[CANCER RESEARCH 41. 4678-4686. November 1981]0008-5472/81 70041-OOOOS02.00

Cytogenetic Features of Human Neuroblastomas and Cell Lines1

Garrett M. Brodeur,2 Alexander A. Green, F. Ann Hayes, Kenna J. Williams, Dorothy L. Williams, and

Anastasios A. TsiatisDivisions of Hematology-Oncology ¡G.M. B.. A. A. G.. F. A. H.], Pathology [K. J. W., D. L. W.J, and Biostatistics ¡A.A. T.], St. Jude Children's Research Hospital,

Memphis, Tennessee 38101

ABSTRACT

We reviewed the banded karyotypes of 24 human neuro-

blastomas and cell lines to identify any consistent chromosomalabnormalities. Six of the 10 primary tumors and one of the 14cell lines were studied at this institution. Of the 24 neuroblas-tomas karyotyped, 20 were near-diploid, one was near-triploid,and 3 were near-tetraploid. One primary tumor had a diploid

karyotype without numerical or structural rearrangements. The20 cases with a karyotype in the diploid range were statisticallyanalyzed for gain or loss of whole chromosomes and forstructural abnormalities of each chromosome arm. The shortarm of chromosome 1 was preferentially involved in structuralrearrangements, occurring in 14 cases (p < 0.01). In 11 ofthese cases, the abnormality of chromosome 1 included deletion of bands 1p32 —¿�»1pter, rendering the cells monosomic

for this genetic material. Of the remaining three cases, oneinvolved a reciprocal translocation of chromosomes 1p and12q, another had insertion of genetic material at band 1p13,and the third had an extra dark band at 1p36. No othernumerical or structural abnormalities occurred with sufficientfrequency to reach statistical significance (p > 0.20). Six ofthe primary tumors or cell lines in the diploid range had doubleminute chromatin bodies, four cell lines had homogeneouslystaining regions, and two cell lines had either double minutechromatin bodies or homogeneously staining regions in sub-

populations of cells. Hence, partial monosomy for the shortarm of chromosome 1 was the most consistent cytogeneticabnormality in the human neuroblastomas studied.

INTRODUCTION

Cytogenetic analysis of the banded karotypes of humancancer cells has revealed specific, subtle abnormalities ofchromosome structure that were not apparent in routine preparations. Consistent chromosomal abnormalities have beenidentified in certain malignant diseases, especially the leuke-

mias (29). Solid tumors have been less extensively studiedbecause of technical difficulties in obtaining and processingtumor tissue. Nevertheless, as more tumors are analyzed withbanding techniques, disease-specific chromosomal abnormal

ities are becoming more apparent (29, 37).Three major cytogenetic abnormalities have been noted in

human neuroblastomas. (a) DM3 have been found in some

1This work was supported in part by American Cancer Society Grant IN-99E,

Clinical Cancer Education Grant CA-23944, Solid Tumor Program Project GrantCA-23099, National Cancer Institute BiomédicalResearch Support Grant RR-05584, Rubi-Levi Research Fund, and ALSAC.

2 Present address: St. Louis Children's Hospital, 500 South Kingshighway

Boulevard. P. O. Box 14871, St. Louis, Mo. 63178. To whom requests torreprints should be addressed.

3 The abbreviations used are: DM. double minute chromatin bodies; HSR,

homogeneously staining regions.Received September 3, 1980: accepted August 10, 1981.

neuroblastomas (3, 4, 9-12, 39, 41). However, they can also

occur in a variety of other tumors (4, 33, 37). (b) Giant markerchromosomes with HSR have been described for cell lines ofneuroblastoma (3, 4, 9-12, 41). HSR have also been reported

in cell lines of colon carcinoma (33) and breast carcinoma (7)as well as other cancers of breast, eosphagus, and pharynx(27); they usually occur in cell lines and do not involve anyhuman chromosome consistently, (c) A structural abnormalityof the short arm of chromosome 1 has been described in 2primary neuroblastomas and 1 of 4 neuroblastoma cell lines(12), but rearrangement of chromosome 1 has been reportedin other malignant diseases (1, 2, 14, 19, 26, 36-38, 43, 46).

Moreover, the prevalence of these and other chromosomalaberrations has not been established from a large number ofcases.

Since the first observations on banded karyotype abnormalities in neuroblastomas were published (3,10,12), the numberof such reports has almost tripled. Therefore, the purpose ofthis study was to review the banded karyotypes of 24 humanneuroblastomas and neuroblastoma cell lines for which datawere available. We determined the frequency of DM and HSRin these tumors and cell lines and statistically analyzed thekaryotypes in the diploid range for any structural or numericalrearrangements that were preferentially increased. Specifically, we sought additional evidence for involvement of theshort arm of chromosome 1 and determined the specific typesand sites of chromosome 1p abnormalities.

MATERIALS AND METHODS

Material for chromosome analysis at this institution was obtainedeither at the time of diagnostic surgery or when bone marrow wasaspirated from children with disseminated neuroblastoma. Cell suspensions were prepared from tumor tissue by mechanical dissociation andcultured in plastic Falcon flasks with liquid media [Roswell Park Memorial Institute Medium 1640, 10% fetal calf serum (KC Biologicals,Inc.), penicillin (100 units/ml), and streptomycin (100 units/ml)]. Suspension cultures were placed in a humidified atmosphere of 5% CO2and 95% air and harvested for chromosome analysis after 1 to 3 daysby a modification of the technique of Moorhead ef al. (31). Bandingwas done by a modified trypsin-Giemsa technique (40). Cytogenetic

analysis was attempted with tumor material from 24 different patients,of which 6 could be completely analyzed. The remaining 18 sampleswere inadequate because of low viability or mitotic index (W = 13) orbecause of poor definition of banded chromosomes (A/ = 5). In additionto the 6 primary tumors, 1 neuroblastoma cell line was completelyanalyzed. For each of these cases, at least 100 metaphases werecounted and 50 representative metaphases were photographed foranalysis.

To better determine the frequency of numerical and structural chromosome abnormalities for neuroblastomas, we combined our cytogenetic findings for 7 cases with those for 15 neuroblastomas reportedelsewhere (3, 4, 9, 10, 12, 16, 41) and 2 others which have not yet

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been reported in detail (13, 45):' Modal karyotypes from the 20 cases

in the diploid range were assessed for gain or loss of whole chromosomes and for structural abnormalities of the short or long arm of eachchromosome. For this analysis, modes of 35 to 57 were considered tobe near-diploid.

Chromosome gain, loss, or structural rearrangement were analyzedby a computer-simulated model. For this analysis, we assumed that (a)

the expected frequency of chromosome gains and losses would berandom (i.e., all chromosomes would be involved with equal frequency),and (£>)structural rearrangements would involve chromosomes directlyproportional to their size. These assumptions are generalizations basedon the occurrence of these respective abnormalities in normal cells.5

A computer program (21) was used to calculate the expected distribution of abnormalities based on observed number of abnormalities ofeach type and the assumptions given above. The maximum expectedfrequencies of numerical and structural rearrangements were thencompared to the observed frequencies to identify significant (p < 0.05

or 0.01) nonrandom involvement.

RESULTS

Neuroblastoma Tissue. Six of 10 primary neuroblastomaswere analyzed at this institution.6 NGO was obtained from a

newborn with an abdominal neuroblastoma and liver involvement at the time of diagnostic surgery. The tumor was analyzedby direct preparation, and all cells had a normal diploid kary-

otype without apparent numerical or structural rearrangementsor DM (Fig. 1). NBB was obtained from the bone marrow of a2.5-year-old with 100% marrow replacement by metastatic

neuroblastoma. The modal karyotype had 46 chromosomeswith a small deletion of the long arm of chromosome 6 in mostcells (Fig. 2), and 2% of the cells contained DM. NMB2 wasobtained from the bone marrow of a 2-year-old with recurrenttumor. The modal karyotype had 45 chromosomes with loss ofa chromosome 21. In addition, there were structural abnormalities of the short arm of chromosome 1, the long arm ofchromosome 8, and the long arm of chromosome 14 (Fig. 3).NJF was obtained from bone marrow of a 4-year-old with

metastatic neuroblastoma. The modal karyotype had 45 chromosomes with gain of a chromosome 14, loss of chromosomes15 and 18, and structural abnormalities of chromosomes 1p,6q, and 11q (Fig. 4). NKP was obtained from the bone marrowof a 5-year-old. The modal karyotype had 53 chromosomes

with a variety of numerical and structural abnormalities, including an insertion in the short arm of chromosome 1 (Fig. 5).NWC was obtained from a 5-year-old and had a modal kary

otype of 82 chromosomes. There were 2 to 4 copies of eachchromosome. In addition, there were 2 copies of each of thefollowing deletions: 1p-, 10q-, and 17p- (Fig. 6).

The 6 neuroblastomas analyzed above were combined with4 cases reported elsewhere (12, 13, 16) to determine thefrequency of the 3 most commonly reported cytogenetic abnormalities associated with neuroblastomas: DM, HSR, andstructural abnormalities of 1p (Table 1). Of the 10 cases, 7 hada structural abnormality of 1p, 3 had DM, and none had HSR.

Table 1

Selected cytogenetic findings for 10 primary neuroblastomas

Primarytumor1.

NGO2.NBB3.NMB4.NJF5.NKP6.NTP(12)7.NCC(12)8.

MBS(13)"9.

NKO(16)10.NWCMode

Abnormal 1pDM4646

+45+45

++53+46+46+44+4682

+ +HSR-—-——-—-——

' J. M. Trent, personal communication.5 G. M. Brodeur, A. A. Tsiatis, D. L. Williams. F. W. Luthardt, and A. A. Green.

Statistical analysis of chromosome abnormalities in human cancer cells, manuscript in preparation.

6 Preliminary interpretation of the karyotypes of NGO, NBB, NMB2, NKP, and

NCG was presented elsewhere (11). The interpretations presented herein containmodifications based on subsequent review and the availability of additionalkaryotypic information.

—¿�,absent; +, present.

Neuroblastoma Cell Lines. One of the 14 cell lines wasanalyzed at this institution.6 NCG had a mode of 45 chromo

somes with loss of a chromosome 3. In addition, there wasdeletion of the short arm of chromosome 1, a translocationinvolving chromosomes 9p and 15q, another involving 1q and9q, as well as a heterochromatic homogeneously stainingregion involving 16p (Fig. 7). The frequency of 1p structuralabnormalities, DM, and HSR for the 14 cell lines (our case and13 previously reported cases) is shown in Table 2. Ten celllines had a structural abnormality of 1p, 5 had DM only, 6 hadHSR only, and 2 had either DM or HSR in 2 subpopulations ofcells.

Analysis of Rearrangements. The 20 cases with karyotypesin the diploid range were pooled to assess the frequency ofnumerical and structural rearrangements, since a separateanalysis of tumors and cell lines gave identical results. Chart 1(fop) is a histogram showing gain or loss of whole chromosomesin the 20 cases. Statistical analysis indicated that the numericalchanges did not involve any chromosome with significant frequency (p > 0.20). We analyzed the short and long arm ofeach chromosome for structural aberrations, including deletions, translocations, inversions, insertions, and HSR (Chart 1,bottom). Fourteen (70%) of the 20 near-diploid neuroblastomas

had rearrangements of the short arm of chromosome 1. Theprevalence of involvement of either arm of other chromosomesby structural abnormalities was always <4 of 20 cases (<20%).

When analyzed according to the proportion of DNA in eachchromosome arm, the frequency of abnormalities of chromosome 1p was significantly increased (p < 0.01 ) but structuralabnormalities among the other chromosome arms did not reachstatistical significance (p > 0.20).

Chromosome 1p Abnormalities. We examined the specifictypes and sites of the abnormalities of chromosome 1p in the14 near-diploid cases as well as near-tetraploid tumor with 2

copies of the same 1p abnormality (NWC). In 12 of these 15cases, the distal portion of the short arm of chromosome 1 wasdeleted. Chart 2 is a schematic representation of chromosome1; brackets indicate the portion of genetic material identifiedas deleted in the 12 cases. Breakpoints occurred at 1p32 in 4cases, 1p31 in 4 cases, at 1p22 in 2 cases, and at 1p21 and1p13 in one case each. Although the breakpoint varied, theportion of chromosome 1 from p32 to pter appeared to bedeleted in all 12 neuroblastomas, and review of karyotypesfailed to identify this deleted material as translocated to anothersite. Of the remaining 3 cases with abnormalities of chromosome 1p, one (NAP) had a reciprocal translocation involvingthe distal portion of chromosome 1p and chromosome 12q,another (NGP) had translocation of material to 1p36, and the

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Table 2

Selected cytogenetic findings for 14 neuroblastoma cell lines

1.2.3.4.5.6.7.8.9.10.11.12.13.14.aCelllineNCGNGPÕ12)SK-N-SHO.

10)SK-N-MCO.10)SK-N-BE

(9, 10)IMR-32(9, 10)CHP-1

26(3)CHP-134(4)MCN-14NAP

(10)SMS-KANdO)NMD

(KOLA-N-1(9,41)LA-N-2O,41)-,

absent; +. present.Mode

Abnormal 1pDM45+"47

+4746

+44++49

+46+46

+-44

++45++46-f+8987

+73+ +HSR++——++++-——++-

WHOLE CHROMOSOME CHANGES

GAIN

9 IO II I2 lìI4 IS I6 I7 IB I9 2O 2I 22 X Y |

LOSS

STRUCTURAL REARRANGEMENTS

1

SHORT

9 IO II IZ I3 I4 I5 I6 I7 IS I9 2O 2I 22 X Y I

LONG

Chart 1. Whole chromosome gains or losses (top) and structural rearrangements of chromosome arms (bottom) in 20 human neuroblastoma tumors andcell lines with karyotypes in the diploid range. Each block represents a case withthe indicated chromosome abnormality. All gains and losses of whole chromosomes were randomly distributed ( p > 0.20). Structural rearrangements involvingthe short arm of chromosome 1 were significantly increased (p < 0.01 ). but otherchromosome arms were not preferentially involved (p > 0.20).

last (NKP) had insertion of genetic material at 1p13.DM and HSR. We analyzed the frequency of DM and HSR in

the near-diploid neuroblastomas and neuroblastoma cell lines.

Six tumors and cell lines had DM, and 4 other cell lines hadHSR. An additional cell line (CHP-126) had 2 subpopulations

of cells, one with DM and the other with homogeneouslystaining region (3, 4). In another case, 2 cell lines were established from the same patient at different times; one had DM

Chart 2. Human neuroblastoma chromosome 1 deletions. Schematic representation of chromosome 1 and brackets showing the portions of chromosome1p deleted in 12 human neuroblastoma tumors and cell lines. Breakpointsoccurred at 1p32 in 4 cases, 1p31 in 4 cases. 1p22 in 2 cases, and 1p21 and1p13 in 1 case each. The portion from p32 —¿�»pter appeared to be deleted in allcases, but the long arm of chromosome 1 was not affected.

and the other had HSR (9, 10). HSR have been reported toinvolve 18 of the 24 different chromosomes in human neuroblastomas, and no individual chromosome or chromosome armappears to be preferentially involved (9). Moreover, either DMor HSR (or both) were found in 16 of the 24 different neuroblastomas. DM were found in tumors and in cell lines, but HSRwere observed exclusively in cell lines.

DISCUSSION

Abnormality of the short arm of chromosome 1 was the mostcommon cytogenetic finding of human neuroblastomas basedon our analysis. Involvement of the short arm of chromosome1 occurred in 15 cases, including 12 that were monosomic forthe distal portion of chromosome 1p (p32 —¿�»pter). The fre

quency of numerical and structural abnormalities of other chromosomes or chromosome arms never reached statistical significance (p > 0.20). Either DM or HSR or both were found in16 of the 24 neuroblastomas and neuroblastoma cell lines. Ithas been postulated that these 2 entities are different manifestations of the same cytogenetic phenomenon (3-5, 9) and thatthey may represent gene amplification (8-10, 17, 42). Never

theless, abnormalities of 1p were about as common as DM andHSR combined and appear to be more specific for neuroblastoma, especially for primary tumor tissue.

Chromosome 1p abnormalities have been reported in othertumors of neutral crest origin. In meningioma, monosomy or

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Cytogenetic Features of Human Neuroblastomas

short arm deletion of chromosome 1 is second only to abnormality of chromosome 22 (37). In melanoma, the short arm ofchromosome 1 is frequently rearranged or missing (20, 28, 34,37), but the structural rearrangements are often complex, anda characteristic pattern of involvement has not yet emerged(37).

Rearrangements of chromosome 1 have been described fora variety of other tumors and cell lines. However, in most casesin which a 1p deletion was present, there were 2 normal copiesof chromosome 1, which indicates trisomy for the long arm ofchromosome 1. Trisomy 1q has been reported to be the mostcommon abnormality in myeloproliferative disorders (36) andin carcinoma of the breast (14, 26, 37), ovary (2, 37, 43),cervix (1, 37), and testis (46) and is reported in a variety ofother malignant diseases (19, 26, 37, 38). Indeed, this abnormality has also been noted in neuroblastomas [one primarytumor and 4 cell lines (9-12, 41)]. Trisomy 1q has beenobserved as a secondary change during the evolution of kary-

otype abnormalities in hématologieand ovarian cancers (2, 32)and has been correlated with resistant, progressive disease.Rowley (36) and Sandberg (37, 38) have suggested that trisomy 1q confers a selective advantage for proliferation, possibly by providing excess amounts of the gene products encoded in this portion of DMA. Its presence in such a variety ofmalignant diseases indicates that it is not specific for any onetissue or cell type but rather that it may be a common mechanism for enhancing malignant proliferation. In contrast, partial1p monosomy appears to be relatively specific for neuroblastoma.

The presence of a consistent chromosomal abnormality inneuroblastoma suggests that a causal relationship may existbetween the chromosomal abnormality and the neoplasm. It islikely that certain genes or gene products are responsible forthe regulation of growth and differentiation of neuroblasts. Ifsuch genes were deleted or rearranged, uncontrolled growthof these cells could result. Moreover, disease-specific chromosomal deletions or translocations have been found in avariety of human leukemias and solid tumors, suggesting thatthe gene(s) responsible for the regulation of different cell typesare located on specific chromosomes (29).

Chromosomal localization of tumor-specific genes has been

proposed for retinoblastoma and nephroblastoma (Wilms tumor) on the basis of constitutional chromosomal abnormalities.Retinoblastoma is associated with deletion of the long arm ofchromosome 13 (25, 47), and nephroblastoma is associatedwith deletion of the short arm of chromosome 11 (35, 48). Theconstitutional chromosome deletion syndromes in patients withthese tumors may be extreme examples of germinal mutationsthat predispose to specific cancers. While a distinguishingconstitutional deletion has not yet been identified in patientswith neuroblastoma (30), statistical analysis of incidence figures has shown that this tumor, like retinoblastoma and nephroblastoma, fits the 2-mutation hypothesis for the origin of

childhood cancer (22, 23). According to this hypothesis, thehereditary form arises in patients who inherit one mutation asa germinal (prezygotic) event, and all cells carrying this mutation would need only one additional mutation to induce malignant transformation. The nonhereditary form is the result of 2somatic (postzygotic) mutations in a single cell. Within thecontext of this model, a tumor-specific gene would result fromthe germinal or first somatic mutation. Evidence for the locali

zation of a retinoblastoma gene has been persuasive becausethe inherited cytogenetic abnormality appears to be the sameas the prevalent abnormality in tumor cells of patients withnormal karyotypes (6, 44). Although a hereditary deletion syndrome has not yet been demonstrated for neuroblastoma (15,30), our analysis of tumor cells suggests that a neuroblastomagene may be located in the short arm of chromosome 1.

By extension of his 2-mutation hypothesis, Knudson, with

Meadows (24), has proposed that some infants with neuroblastomas that regress spontaneously or respond well to lessintensive therapy may have only a single (germinal) mutation.This hypothesis predicts that these tumors with single mutations are hyperplastic nodules rather than clonal proliferationsof malignant cells. If this is true, then the karotype of suchtumors should be normal. Thus, the tumor with a normal kary-

otype (NGO, described in this report) may be an example of asingle-mutation tumor in an infant. The therapeutic approach

to these tumors perhaps should be aimed at the induction ofdifferentiation (24). Conversely, a patient of any age with aclone of cytogenetically abnormal neuroblasts clearly has acancer and probably should receive conventional therapy. Thecytogenetic analysis of neuroblastoma cells, therefore, mayprovide useful information to determine the appropriate therapyfor selected cases.

Since it is possible that a constitutional deletion syndromeexists for neuroblastoma, as identified for retinoblastoma andnephroblastoma, patients with neuroblastoma who are at highrisk for a germinal mutation should be studied. These individuals include: (a) patients with a family history of neuroblastoma;(b) those with multifocal primary tumors; (c) patients with dys-

morphic features or various expressions of a neurocristopathy;and (cOpatients less than 1 year old. These patients should bestudied by high-resolution banding of their blood lymphocytes,

with special attention paid to the short arm of chromosome 1.

ACKNOWLEDGMENTS

We are grateful to Drs. Alvin M Mauer and Alfred G. Knudson. Jr., for helpfulsuggestions in preparing this manuscript; to Drs. Jeffrey M Trent. June L.Biedler, Gloria Balaban-Malenbaum, C. Patrick Reynolds, and William F. Benedictfor sharing their banded neuroblastoma karyotypes for analysis; and to JaneSeifert for editorial review.

ADDENDUM

A recent article in Cancer Research (18) reports the results of serial karyotypeanalyses of a human neuroblastoma, examined after short- and long-term cultureas well as after passage in a nude mouse. Consistent cytogenetic abnormalitiesincluded 1p-, +17, -22, and DM. These results added to those reported here

do not change any of the conclusions but serve to strengthen the probablesignificance of partial 1p monosomy in neuroblastomas.

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Cytogenetic Features of Human Neuroblastomas

U u lì H

ii 31 U678

n n :: u9 10 11 12

AA13

:'« A«

14 15II16

U17 18

m19

II20 21

A«

22 X Y

Fig. 1. Karyotype of neuroblastoma designated NGO: 46.XY, without apparent numerical or structural abnormalities or DM

It13

ft19 20

8

li it14 15

9 10

**21 22

nIt If I! (I

11

fi as16 17

II

12

kl

18

I.X Y

Fig. 2. Karyotype of neuroblastoma NBB: 46,XY,del(6Xq25). (arrow). DM were present in 2% of cells.

NOVEMBER 1981 4683

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G. M. Brodeur et al.

IDI M II K123 45

» If IL ir u ?/6 7 8 9 10 11 12

M t¿_ II13 14 15 16 17 18

M |f •¿� ft* I«19 20 21 22 X Y

Fig. 3. Karyorype of neuroblastoma NMB2: 46.XY.-21, der(1;14X1qter-»1p32::14q31-»14qter),t(8; ?Xq24; ?),der(14) (pter—q24::q11—q24:). This representstranslocation between chromosomes 1 and 14 with deletion of 1p32—»1pterand direct duplication of 14q11—»14q24(arrows). The origin of extra material on the longarm of chromosome 8 is unknown.

ÕMr li O\ t

ÎÕ10 11

U n8 9

M iíl I i, « i13 14 15 16 17 18

«l H - II19 20 mär 21 22 XX

Fig. 4. Karyotype of neuroblastoma NJF: 45,XX, + 14,-15.-18,der(1Xqter-> p32::?p13-» p31:),del(6Xq23).del(11Xq23). This represents deletion of 1p32-»1pter with direct duplication of 1pt3—«1p31.Also shown are deletions of the long arm of chromosomes 6 and 11 (arrows). A single minute fragment of unknownorigin was seen in about one-half of the cells examined, and most cells contained numerous small but typical DM.

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Cytogenetic Features of Human Neuroblastomas

7*

n'-ilff H H f-

8 9 10 11 12

ti il13 14 16 17 18

19Ut20

È*21 22 XX

filmar 1 2 3

Fig. 5. Karyotype of neuroblastoma NKP: 53,XX, + 1,+4, + 15.+ 20, + mar1,+mar2. + mar3, der(1qter-»1p13::1p32-»1p13::1p13-»1pter),t(3;15Xq13.q26),del(4Xq31), t(7;1 1Xq36; q13). This represents duplication of bands 1p13-»1p32,deletion of the long arm of chromosome 4, translocations between chromosomes3 and 15 and between chromosomes 7 and 11 (arrows'), plus 3 unidentified marker chromosomes, and most cells contained numerous typical DM.

fi! ¡Ut'»li

6

5U ani8

aiir9

"-su10 11

«¿a12

13

19

14

Io*15

20 mar 1 2 21

16

22

17 18

««

illX X Y

Fig. 6. Karyotype of neuroblastoma NWC: 82,XXY.-3,-7,-1 1,-12.-15,-19.-19,-20,-21,-21,-22,-Y. + mar1,-l-mar2,del(1Xp22)x2,del(10Xq22)x2.deli 17Xp11)x2. This karyotype is expressed relative to a tetraploid male karyotype and shows numerous chromosome losses as well as 2 identical copies of deletionsof chromosomes 1p, 10q, and 17p (arrows), plus 2 small unidentified markers.

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M lìu-u «-{n

7 8 9 10 11 12

13 14 15 16 17 18

M ti M 14 1119 20 21 22 X Y

Fig. 7. Karyotype of neuroblastoma cell line NCG: 45,XY,-3.del(1Xp13).der(1q9qX1qter-.1q11::9p11-*9qter),der(9p15qX9pter-»9p11::15p11-»15qter),der(16p) (HSR) This represents deletion of 1p, translocation between 9p and 15q. translocation of extra 1q to 9q, and a heterochromatic homogeneously stainingregion involving 16p (arrows).

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1981;41:4678-4686. Cancer Res   Garrett M. Brodeur, Alexander A. Green, F. Ann Hayes, et al.   Cytogenetic Features of Human Neuroblastomas and Cell Lines

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