTICLE

Mechanism and Relevance of Ploidy in

Neuroblastoma
Yasuhiko Kaneko1* and Alfred G. Knudson2
1
Department of Cancer Chemotherapy, Saitama Cancer Center Hospital, Ina, Saitama, Japan
2
Fox Chase Cancer Center, Philadelphia, Pennsylvania

Neuroblastoma has a broad spectrum of clinical behavior, ranging from spontaneous regression to dissemination and fatality.
The heterogeneity that has long puzzled many investigators has been shown by more recent studies to be closely correlated
with various clinical and genetic factors. Tumor cell ploidy is one of the factors; diploid and near-triploid neuroblastomas show
poor and excellent clinical outcomes, respectively. We offer a hypothesis that explains how the ploidy state of the tumor plays
a fundamental role in this heterogeneity, and why various prognostic factors are correlated with each other. This hypothesis
may be applicable to tumors other than neuroblastoma. © 2000 Wiley-Liss, Inc.

Neuroblastoma is a notorious childhood tumor unfavorable, advanced-stage disease in older chil-

because a high fraction of tumors occurring in pa-
tients older than one year is disseminated (Stage 4)
at the time of diagnosis and incurable (Brodeur and
Castleberry, 1997; Matthay et al., 1999). The con-
trast between this very malignant subgroup and the
group of more benign stages (1, 2, and 4S) is so
extreme that the two groups of tumors seem like
different diseases. This raises questions about
whether there are fundamental genetic differences
between the two groups, and whether the Stage 4
tumors begin as such, or whether they have pro-
gressed from the other stages.
It has been known for some time that most Stage
1, 2, and 4S tumors are near-triploid in their chro-
mosomal constitution, whereas the majority of
Stage 4 tumors are diploid or tetraploid (Kaneko et
al., 1987, 1990; Christiansen and Lampert, 1988;
Hayashi et al., 1989; Ambros et al., 1995). Two
recent sets of observations that bear on ploidy and
on the above questions have stimulated us to offer
tentative answers, an explanatory hypothesis, and a
set of predictions.
Results of Mass Screening Program
One set of observations concerns a mass screen-
ing (MS) program for infants that has been per-
formed in Japan and in Quebec province in Canada
(Sawada et al., 1984; Woods et al., 1996). Screening
has consistently led to an increased detection of
neuroblastoma, mostly of low stages; 75% of the
Japanese and 67% of the Canadian infants identi-
fied by screening had Stages 1, 2, and 4S tumors
(Woods et al., 1996; Committee on mass screening
program for neuroblastoma in Japan, 1999). Screen-
ing, however, has not resulted in a decrease of
dren (Yamamoto et al., 1995; Woods et al., 1996).
These findings strongly suggest that screening has
detected a large number of neuroblastomas that
would have regressed spontaneously, and that this
group of neuroblastomas may have special genetic
characteristics.
Abnormalities of Ploidy
A second set of observations was obtained by flow
cytometric and cytogenetic analysis of neuroblas-
toma. In a flow cytometric study by the Pediatric
Oncology Group (POG), the clinical relevance of tu-
mor cell ploidy was examined in 298 patients (Look
et al., 1991). Among infants, hyperdiploidy, mostly in
the near-triploid range, was closely associated with
⬎90% long-term survival, whereas diploidy invariably
predicted early treatment failure. In addition, in chil-
dren 12–24 months of age, diploidy predicted early
failure with chemotherapy, whereas half of the chil-
dren with hyperdiploidy achieved long-term disease-
free survival. These results underscore even more
strongly the idea that hyperdiploidy is correlated with
a benign state. There was no relationship, however,
between ploidy and treatment outcome in children
older than 24 months with widely disseminated dis-
ease, although the ratio of hyperdiploid to diploid
cases was greatly reduced.
Supported by: Ministry of Health and Welfare of Japan (for
Second-Term Comprehensive 10-Year Strategy for Cancer Control).
*Correspondence to: Dr. Yasuhiko Kaneko, Saitama Cancer Cen-
ter, Hospital, Ina, Saitama 362-0806, Japan.
E-mail: kaneko@cancer-c.pref.saitama.jp
Received 3 February 2000; Accepted 22 April 2000

© 2000 Wiley-Liss, Inc.

90 KANEKO AND KNUDSON

TABLE 1. Relationship between the Modal Chromosome Number and the Number of Chromosomes 1 Determined by the
D1Z1 FISH Signal Number in 151 Neuroblastomas*
Modal chromosome number (chromosomal ploidy range)
Number of
chromosomes 1 Number of 44–57 58–80 81–103 104–115
by FISHa tumors Near-diploid Near-triploid Near-tetraploid Near-pentaploid

2 34 31 2 1 0
3 57 9 45 2 1
3/4b 38 1 16 21 0
4 9 0 4 5 0
5 13 0 3 1 9
*The data are based on the study published by Kaneko et al., 1999.
a
Determined by the D1Z1 signal number, e.g., 2 means more than 50% of 100 cells examined showed 2 D1Z1 FISH signals.
b
Indicates a mixed population of cells with trisomy 1 and cells with tetrasomy 1, and each group of cells was found in 25% or more of 100 cells examined.
The number of chromosomes 1 correlated with the ploidy number (P ⬍ 0.0001).

In a cytogenetic study of 68 neuroblastomas, The karyotype was successfully examined in 7 of

Kaneko et al. (1990) found 17 near-diploid, 35
near-triploid, 10 near-tetraploid, and 6 near-penta-
ploid tumors. The near-diploid and near-tetraploid
tumors were usually characterized by a few struc-
tural abnormalities, most commonly involving 1p,
and by the frequent presence of double minute
chromosomes (dmin) or homogeneously staining
regions (hsr), and were mainly found in children
older than one year (Kaneko, 1987, 1990). The
near-triploid tumors were characterized by three
almost complete haploid sets of chromosomes, with
few structural abnormalities, and were mostly
found in infants less than 12 months of age. Pa-
tients with near-pentaploid tumors, like those with
near-triploid tumors, had favorable clinical and bi-
ological prognostic factors and high survival rates.
In contrast, unfavorable prognostic factors were ob-
served in patients who had either near-diploid or
near-tetraploid tumors, and this subset of patients
had a poor outcome.
More recently, Kaneko et al. (1999) have per-
formed metaphase cytogenetic or interphase
2-color fluorescence in situ hybridization (FISH)
analyses using subterminal 1p (D1Z2) and pericen-
tromeric 1q (D1Z1) probes in 246 patients with
neuroblastoma. Chromosome numbers were deter-
mined in 151 of 170 tumors in which the constitu-
tion of chromosome 1 was successfully examined
by FISH. The relationship between the modal
chromosome number and the number of chromo-
somes 1 determined by FISH is shown in Table 1.
The number of chromosomes 1 correlated with the
ploidy number (P ⬍ 0.0001), and thus, tumors with
disomy, trisomy, tetrasomy, and pentasomy 1 may
be considered as tumors with diploidy, triploidy,
tetraploidy, and pentaploidy, respectively.
9 triploid or tetraploid tumors with tetrasomy 1,
and 2 of 3 triploid or tetraploid tumors with disomy
1 (Tables 1 and 2). All 9 patients were ⱖ15 months
old, had advanced stage tumors, and 8 of the 9 had
a poor outcome. One tumor (No. 1185) had near-
diploid cells with one 1p- chromosome and near-
tetraploid cells with 2 of the same 1p- chromo-
somes, suggesting that the near-tetraploid cells
may have arisen from the near-diploid cells, prob-
ably through endomitosis or endoreduplication. All
9 tumors but one (No. 1193) had 2 normal chromo-
somes 1 and 2 identical 1p- chromosomes (7 tu-
mors), or 2 normal chromosomes 1 and 2 identical
derivative chromosomes containing partial 1q (one
tumor; No. 694). These findings suggest that the
near-triploid cells with tetrasomy 1 may have arisen
from near-tetraploid cells, followed by chromo-
some loss. The sequence of tetraploidization with
chromosome loss has been proposed as a model for
the genetic evolution of human solid tumors
(Shackney et al., 1989).
As we mentioned earlier, the POG study using
flow cytometry showed a favorable outcome in in-
fants with hyperdiploid tumors, but an unfavorable
outcome in older children with hyperdiploid tu-
mors (Look et al., 1991). The discrepancy would be
resolved if hyperdiploid tumors that occur in in-
fants were “de novo” triploid tumors, and if most
hyperdiploid tumors in older children were tet-
raploid tumors or “secondary” triploid tumors de-
rived from the sequence of tetraploidization with
chromosome loss. We will describe later how the
“de novo” triploid tumors arise.
On the basis of the findings for chromosome 1
determined by 2-color FISH, the tumors were clas-
sified into four groups: a Dis1Norm1p group (di-
 PLOIDY AND NEUROBLASTOMA 91
TABLE 2. Representative Karyotypes of 9 Neuroblastomas With Disomy 1 or Tetrasomy 1 Determined by FISH and With
Triploid or Tetraploid Chromosome Number*

Number of
chromosomes 1
Patient Age by FISH Stage Karyotypea
369 2y5m 4 3 67–74(4n),XX,⫺X,⫺X,del(1)(p13)⫻2,⫺2,⫺5,⫺6,add(7)(q36)⫻2,⫺8,⫺8,
⫺9,⫺10,⫺10,⫺12,⫺14,⫺14,⫺16,⫺17,⫹18,⫺19,⫺19,⫺20,⫺20,add(22)
(q13)⫻2,dmin [cp12]
460 2y1m 4 4 73–78(4n),XXX,⫺X,der(1)inv(1)(p11q24)del(1)(p34)⫻2,hsr(1)(q42),⫺3,
⫺3,⫺4,⫺6,del(7)(p11),⫺7,⫺8,⫺9,⫺9,add(10)(p11),⫺10,⫺10,⫺11,⫺12,
add(14)(q24)⫻2,⫺15,⫺17,⫺19,⫺19,⫺21,del(22)(q13),⫹4mar
694 2y11m 2 4 69(4n),XX,⫺X,⫺X,⫺1,⫺1,⫺2,⫺3,⫺3,⫺4,⫺5,⫺8,⫺10,⫺11,⫺13,⫺13,
⫺14,⫺15,⫺15,der(16)t(1;16)(q21;q22)⫻2,⫺19,⫺21,⫺21,⫺22,⫺22,⫺22
858 6m 4 4 77–78(4n),XXX,⫺X,add(1)(p13)⫻2,⫺2,⫺3,⫺4,del(4)(p14),der(4)add(4)
(p16)add(4)(q31)⫻2,add(8)(q24)⫻2,⫺9,⫺10,add(11)(q25),⫺14,⫺15,
⫺15,⫺16,⫺17,⫺19,⫺20,⫺21,⫺22 [cp6]
919 1y3m 4 4 100(4n),XXYY,der(1)t(1;17)(p36;q21)⫻2,⫹5,⫹9,⫹12,⫹13,⫹16,⫹18,
⫹18,⫹18,dmin
1031 1y9m 2 4 88–89(4n),add(1)(p13)⫻2,inc [cp4]
1185 5y7m 4 3 44,X,⫺Y,der(1)t(1;7)(p32;q11.2),der(17)t(1;17)(q21;p11),add(19)(p13),
⫺20/89(4n),XX,⫺Y,⫺Y,der(1)t(1;7)⫻2,der(17)t(1;17)⫻2,add(19)⫻2,⫺20
1187 1y9m 4 4 88(4n),XYY,⫺X, ?der(1)t(1;17)(p32;q12)⫻2,⫹add(5)(p14),⫺8,⫺8,⫺14,
⫺16,⫺22,⫹mar,dmin
1193 2y2m 4 4 83–87(4n),XY,⫺X,⫺Y,add(1)(p32),⫺2,⫺11,⫺22 [cp14]
*The study published by Kaneko et al., 1999 is partly based on the data presented in this table.
a
The karyotypes, which have never been published, are described on the basis of 92 (4n) chromosomes, because tetraploidization of diploid cell with
one 1p- chromosome and subsequent chromosome loss are easily recognized.

TABLE 3. Clinical, Cytogenetic, and Genetic Characteristics of Mass Screening (MS)-Positive and Mass
Screening-Negative/Late-Presenting Neuroblastomas Classified by the Constitution of Chromosome 1*
MYCNd
Number Siteb Stage of disease LOH on 1pc amplification 5-year EFSe
Group of of
patientsa patients Adr. Non-adr. 1 2 4S 3 4 ⫺ ID TD ND ⫹ ⫺ (%) 95% CI

Patients with MS-positive tumor

Dis1Norm1p 11 6 5 2 4 2 1 2 8 1 1 1 0 11 90.9 73.9–107.9
Dis1Del1p 8 7 1 2 2 2 0 2 1 0 3 4 2 6 50.0 5.3–94.7
Tris1Norm1p 151 82 69 64 43 11 26 7 65 6 2 78 0 151 98.7 96.9–100.5
Tris1Del1p 16 12 4 8 2 3 2 1 8 0 1 7 0 16 100.0 100.0–100.0
Total 186 107 79 76 51 18 29 12 82 7 7 90 2 184 96.3 93.3–99.3
Patients with MS-negative/late-presenting tumor
Dis1Norm1p 11 4 7 0 0 0 2 9 5 0 0 6 0 11 18.2 ⫺4.6–41.0
Dis1Del1p 33 29 4 0 0 0 5 28 5 0 13 15 22 11 12.1 ⫺1.0–25.2
Tris1Norm1p 13 5 8 2 2 0 4 5 9 1 0 3 3 10 69.2 44.1–94.3
Tris1Del1p 3 3 0 0 0 0 1 2 2 1 0 0 0 3 66.7 13.3–120.0
Total 60 41 19 2 2 0 12 44 21 2 13 24 25 35 28.3 16.3–40.4

*Kaneko et al., 1999.

a
Patients were grouped on the basis of the constitution of chromosome 1 in their tumor cells (see the text).
b
Adr., adrenal; Non-adr., non-adrenal.
c
⫺, no LOH on 1p; ID, interstitial deletion; TD, terminal deletion; ND, not determinable or not done.
d
MYCN amplification⫹, MYCN ⬎ 1 copy; MYCN amplification ⫺, MYCN 1 copy.
e
EFS, event-free survival; CI, confidence interval.

somy 1 with no 1p deletion), a Dis1Del1p group
(disomy 1 or tetrasomy 1 with 1p deletion), a
Tris1Norm1p group (trisomy 1, pentasomy 1, or a
mixed population of cells with trisomy and tetra-
somy 1, all without 1p deletion), and a Tris1Del1p
group (the same copy number of chromosome 1 as
the Tris1Norm1p group but with 1p deletion) (Ta-
ble 3) (Kaneko et al., 1999). Event-free survival
was lowest in the Dis1Del1p group, highest in the
Tris1Norm1p and Tris1Del1p groups, and inter-
mediate in the Dis1Norm1p group.
Of the 246 tumors, 186 were found by MS, and
60 were negative at MS but later found clinically.
Although 90% of MS-positive tumors were in the
two Tris1 groups, 73% of MS-negative/late-pre-
senting tumors were in the two Dis1 groups. Thus,
92 KANEKO AND KNUDSON
MS-positive tumors, many of which would have
regressed spontaneously, are characterized by an
unexpectedly high incidence of trisomy 1, and by
near-triploidy.
Relationship Between Ploidy and Other Genetic
Abnormalities
MYCN copy numbers were examined in the 246
tumors in the study described above (Kaneko et al.,
1999). MYCN amplification, a bad prognostic sign,
was frequent in the Dis1Del1p (58%) group, but
rare in the Tris1Norm1p (2%), Tris1Del1p (0), or
Dis1Norm1p (0) groups. Cytogenetic 1p deletion,
also a bad prognostic factor, was far more frequent
in Dis1 tumors (65%) than in Tris1 tumors (10%).
Terminal allelic losses (LOH) on 1p were found in
one of the 15 (7%) Dis1Norm1p tumors, 16 (73%)
of the 22 Dis1Del1p group, two (2%) of the 83
Tris1Norm1p tumors, and one (8%) of the 12
Tris1Del1p tumors. Thus, heterozygosity on ter-
minal 1p was retained in 81 (98%) of the 83
Tris1Norm1p tumors and 11 (92%) of the 12
Tris1Del1p tumors.
The high percentage of heterozygosity retained
in Tris1 tumors may be explained by a need to
delete two, rather than one, copies of 1p loci in
Tris1 tumors to produce allelic loss. Yet another
genetic factor affecting the biologic behavior of
neuroblastoma is gain of 17q. FISH and compara-
tive genomic hybridization (CGH) analyses have
shown that gain of 17q is detected in more than
90% of high-risk neuroblastomas, and cytogenetic
analysis has revealed that 17q participates in un-
balanced translocations with various chromosomal
partners, most often chromosome arm 1p, all lead-
ing to gain of the 17q23.1– qter (25 Mb) region
(Van Roy et al., 1994; Vandesompele et al., 1998).
17q gain simultaneously results in partial deletion
of the partner chromosome (Meddeb et al., 1996).
CGH analysis also showed an association of 17q
gain with near-diploid and near-tetraploid karyo-
types, and as expected an association of trisomy for
the entire chromosome 17 with a near-triploid
karyotype (Van Roy et al., 1994; Vandesompele et
al., 1998). Event-free survival is shorter in patients
with additional 17q than in patients without addi-
tional 17q (Caron et al., 1996; Bown et al., 1999). In
multivariate analysis, gain of 17q has been identi-
fied as the most powerful prognostic factor, fol-
lowed by the presence of Stage 4 disease and de-
letion of 1p (Bown et al., 1999). The aggressive
behavior of the tumors with an additional 17q sug-
gests that overexpression of genes on 17q, such as
NM23 or API4 (Leone et al., 1993; Adida et al.,
1998), and simultaneous deletion of putative tumor
suppressor genes located on the lost translocation
partner may give the tumor cells a proliferative
advantage.
CGH analysis on 23 neuroblastomas at Stages 3
and 4 also revealed frequent occurrence of 1p, 3p,
4p, 9p, 11q, and 14q deletions in addition to 17q
gain, and FISH analysis on the same tumors
showed disomy or tetrasomy 1 in the great majority
of the tumors (19/23) (Vandesompele et al., 1998).
Thus, chromosome arm deletions other than 1p are
also associated with diploid or tetraploid tumors.
Hypothesis Concerning Triploidy
How can one explain the drastic difference be-
tween the near-triploid, good-prognosis tumors and
the near-diploid, bad-prognosis tumors? Are the
genetic events totally different? Family studies
strongly suggest that the two groups of cases are
related, because different patients in affected fam-
ilies can have either form (Kushner et al., 1987).
CGH analysis on 6 familial neuroblastomas includ-
ing both early and advanced stage tumors showed
deletions of 1p, 3p, 10p, 10q, 11q, 16q, and 20q;
most of these sites are deleted also in sporadic
neuroblastomas (Van Roy et al., 1994; Altura et al.,
1997; Vandesompele et al., 1998). These data
would indicate that the initiating tumorigenic
event, at least in hereditary cases, and perhaps also
the secondary genetic events, are the same. One
possibility is that the first event in all cases is a
mutation at some chromosomal site, nearly always
occult. A mutation in, or loss of, the homologue
would then render the cell homozygously defective
at some critical locus, a “neuroblastoma gene,”
which would be a member of the recessive, tumor
suppressor, class of genes. Such a cell would be
diploid, and probably benign, but subject to other
genetic changes, notably mutation or loss of a gene
on chromosome arm 1p. Chromosome arm 1p has
been ruled out as a major mutational site for he-
reditary neuroblastoma (Maris et al., 1996). The
cytogenetic 1p deletion was a poor prognostic fac-
tor for diploid tumors, but not for triploid tumors
(Kaneko et al., 1999), suggesting that the prognos-
tic role of 1p deletion or the putative 1p36 gene
mutation may be determined by subsequent so-
matic genetic events.
As we described earlier, many neuroblastomas
have higher than normal ploidy states, sometimes
in the pentaploid range. We believe that these arise
from tetraploid cells, which in turn can arise di-
rectly from diploid cells by endoreduplication or
endomitosis. Tetraploid cells can undergo bipolar,
 PLOIDY AND NEUROBLASTOMA 93

Figure 1. Tripolar division of one tetraploid cell would have three results. (1) One diploid cell with
homozygous mutation (A), and two triploid cells, each with one mutant and two normal alleles; (2) one
diploid cell heterozygous for mutation, one triploid cell with one mutant allele, and one triploid cell with
two mutant alleles (B); (3) one diploid cell with no mutant alleles, two triploid cells with two mutant alleles
(B). Cell B will produce a tumor of Stage 1, 2, or 4S, and cell A will produce a tumor of Stage 4.

tetrapolar, or tripolar divisions in the following mi-
totic cycles. From bipolar divisions, the products
are two near-tetraploid cells. Tetrapolar divisions
give rise to four near-diploid cells, whereas tripolar
divisions produce two near-triploid and one near-
diploid daughter cells (Palitti and Rizzoni, 1972). It
has been noted that in the mitoses of hyperploid
cells, spindle formation may be defective, in which
case segregation will be incomplete. Thus, such a
defect in a tetraploid cell undergoing a tripolar
division could lead to the production of one near-
triploid and one near-pentaploid cell. We classified
triploidy that arose through tetraploidization and
succeeding tripolar division as “de novo,” and trip-
loidy derived from tetraploidization and chromo-
some loss as “secondary.”
Misdivisions of tetraploid cells have relevance for
the genetics of neuroblastoma. If a diploid cell is
heterozygous for mutation in a tumor suppressor
gene, then the tetraploid cell derived from it will
carry two mutant and two normal copies of the critical
chromosome, and, after DNA synthesis in the next
cell division, four mutant and four normal chromo-
somes. Bipolar division would produce two tetraploid
cells, each with two mutant and two normal chromo-
somes. Tetrapolar division can obviously have two
different outcomes: (1) four diploid cells, each het-
erozygous for the mutant chromosome, or (2) four
diploid cells, two heterozygous, one homozygous for
the normal allele, and one homozygous for the mu-
tant allele. The last cell would give rise to diploid
tumor cells, with no normal allele. Tripolar division
would have three different results (Fig. 1): (1) one
diploid cell homozygous for mutation, and two trip-
loid cells, each with one mutant and two normal
alleles; (2) one diploid cell heterozygous for the mu-
tation, and one triploid cell with one mutant, and one
triploid cell with two mutants; and (3) one diploid cell
with no mutant alleles, and two triploid cells contain-
ing one normal and two mutant alleles. These cell
divisions should occur in the relative frequencies of
1:2:1, so that four such divisions would produce on
average 12 cells with constitutions as follows: 2n ⫹/⫹
(1), 2n ⫹/⫺ (2), 2n ⫺/⫺ (1), 3n ⫹/⫹/⫺ (4), and 3n
⫹/⫺/⫺ (4). In those cases where defective spindle
formation leads to a pentaploid (n ⫽ 115) cell, the
latter would contain either two or three copies of the
normal allele.
A question then arises concerning the phenotype
of these triploid cells; can there be a dosage effect?
Because heterozygous diploid cells are probably
not abnormal, the triploid cells with one mutant
chromosome would probably be normal. We pro-
pose, however, that the cell with two mutant and
one wild-type chromosomes will produce a tumor
of Stage 1, 2, or 4S, and that this difference be-
tween diploid and triploid cells is crucial to an
understanding of the differences among the stages
of this disease. The one normal allele in the trip-
loid cell exerts some suppressor effect that pro-
duces a tumor of low malignancy. The assumption
is supported by the data presented in Table 3; viz.,
135 (74%) of 183 Tris1 tumors are at Stage 1, 2, or
4S, and 41 (65%) of 63 Dis1 tumors are at Stage 4.
Of 15 patients with Stage 4 Tris1 tumors, 10 were
alive with no evidence of the disease, and the other
5 died of the disease or of therapy-related compli-
cation. The former 10 tumors with favorable prog-
nosis were in keeping with their triploid state, and
the 5 tumors with poor prognosis may have lost the
one wild-type suppressing allele by non-disjunc-
tion, deletion, recombination, or mutation, predis-
94 KANEKO AND KNUDSON
posing to progression to Stage 4. Thus, most Dis1
tumors begin as Stage 4, and a small portion of
Tris1 tumors progress from early stages to Stage 4.
If the function of the wild-type “neuroblastoma
gene” were to suppress a neuroblast growth-pro-
moting gene, on another chromosomal region, then
heterozygous mutant diploid cells would have one
normal suppressor of two copies of this oncogene
(1:2) and a normal phenotype, whereas homozy-
gously mutant diploid cells would have none (0:2),
and be highly proliferative. Triploid cells would
have either two or one suppressors of three copies
of this other gene (2:3 or 1:3), and one could hy-
pothesize that the latter (1:3) could be abnormal
because of insufficient regulation of the three cop-
ies of the growth gene by a single copy of the
suppressor gene. Such a cell would be less prolif-
erative, however, than the diploid cell with two
mutated copies. Similarly, the pentaploid tumor
with just two normal alleles to suppress five copies
of some other gene (2:5) might be abnormal, but of
low growth potential.
Loss of a gene on chromosome arm 1p, which is
often accompanied by multiplication of a gene on
chromosome arm 17q, seems to be associated with
MYCN amplification, and necessary for high-grade
malignancy. Diploid tumors are far more likely
than triploid tumors to show such loss or gain. Two
factors are operating: (1) greater growth potential of
diploid cells, and (2) the need to mutate or lose just
two, rather than three, copies of this gene. Such
diploid tumors with one 1p- chromosome often
evolve to tetraploid tumors with two of the same
1p- chromosomes, probably through endoredupli-
cation or endomitosis, as described in the previous
section and elsewhere (Kaneko et al., 1985; Shack-
ney et al., 1989).
Predictions that follow from this hypothesis are
that linkage studies in familial cases should show
linkage with the same syntenic markers regardless
of stage, that early-stage triploid tumors should
contain one, and only one, normal copy of some
critical suppressor gene, and that late-stage diploid
tumors should show no normal copies of that same
gene. The identity of this gene should be revealed
by the study of hereditary cases: indeed, a recent
report localized a linked gene to chromosome band
16p12 (Maris et al., 1999). The only cases that
might benefit from MS are the diploid or tetraploid
tumors before dissemination, and the triploid and
pentaploid tumors before they lose a third copy of
the “neuroblastoma gene.” Triploidy occurs pri-
marily in tumors appearing in the first year of life,
apparently because triploid tumors without 1p loss
or MYCN amplification usually regress spontane-
ously beyond this age (Ambros et al., 1996). The
“neuroblastoma gene” may negatively regulate a
growth-promoting gene, whereas the gene at chro-
mosome arm 1p suppresses malignant progression.
Our hypothesis may be applicable to tumors
other than neuroblastoma, and testicular germ cell
tumor, which usually shows near-triploidy (de Jong
et al., 1990) and has cure rates exceeding 80%
(International Germ Cell Cancer Collaborative
Group, 1997), would be one of the candidates.
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