Download Chromosome Dosage Dependency1

[CANCER RESEARCH 44, 3471-3479,
August 1984]
Tumorigenicity of Human HT1080 Fibrosarcoma x Normal Fibroblast Hybrids:
Chromosome Dosage Dependency1
William F. Benedict,2 Bernard E. Weissman,3 Corey Mark, and Eric J. Stanbridge4
Clayton Molecular Biology Program and Division of Hematology-Oncology, Childrens Hospital of Los Angeles, Los Angeles, California 90027 [W. F. B., C. M.]; Department
of Microbiology, College of Medicine, University of California, Irvine, California 92717 [E. J. S., B. E. W.J: and Department of Pediatrics, University of Southern California
School of Medicine, Los Angeles, California 90027 [W. F. B.]
ABSTRACT
The tumorigenic capacity of hybrids formed by fusion of the
highly tumorigenic HT1080 human fibrosarcoma cell line with
nontumorigenic normal fibroblasts was examined. The HT1080
also contains an activated N-ras oncogene. Near-tetraploid hy
brids which contained an approximately complete chromosomal
complement from both parental cells were nontumorigenic when
1 x 107 cells were injected s.c. into athymic (nude) mice, whereas
the parental HT1080 cells produced tumors in 100% of the
animals with no latency period following injection of 2 x 106 cells.
Tumorigenic variants were obtained from these hybrids which
had lost only a few chromosomes compared to cells from the
nontumorigenic mass cultures. In addition, several near-hexaploid hybrids were obtained which contained approximately a
double chromosomal complement from the HT1080 parental line
and a single chromosomal complement from the normal fibro
blasts. All of these near-hexaploid hybrids produce tumors in
100% of nude mice with no latency period.
Our results indicate that tumorigenicity of these particular
human malignant cells of mesenchymal origin can be suppressed
when fused with normal diploid fibroblasts. In addition, the results
suggest that tumorigenicity in this system is chromosomal dos
age dependent, since a diploid chromosomal complement from
normal fibroblasts is capable of suppressing the tumorigenicity
of a near-diploid but not a near-tetraploid chromosomal comple
ment from the tumorigenic HT1080 parent. Finally, the loss of
chromosome 1 (the chromosome to which the N-ras oncogene
has been assigned) as well as chromosome 4 was correlated
with the reappearance of tumorigenicity in the rare variant pop
ulations from otherwise nontumorigenic near-tetraploid hybrid
cultures. Our results also suggest the possibility that tumori
genicity in these hybrids may be a gene dosage effect involving
the number of activated N-ras genes in the hybrids compared to
the gene(s) controlling the suppression of the activated N-ras
genes.
INTRODUCTION
The characterization of the genetic basis for tumorigenicity,
particularly in human cells, may be of fundamental importance in
understanding the oncogenic process in humans. One approach
to study the regulation of tumorigenicity is to examine various
'This
work was supported
by Grants CA19401
from the National Cancer
Institute and 1475R1 from the Council for Tobacco Research, Inc. The studies
were done in part in conjunction with the Clayton Foundation for Research.
2 To whom requests for reprints should be addressed, at the Division of
Hematology-Oncology, Childrens Hospital of Los Angeles, 4650 Sunset Blvd., Los
Angeles, CA 90027.
3 Present address: Division of Hematology-Oncology, Childrens Hospital of Los
Angeles, Los Angeles, CA 90027.
4 Recipient of Research Career Development Award KO4 CA00271.
Received August 8, 1983; accepted May 1,1984.
AUGUST
intraspecific human cell hybrids between tumor and normal cells
for their ability to produce tumors in immunosuppressed animals.
We have shown previously that the fusion of tumor cells of
epithelial origin with normal fibroblasts or keratinocytes results
in the total suppression of tumorigenicity (9, 15). The chromo
somal complements of both parental cells are generally main
tained, although the initial loss of a few chromosomes can occur
randomly (15).
Since the previous studies had utilized human tumor cells of
epithelial origin, namely HeLa cells, we wished to determine
whether a similar suppression of tumorigenicity occurs if a tumor
cell line of mesenchymal origin is used for fusion. The HT1080
fibrosarcoma cell line was chosen for this study, since it rapidly
produces fibrosarcomas in nude mice and has only a few chro
mosomal changes from the normal diploid karyotype (11). In
addition, the HT1080 cells contain an activated N-ras transform
ing oncogene which is located on chromosome 1 (5). In our initial
attempts at cell fusion, only true hybrids were obtained which
contained a near-double complement of HT1080 chromosomes
and a single complement of chromosomes from the normal
fibroblasts, resulting in a cell with a near-hexaploid chromosomal
number.5 We determined that all of the putative hybrids which
had a near-tetraploid chromosome complement were actually
pseudohybrids which had gained the ability to grow in selective
medium by way of gene transfer.5 Subsequently, after several
additional fusion attempts, 2 true hybrids were obtained which
had near-tetraploid chromosomal modes. The cytogenetic and
biochemical evidence that these cells represent true hybrids is
presented. The tumorigenicities of these near-tetraploid hybrids
are also reported and are compared to the tumorigenicities of
the near-hexaploid true hybrids. Evidence for the role of specific
chromosomes, particularly chromosome 1, in the suppression
and reexpression of tumorigenicity in this system also will be
presented.
MATERIALS AND METHODS
Cell Lines. The cell lines used for the various fusions are listed in
Table 1. Two diploid Lesch-Nyhan human fibroblast cell strains,
GM2291OR and 75-18OR, were used as the "normal" parental cells. These
cell strains are hypoxanthine-guanine
phosphoribosyltransferase
defi
cient and have been further selected for ouabain resistance. Clonal
derivatives of HT1080 cells were used for fusion with the fibroblast cells.
These included: HT1080-6TG C5, a hypoxanthine-guanine phosphoribosyltransferase-deficient variant cell line; HTD-114 CI, a pseudodiploid
adenine phosphoribosyltransferase-deficient
cell line (18); and HTD-114
MC4, a near-tetraploid adenine phosphoribosyltransferase-deficient
cell
line. All cell lines were grown ¡nEMEM6 supplemented with 10% fetal
calf serum (Flow Laboratories)
with glutamine and nonessential amino
5 B. E. Weissman, C. Mark, W. F. Benedict, and E. J. Stanbridge. Specific gene
transfer during whole cell fusion of human cells, submitted for publication.
6 The abbreviation used is: EMEM, Eagle's minimal essential medium.
1984
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3471
W. F. Benedict et al.
Table 1
Tumorigenicity and chromosomal modes of the parental cell lines
of mice with tu
mors/no, of mice
given injections0/6"
lineGM22910"
Cell
mode46
C2
75-1 8°"
46
HT1080-6TGC5
46
46°
HTD-114C1
HTD-114MC4Chromosomal
87No.
" One x 107 cells injected.
0 Two x 106 cells injected.
c Parental cell line contains a minor near-tetraploid
latency
(wk)0
0/6
6/66
6/6
6/6Av.
00
subpopulation which has 2
copies of each relevant marker chromosome.
acids added. All cell lines were tested frequently for the presence of
Mycoplasma contamination by both culture and 4',6-diamidino-6-phenylindole-2HCI assays (16) and were negative.
Cell Fusion. Somatic cell hybridization was performed by a modifica
tion of the method of Davidson and Gerald (4) as described previously
(19). The HT1080 derivatives and normal fibroblasts were plated in 60mm Petri dishes and incubated overnight. In general, 1 x 106 HT1080
cells were fused with 1 to 2 x 105 fibroblasts. Each dish was then treated
with a solution of serum-free EMEM containing 45% (M, 1000) polyeth
ylene glycol for 1 min. The dishes were then rinsed 3 times with serumfree EMEM, and growth medium was added to the cultures. Twentyfour hr after the fusion, the cells were trypsinized and replated into eight
100-mm dishes containing a selective medium consisting of growth
medium plus hypoxanthine-aminopterin-thymidine
and 5x10~7
M ouabain or 5 x 10~5 M ouabain alone. Dishes were refed every 4 to 5 days
with selective medium.
Selective Medium. The hypoxanthine-aminopterin-thymidine selec
tion of Littlefield (6) was coupled with 5 x 10~7 M ouabain to provide a
double selective medium (19). This medium was used for isolation of
hybrids from all fusions except for the HT1080-6TG x GM2291on C2
hybridization in which the selective was carried out only in the presence
of ouabain as described previously (19).
Chromosomal Analysis. Approximately 50 metaphase spreads/cell
line were photographed and counted to determine the chromosomal
mode. To evaluate the presence of specific marker chromosomes in
each of the parental cell lines and hybrids, the chromosomes were
banded using the Giemsa-trypsin banding technique as described previ
In contrast, 2 true hybrids, which were the result of the fusion
between 2 near-diploid or one near-tetraploid HT1080 cell and
one diploid fibroblast, were also obtained (SFTH-100 an SFTH101 ). They had modal chromosomal numbers of 127 and 131,
respectively, and contained 2 each of the HT1080 markers.
These 2 near-hexaploid hybrids produced tumors in 100% of
nude mice given injections (6 of 6 each) with no latency period.
In order to expand these initial observations, 2 different paren
tal cell lines were used for a second set of cell hybridization
experiments. HTD-114 CI is a pseudodiploid HT1080 subclone
which lacks adenine phosphoribosyltransferase activity. The hu
man fibroblast 75-18OR is a Lesch-Nyhan ouabain-resistant cell
line which synthesizes a different isoenzyme of glucose-6-phosphate dehydrogenase from the HT1080 cell line. True hybrid
cells can be identified in this fusion by both karyotypic analysis
and biochemical markers. The fusion of these 2 cell lines once
more produced only one near-tetraploid true hybrid (SFTH 400)
as judged by chromosomal content (Fig. 3A), presence of both
glucose-6-phosphate
dehydrogenase
isoenzymes, and the
expression of adenine phosphoribosyltransferase
activity. This
true hybrid also did not produce tumors following injection of 1
x 107 cells (Table 2), whereas the parental line HTD-114 CI
yielded tumors without any latency period in all animals after
injection of 2 x 106 cells (Table 1).
Subsequently after long-term culture, variant mass cell hybrid
populations were obtained (SFTH-300V and SFTH-400V) which
were tumorigenic, although there were minimal chromosomal
deviations from the parental SFTH-300 and SFTH-400 cell lines
(Table 2). These mass culture variants, nevertheless, produced
tumors with much longer latency periods than their HT1080
Table 2
Tumorigenicity and chromosomal modes of the near-tetraploid true hybrids and
variantsCell
their
of mice with
tumors/no, of mice
Latency
lineSFTH-3001SFTH-300VSFTH-300V-T1SFTH-300V-T2SFTH-300V-T3SFTH-300CL1SFTH-300CL2SFT
(wk)2001560=
somalmode8884756472828687808280898788878889898988898789898989No.
giveninjections0/64/56/6ND"ND0/12°0/1
ously (13).
Tumorigenicity Analysis. The tumorigenicity of the various parental
cell lines, the true hybrids, and the variant clonal derivatives of the true
hybrids was determined by s.c. injection of 2 x 106 to 1 x 107 cells in
nude (athymic) mice as described previously (17). The mice were exam
ined weekly for the appearance of s.c. tumors. No latency period indi
cates that palpable tumors were present from Day 1 which grew pro
gressively.
Glucose-6-phosphate
Dehydrogenase
Isoenzyme Analysis. Glucose-6-phosphate dehydrogenase analysis was performed as described
previously using cellulose acetate paper electrophoresis
2°0/1
2C0/1
2C3/12°0/12°0/55/116/6NDND0/1
(19).
RESULTS
Hybridization and Tumorigenicity Studies. Initial cell hybrid
ization studies were carried out using the HT1080-6TG C5 cell
line and GM2291OR C2 fibroblasts. The parental HT1080-6TG
C5 cell line is highly tumorigenic, producing tumors with no
latency period in all animals, whereas the diploid fibroblasts were
nontumorigenic. While many apparent hybrid cells were isolated,
only one cell line (SFTH-300) contained the correct modal chro
mosome number as well as the expected number of marker
chromosomes, indicating the fusion of one HT1080 cell with one
fibroblast hybrid cell (Fig. 2). This true hybrid produced no tumors
even when as many as 1 x 107 cells were inoculated (Table 2).
3472
2C0/1
2C0/12°0/1
2C0/12°0/1
2C0/60/60/60/6Av.
Parental HT1 080 lineChromo
HT1080-6TGC5.
3ND, not determined.
: Tumorigenicity tested independently in both of our laboratories (W. F. B., E. J. S.).
i Parental HT1080 line = HTD-114 CI.
CANCER
RESEARCH
VOL. 44
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Gene Dosage-dependent
parental cells. The SFTH-300V cell line yielded tumors in 4 of
the 5 mice given injections only after a latency period of approx
imately 20 weeks, and the SFTH-400V cell line produced tumors
in only 5 of 11 mice receiving injections after a latency period of
6 weeks.
These results suggested that there were minor revenant pop
ulations present in both near-tetraploid true hybrid cell cultures
which were tumorigenic. This was further suggested by the fact
that cells derived from tumors obtained from these variant cells
subsequently produced tumors with no latency period (Table 2).
In addition, we isolated 6 and 10 independently derived subclones from the SFTH-300 and SFTH-400 hybrids, respectively.
These were then tested for their tumorigenicity. No tumors were
produced by these subclones, except for one subclone (SFTH300-CL5) which gave tumors in 3 of 12 animals receiving injec
tions after a long latency period (Table 2), confirming the fact
that the SFTH-300V and SFTH-400V dérivâtes
represented only
minor tumorigenic revertant populations in an otherwise totally
suppressed near-tetraploid true hybrid culture.
Four additional independently derived near-hexaploid hybrids
were derived from fusion of a near-tetraploid HT1080 subclone
(HTD-114 MC4) with the 75-18OR diploid fibroblasts. These hy
brids (SFTH-200, -201, -202, and -203) had modal chromosomal
numbers of 113, 106, 108, and 111, respectively. They also
produced tumors in 100% of mice given injections (6 of 6 animals
for each hybrid) with no latency period. Such results suggested
that tumorigenicity in HT1080 x normal fibroblast hybrids is
chromosome dosage dependent, in that tumorigenicity was sup
pressed when equal chromosomal complements from each pa
rental cell line were present in the hybrids. However, tumori
genicity was retained if approximately twice the chromosomal
complement of the HT1080 derivative was present in the hybrid
cells compared to the chromosomal contribution of the normal
parent.
Chromosomal Studies. The karyotypes of the 2 pseudodiploid HT1080 derivatives used for the cell fusions are shown in
Fig.1. Both cell lines had retained the 2 HT1080 marker chro
mosomes reported previously (11), namely a No. 5 chromosome
with a small translocation onto the short arm and a No. 11
chromosome with a larger translocation onto the long arm. In
addition, the HT1080-6TG derivative contained a submetacentric
marker chromosome (P1), which represents a chromosome 7
with pericentric inversion, and the HTD-114 cell line had a
metacentric marker chromosome (92), which is an isochromosome for the long arm of chromosome 14. The near-tetraploid
true hybrid SFTH-300 contained one copy of both HT1080
Tumorigenicity
marker chromosomes as well as the P1 marker (Fig. 2; Table 3).
Four other marker chromosomes (HL H2, H3, and H4) which were
specific for this particular hybrid were also observed in all cells
(Fig. 2; Table 3). The variant mass cell culture obtained from the
SFTH-300 hybrid (SFTH-300V) retained all 3 marker chromo
somes present in the HT1080-6TG parental cell but contained
only one of the hybrid-specific markers (H!) which were present
in the SFTH-300 cell line. In addition, 2 new marker chromo
somes (Vi and V2) were present in all the cells examined (Table
3). These specific chromosomal changes within SFTH-300V
demonstrate a marked selection of a minor population which
was present in the SFTH-300 cell line. Furthermore, when the
chromosomal patterns of 3 independently derived tumors from
the SFTH-300V cells were analyzed again, each retained one of
the hybrid-specific markers (H,) and had, in addition, gained 4
common tumor-specific markers (Table 3). This latter finding
indicates that only one revertant cell population produced the 3
tumors from the variant SFTH-300V mass cell cultures injected.
A considerable number of chromosomes were lost in the
tumors compared to the variant cells injected (SFTH-300V) or
the nontumorigenic parental hybrid, SFTH-300 (Table 2). There
fore, the loss of several chromosomes could be correlated with
the reversion to tumorigenicity in these specific hybrids. Detailed
chromosomal analysis of the parental hybrid and the 3 tumors
derived from the mass cell culture variant demonstrated that the
loss of chromosomes 1, 2, 3q, 4, 9, 13, 17, 18, or Y or a
combination thereof could be correlated with the reversion to
tumorigenicity (data not shown).
The second near-tetraploid hybrid SFTH-400 and its tumori
genic revenants were considerably more useful for comparing
the loss of specific chromosomes with the reappearance of the
tumorigenic phenotype. The nontumorigenic SFTH-400 hybrid
contained one copy each of the HT1080 marker as well as the
HTD-114 parental marker, P2 (Fig. 3/4; Table 3). Again, a variant
mass culture population was obtained (SFTH-400V) which had
gained 3 variant-specific markers, V3, V4, and V5 (Fig. 3B; Table
3). All 3 tumors examined cytogenetically which were obtained
following injection of the SFTH-400V mass culture had a karyotype similar to that of the injected cells (Fig. 3C), indicating that
a rare event had occurred in the parental hybrid culture resulting
in the reappearance of tumorigenicity. Cytogenetic analysis of
the nontumorigenic SFTH-400 parental hybrid and 3 tumors
obtained from the SFTH-400V revertant population revealed only
a few chromosomal differences between the tumorigenic and
nontumorigenic hybrids (Table 4). Only a loss of chromosome 1
or chromosome 4 was found to be correlated with the reexpres-
TaWe3
Numberof specificmarkerchromosomesin tetrap/oidtruehybrids
Hybrid-specificmarkers
HT1080
Tfitraploiri hy
HJSFTH-300
brid cell lines
markers
B5
C11
P1 P2
H,
T41111111111111
markersV2
markersT,
T,
T3
11SFTH-300V
111
10SFTH-300V-T1
111
10SFTH-300V-T2
111
10SFTH-300V-T3
111
10SFTH-400SFTH-400VSFTH-400V-T1SFTH-400V-T2SFTH-400V-T3H310000H410000V,1000Variant-specific
111
None1
11
11
11
Vs1000Tumor-specific
V3
V4
1
11 1
11 1
11 1
11
1
1NoneNoneNone
AUGUST 1984
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3473
W. F. Benedict et al.
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3474
CANCER
RESEARCH
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VOL. 44
Gene Dosage-dependent
sion of tumorigenicity. It should be noted that the lost chromo
some 1 or chromosome 4 was among the several chromosomes
which correlated with the reversion to tumorigenicity in the SFTH
300 hybrid series as mentioned above.
Finally, each of the 6 tumorigenic near-hexaploid true hybrids
contained 2 copies of each HT1080 marker as well as 2 copies
of the specific parental markers (either P1 or P2), depending
upon which parental HT1080 derivative was used for the fusion.
This indicates that these near-hexaploid hybrids were formed by
the fusion of one near-tetraploid or 2 pseudodiploid HT1080 cells
with one normal fibroblast.
DISCUSSION
Previous studies done in one of our laboratories (E. J. S.) have
demonstrated that tumorigenicity is suppressed in hybrids re
sulting from the fusion of epithelial human tumor cells with normal
human fibroblasts or keratinocytes (9, 15). In addition, tumori
genic revenants appeared to have occurred as the result of
specific human chromosomal loss (17). The studies reported
here would also support the contention that suppression of
tumorigenicity occurs in hybrids formed between pseudodiploid
tumor cells of mesenchymal origin and normal diploid fibroblasts.
We have also found in this particular system that tumorigenic
ity (as measured by tumor formation in nude mice) appears to
be chromosome dose dependent. Six independently derived
hybrids which contained approximately twice the chromosomal
complement of the tumorigenic HT1080 parent were all highly
tumorigenic. In contrast, the cell hybrids which contained an
approximately equal chromosomal representation from both
HT1080 and normal human fibroblast cells were nontumorigenic.
Others (20) as well as ourselves (1, 2) have suggested several
years ago that tumorigenicity resulted from the balance of "expressor" and "suppressor" chromosomes. Our results utilizing
the HT1080 fibrosarcoma cell line would also suggest that there
may be a balance between chromosomes containing information
for the expression of tumorigenicity and chromosomes which
can suppress tumorigenicity. It is possible that there are insuffi
cient numbers of suppressor chromosomes in the tumorigenic
variant hybrids or in hybrids containing approximately twice the
number of chromosomes from the HT1080 parent compared to
the chromosomal complement from the normal human fibroblast.
It should be mentioned that the results of our studies are in
contrast to the previous report on the tumorigenicity of hybrids
formed between HT1080 fibrosarcoma cells and 2 separate
fibroblast lines, both of which had specific translocations (3).
Three near-hexaploid hybrids which apparently contained ap
proximately twice the chromosomal complement from the
HT1080 parent were tumorigenic (3). Although these findings
are similar to that reported by us, the authors also state that the
5 near-tetraploid hybrids they isolated were also highly tumori
genic (3). It is possible that each "near tetraploid" hybrid these
authors obtained produced tumors because they had lost one
or more suppressor chromosomes. No detailed karyotypic anal
ysis is presented in the paper (3). Consequently, it is not possible
to determine the number of No. 1 or No. 4 chromosomes present
in their "near tetraploid" hybrids.
The fact that a loss of chromosome
reexpression of tumorigenicity in our
of particular interest. The HT1080 cell
ing gene (7, 10) which has recently
AUGUST
1 was correlated with the
near-tetraploid hybrids is
line contains a transform
been named N-ras and
Tumorlgenicity
assigned to chromosome 1 (5). It is also the transforming gene
obtained from the SK-N-SH neuroblastoma line (7, 14) and from
the RD rhabdomyosarcoma cell line as well as from the promyelocytic leukemia HL60 cell line (7, 8). The possibility exists from
our studies that the normal No. 1 chromosomes in the fibroblasts
may contain a gene which could not only suppress tumorigenicity
in the system but also specifically regulate the expression of the
activated N-ras gene if the latter is correlated with the expression
of tumorigenicity. Such a gene may even be the normal N-ras
alÃ-eleand is consistent with speculation on the results from a
recent study where the normal alÃ-elewas lost in the tumor,
although in that study the activated oncogene was a K-ras rather
than a N-ras (12). Specific studies now in progress should allow
us to determine the role of chromosome 1 and, particularly, the
N-ras oncogene in the tumorigenic phenotype of the HT1080
fibrosarcoma.
ACKNOWLEDGMENTS
We wish to thank Joyce Wilkinson for excellent technical assistance and KeCheng Chen for his photographic assistance. We also thank Dr. Corsaro, Dr. Croce,
and Dr. Tischfield for gifts of parental cell lines.
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in human cell hybrids. Somatic Cell Genet., 7: 699-712, 1981.
18. Tishfield. J. A., and Stambrook, P. J. Interspecies and ¡ntraspecies transformation of the gene for adenine phosphoribosyl transferase. J. Cell Biol., 87:
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19 Weissman, B. E.. and Stanbridge, E. J. Characterization of ouabain resistant,
hypoxanthine phosphoribosyl transferase deficient human cells and their usefulness as a general method for the production of human cell hybrids. Cytogenet. Cell Genet., 28: 227-239, 1980.
20. Yamamoto, T., Rabinowitz, Z., and Sachs, L. Identification of the chromosomes
that control malignancy. Nat. New Biol., 243: 247-250,1973.
Fig. 1. Karyotypes of pseudodiploid HT1080 parental derivatives. Top, HT1080-6TGC5, a donai derivative of HT1080-6TG containing the 2 HT1080 marker
chromosomes (a No. 5 chromosome with an additional dark band on the short arm and a No.11 chromosome with a large translocation onto the long arm). An additional
marker chromosome (P,), which is a No. 7 chromosome with a pericentric inversion and the loss of a No. 7 chromosome, was observed in each metaphase analyzed
from the clone. Bottom, HTD-114 CI clonal derivative containing the same HT1080 marker chromosomes mentioned above as well as an additional marker (P2), which is
an iso 14q that was present in each metaphase. A No. 14 chromosome also was consistently missing.
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3477
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the parental HT1080-6TG marker chromosome (Pi). This indicates that the hybrids were formed by fusion of one HT-1080 and one normal fibroblast parental cell. Four
hybrid-specific marker chromosomes (H,, H2, H3, and H<) were present in all SFTH-300 metaphases. The origin of the Ht and H2 markers is not known. The H3 marker is
an ¡so1q chromosome, and the H, marker is a 9p+ chromosome.
Fig. 3. Modal karyotypes of near-tetraploid true hybrid SFTH-400 (A), its tumorigenic variant SFTH-400 (B). and a tumor produced by SFTH-400V, namely SFTH-400T1 (C). Note again that one copy of the 2 HT1080 marker chromosomes is present in all metaphases as is the HTD-114 C1 parental marker (P2).The variant tumorigenic
SFTH-400V cells also contained 3 new variant-specific markers (V3, Vt, and V5). The tumors derived from this variant had the same variant-specific markers present,
indicating the selection of a minor population of tumorigenic cells from the nontumorigenic SFTH-400 near-tetraploid hybrid. Marker V3 is a translocated chromosome in
which the long arm of chromosome 1 has translocated to the chromosome 15. The V»marker is an iso 1p chromosome, and the V5 marker is a Xp~ chromosome. The
2 unidentified chromosomes shown to the right of Marker P2 (A) are random chromosomal changes seen only in this specific metaphase.
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Tumorigenicity of Human HT1080 Fibrosarcoma × Normal
Fibroblast Hybrids: Chromosome Dosage Dependency
William F. Benedict, Bernard E. Weissman, Corey Mark, et al.
Cancer Res 1984;44:3471-3479.
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