Download Human Chromosome 16 Suppresses Metastasis

ICANCER RESEARCH 58. 4572-4576.
October 15. 1998]
Advances in Brief
Human Chromosome 16 Suppresses Metastasis But Not Tumorigenesis in Rat
Prostatic Tumor Cells1
Tomoyuki Mashimo, Misako Watabe, Andrew P. Cuthbert, Robert F. Newbold, Carrie W. Rinker-Schaeffer,
Eric Heifer, and Kounosuke Watabe2
Department iif Medical Microbiology and Immunology. Southern Illinois University. School of Medicine, Springfield. Illinois 62702 ¡T.M., M. W.. E. H.. K. W.I: Human Cancer
Genetics Unit. Department of Biology anil Biochemistry. Brunei University, Uxbridge. Middlesex UBK 3PH, United Kingdom IA. P. C. R. F. N.J: and Section of Urolog\.
Department of Surgen: University of Chicago. Chicago. Illinois 60637 ¡C.W. R-SJ
Abstract
<.i mimi, aberrations at the chromosome 16q arm are one of the most
consistent ahnormalities observed by loss of heterozygosity and compar
ative genomic hybridization analyses in human prostate cancer, suggest
ing that there are tumor suppressor or metastasis suppressor genes en
coded by this chromosomal
region. To functionally identify such
suppressor genes, we have conducted microcell-mediated
chromosome
transfer to introduce human chromosome 16 into the highly metastatic
Dunning rat prostatic cancer cell line, AT6.1. The metastatic ability of the
resultant microcell hybrid clones was then tested in a standard spontane
ous metastasis assay using SCID mice. When the microcell-mediated
chromosome transfer hybrid cells containing whole human chromosome
16 were injected, the number of metastatic lesions in the lung was signif
icantly reduced as much as 99% on average. Therefore, chromosome 16
has a strong activity to suppress the metastatic ability of AT6.1 cells while
it did not affect the tumorigenesis and tumor growth rate. A PCR analysis
of various microcell hybrid clones with sequence-tagged site markers
indicates that the metastasis suppressor activity is located in the q24.2
region of chromosome 16. Our results are consistent with the previous
finding that the region of human chromosome 16q has frequent loss of
heterozygosity in prostate cancer patients and suggest that there is a
metastasis suppressor gene in this region that may play an important role
in the progression of prostate cancer.
Introduction
Prostate cancer is the most common malignancy and second leading
cause of cancer death among men in the United States ( 1). However,
the molecular mechanisms underlying the prostate cancer tumorigenesis and tumor metastasis are still poorly understood. Chromosomal
abnormalities in prostate cancers on chromosomes 6. 7, 8. 10, 11, 13.
16. 17, and 18 have been observed frequently by LOH1 and CGH
analysis (2-8). These results suggest that there are tumor suppressor
or metastasis suppressor genes on these chromosomes that are in
volved in prostate carcinogenesis. In the past few years, rigorous
attempts have been made by several groups to identify the specific
genes involved in prostate tumor progression (9-11 ). A recent finding
of the familial prostate cancer susceptibility gene on human chromo
some 1(11) has provided a promising new avenue for prostate cancer
Received 3/4/98; accepted 8/28/98.
The costs of publication of this article were defrayed in pan by the payment of page
charges. This article must therefore be hereby marked advertisement in accordance with
18 U.S.C. Section 1734 solely to indicate this fact.
' This work was supported by Grant R15CA67290-01 from the NIH.
2 To whom requests for reprints should be addressed, at Department of Medical
Microbiology and Immunology. Southern Illinois University. School of Medicine. Spring
field. IL 62702. Phone: (217)782-3969;
Fax: (217)524-3227;
E-mail: kwatabe@
wpsmtp.siunied.edu.
1The abbreviations used are: LOH. loss of hctero/ygosity; CGH. comparative
genomic hybridi/ation: MMCT, microcell-mediated
combined immunodeficient; STS. sequence-tagged
bridi/ation.
research, although the identity and function of the gene remain to be
investigated.
We have chosen to pursue a functional approach to identify poten
tially important genes for cancer progression using the MMCT tech
nique. MMCT is particularly suitable for identifying tumor suppressor
and metastasis suppressor genes, and this powerful technique was
used recently to isolate a metastasis suppressor gene on human chro
mosome 11. When chromosome 11 was introduced into highly met
astatic rat prostate cancer cells by MMCT, the metastatic ability of the
resultant microcell hybrids was dramatically suppressed in athymic
nude mice while their tumor growth rate was unaffected (12). These
studies demonstrated that the 1Ipl3-pl 1.2 region of human chromo
some encodes novel metastasis suppressor genes. Subsequent posi
tional cloning and candidate gene study identified two genes, KAU
and CD44. in this chromosomal region as specific metastasis suppres
sor genes (12, 13). A similar approach is being used to identify
metastasis suppressor genes on human chromosomes 8, 10, and 17
(14-20), and these results have proven the utility of the functional
MMCT approach. Therefore, to take a more systematic approach, we
have recently constructed a highly stable human:rodent monochromosomal hybrid panel for an entire set of human chromosomes (21).
Each normal individual chromosome in the panel was tagged with a
selectable hygromycin marker so that they serve as donor sources for
a more systematic and functional screening of the tumor metastasis
suppressor genes.
Genetic imbalance of human chromosome 16q is most frequently
and consistently observed in sporadic prostatic carcinoma (2-8). The
results of a recent CGH analysis indicate that >55% of clinical
samples of metastatic prostate tumors have deletions in the q arm of
chromosome 16 (8). Therefore, it has been hypothesized that there are
tumor suppressor or metastasis suppressor genes on chromosome 16q.
To search for a potential metastasis suppressor gene, we conducted the
MMCT experiments using a newly constructed humanirodent monochromosome hybrid. Human chromosome 16 was transferred into a
highly metastatic rat prostatic carcinoma cell line, AT6.1, and result
ant microcell hybrids were tested for suppression of a spontaneous
metastatic ability to the lung in SCID mice. We found that the human
chromosome 16 has a strong capability of suppressing metastatic
ability of AT6.1 in vivo. A STS-based PCR analysis of the panel of
various microcell hybrid clones allowed us to locate the suppressor
activity in the q24.2 region of chromosome 16, which is distinct from
the location of the E- and P-cadherin genes. Our results suggest that
human chromosome 16q encodes a novel tumor metastasis suppressor
gene.
Materials and Methods
chromosome transfer; SCID. severe
site: FISH, fluorescence in situ hy
Cell Lines. AT6.1 is a highly metastatic. anaplastic. androgen-independent
rat prostatic cancer cell line that was established from a lung metastasis in the
Dunning R3327 rat model as described previously (14). The cell line was
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All seven clones (AT6.1-16-2, AT6.1-16-6. AT6.1-16-7, AT6.116-9, AT6.1-16-10, AT6.1-16-11, and AT6.1-16-12) containing chro
cultured and maintained in RPMI 1640 (Life Technologies, Inc., Gaithersburg.
MD) containing 10% PCS, 110 jig/ml streptomycin, 100 units/ml penicillin,
and 250 HMdexamethasone (PCS and antibiotics from Sigma Chemical Co., St.
Louis, MO). A9 (16) is an immortalized mouse A9 fibroblast cell line carrying
a single human chromosome 16 tagged with the selectable hygromycin resist
ance gene and was constructed as described previously (21). The donor cell
line was grown in DMEM supplemented with 10% PCS containing 400 ^.g/ml
hygromycin B (Life Technologies, Inc.) at 37°Cin 5% CO2 and 95% air.
mosome 16, as well as the parental cell AT6.1. were individually
injected s.c. into the dorsal flank of SCID mice. They were monitored
for tumor formation and their growth rates and were sacrificed at 4
weeks after inoculation of the cells. At the experimental end point,
their lungs were removed, and the number of macroscopic métastases
was examined. As summarized in Table 1, all of the mice injected
with the seven hybrid clones of chromosome 16 and the parental
AT6.1 cells formed tumors with similar growth rates during the
4-week period. Therefore, chromosomes 16 does not appear to have
any dominant effects on the tumorigenic ability of AT6.1 in mice. In
contrast, the metastatic ability of AT6.1 to the lungs is strongly
suppressed by human chromosome 16. As shown in Table 1, five of
the seven chromosome 16 hybrid clones, AT6.1-16-2, AT6.1-16-6,
AT6.I-16-7, AT6.1-16-10, and AT6.1-16-11, showed significantly
lower incidences of lung metastasis (40-70%) compared with the
parental AT6.1. The gross examination of the lungs of mice injected
with these five microcell hybrid clones of chromosome 16 revealed
that the number of metastatic lesions was also significantly reduced as
much as 99% on average (AT6.1-16-2). These results suggest that
human chromosome 16 has the ability to suppress the tumor meta
static phenotype of AT6.1 cells without affecting their tumorigenic
potential, implying that there is a tumor metastasis suppressor gene on
human chromosome 16.
To confirm the human origin of chromosome 16 in the hybrid
clone, we prepared metaphase cells from the AT6.1-16-2 clone and
MMCT. Microcell hybrids were generated as described previously (18).
Briefly, the monochromosomal
hybrid donor. A9 (16). was treated with 0.2
/ig/ml Colcemid followed by 2 /¿g/mlcytochalasin B. The resultant microcells were collected by centrifugation and then fused with the recipient
cell, AT6.I. using 50% polyethyleneglycol
(Sigma). Actively proliferating
individual clones were isolated and expanded in growth medium containing
hygromycin B.
STS-PCR Analysis of Hybrid Clones. The presence of human chromo
some 16 in hybrid clones and their spontaneous deletions were analyzed by
PCR using STS markers. High molecular weight DNAs were prepared from
A9 donor cells. AT6.1, and each hybrid clone, as well as tumor tissues from
metastatic lesions in the lungs of SCID mice, by proteinase K-SDS digestion
followed by phenol/chloroform extraction according to the standard proce
dures (22). All STS primers (MapPairs) were purchased from Research Ge
netics. Inc. (Huntsville, AL), and the E-cadherin exon 13 primers (5) and
P-cadherin exon 1 primers (23) were synthesized by Life Technologies, Inc.
(Gaithersburg. MD). Information regarding polymorphic loci for the markers
was obtained from the database of the Whitehead Institute (Massachusetts
Institute of Technology) and Genome Database. Isolated DNAs from the cells
were amplified with each set of primers by PCR with 35 cycles of denaturing
(94°Cfor 1 min), annealing (57°Cfor 1 min), and extension (72°Cfor 1 min).
PCR products were subjected to nondenaturing 8% PAGE and then stained
with ethidium bromide, followed by visualization under UV light.
Spontaneous Metastasis Assay. To characterize the in vivo growth rate
and metastatic ability of the microcell clones. 0.5 X IO6 cells in 0.2 ml of PBS
were s.c. injected in the dorsal flank of SCID mice. 5 weeks of age (HarÃ-an
Sprague Dawley. Indianapolis, IN). Mice were monitored daily, and tumor
volume was measured as an index of growth rate. Tumor volume was calcu
lated using the equation: Volume = (Width + Length)/2 X W X L x 0.5236.
Mice were sacrificed 4 weeks after the inoculation, and macroscopic métas
tases were counted visually.
FISH. FISH of metaphase cells from AT6.1-16-2 was performed as de
scribed previously (24) using the whole chromosome painting probe for human
chromosome 16 (WCP16 Spectrum Green; Vysis. Inc., Downers Grove, IL). A
total of 20 metaphases cells was scored for the presence of signals.
RT-PCR. From each clone of microcell hybrids, total RNA was isolated
using the RNeasy Total RNA system (Qiagen, Santa Ciarita, CA). Total RNA
from human skeletal muscle was purchased from Clontech (Palo Alto. CA).
All RNAs were treated with RNase-free DNase (Life Technologies, Inc.) and
repurified using the RNeasy Total RNA system. For RT-PCR, 5 ^g of RNA
were reverse-transcribed using random primers and a Moloney leukemia virus
reverse transcriptase (Perkin-Elmer, Foster City. CA). The cDNA was then
amplified with a pair of 5' and 3' primers for H- and M-cadherin genes
(25-27) in the reaction mixture containing deoxynucleotide
16
performed a FISH analysis using the whole chromosome painting
probe for chromosome 16. Fig. 1 shows a representative result of the
analysis. When 20 metaphase cells were examined, all of them re
tained a single copy of human chromosome 16. A counterstaining of
the chromosomes with 4',6-diamidino-2-phenylindole
indicated that
they were entirely composed of human sequences. More than 95% of
interphase cells also contained a single copy of human chromosome
16. These results suggest that the entire chromosome 16 was stably
maintained in AT6.1 cells and that a single copy of the chromosome
is enough to show the metastatic suppressor activity.
Structural Analysis of Chromosome 16 in Microcell Hybrid
Clones. To localize the metastasis suppressor activity on human
chromosome 16, high molecular weight DNAs were isolated from all
seven microcell hybrid clones of chromosome 16 (see Table 1), and
the lengths of the chromosomal fragments retained in these clones
were examined for various STS markers by PCR. As shown in Fig. 2,
A and B, the STS marker D16S402 was found to be commonly
retained in all five microcell hybrids that showed a significant reduc
tion in lung metastasis, whereas the same marker was lost from the
triphosphates and
Taq DNA polymerase. After the reaction, an aliquot of the product was
subjected to 8% PAGE. DNA products were visualized by staining them with
ethidium bromide and then photographed.
Table I In vivo characteristics
Tumorigenicity"ParentalAT6.1MMCT
Cell line
of parental and microcell hybrid clone*
métastases'"Mean
vim
doubling
SE'72±
time(days!3.02.72.73.33.62.82.52.3Lung
Results
Effects of Human Chromosome 16 on the Spontaneous Meta
static Ability of AT6.1 Cells. To examine the effect of human
chromosome 16 on tumorigenesis and metastasis, we transferred the
chromosome into a highly metastatic AT6.1 cell line using the MMCT
technique. Approximately 200 clonal cell lines were established, and
24 of them were subjected to screening by PCR. Based on this
preliminary screen, seven hybrid clones containing various regions of
human chromosome 16 were chosen, designated as AT6.1-16-2,
AT6.1-16-6, AT6.1-16-7, AT6.1-16-9. AT6.1-16-10, AT6.1-16-11.
and AT6.1-16-12, and used for further analysis.
±3792
clonesAT6.AT6.AT6.AT6.AT6.AT6.AT6.-16-9-16-12-16-2-16-10-16-6-16-7-16-1115/157/77/75/54/45/512/1210/1
±2380
220.7±
0.820
±
1811
±
±225±
124±3Median'7787820.516013(range)''(24-138)(52-13
" Number of tumors bearing SCID mice/number of tumor-inoculated SCID mice.
'*The number of metastatic lesions on the lungs were macroscopically counted after 4
weeks of s.c. inoculation.
' Number of metaslatic lesions on lungs per SCID mouse.
' Number of metastatic lesions on lungs per group of SCID mice.
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16
press metastasis. On the other hand, expression of the M-cadherin
gene was not detectable in any hybrid clones except AT6.1-16-2.
These results suggest that neither the H- nor M-cadherin gene is
involved in the suppression of tumor metastasis in our system.
Discussion
Fig. 1. FISH analysis of the AT6.1-16-2 cell. Metaphase chromosomes were prepared
from AT6.1-16-2 cells as described in the text and hybridi/ed with the whole chromosome
painting probe lor chromosome 16 {Vysis Co.). Arrowheads, human chromosome 16.
hybrid clones. AT6.1-16-9 and AT6.1-16-12, that still retained metastatic ability to the lungs in SCID mice. This DÌ6S402marker is
located in the q24.2 region of the long arm of chromosome 16 ~ 109
cM from the top of the short arm of the chromosome. These results
suggest that a potential metastasis suppressor gene responsible for the
suppressor activity is located in a proximate area to the q24.2 region
on human chromosome 16. To verify the location of the suppressor
region, we prepared DNA from metastatic lesions in the lungs of mice
injected with one of the hybrid clones. AT6.1-16-2, which suppresses
metastasis, and examined the status of the DÌ6S402marker. We
reasoned that because AT6.1-16-2 cells significantly suppress metas
tasis, those metastatic tumors in the lungs should lose the suppressor
gene by chromosome rearrangement. As shown in Fig. 2G of nine
tested tumor tissue samples from the lung metastatic lesion, we found
that all of them lost the DÌ6S402marker, while D16S539 was retained
in all of the samples. These results strongly support our conclusion
that the candidate suppressor gene is located in close proximaty to the
region of DÌ6S402.
The human chromosome 16q arm is known to contain the E- and
P-cadherin genes that have been reported to be involved in the
progression of a variety of tumors (28, 29); therefore, they are con
sidered to play key roles in suppressing tumor metastasis (28). To
determine whether the function of the E- and P-cadherin genes are
involved in metastasis suppression in our system, PCR analysis was
conducted using a pair of primers for these genes. As shown in Fig.
2/4, the hybrid clone AT6.1-16-7, which showed a significant reduc
tion in metastasis, lost the E-cadherin gene, whereas two other clones
that failed to suppress metastasis (AT6.1-16-9 and AT6.1-16-12) still
retained the E-cadherin gene. Similarly, the P-cadherin gene were
lost from two clones (AT6.1-16-6 and AT6.1-16-7) that are capable of
suppressing metastasis, whereas this gene appeared to be intact in one
clone (AT6.1-16-12) that failed to suppress metastasis. Therefore, the
metastasis suppressor activity shown in our system is not due to either
the E- or P-cadherin gene. Other cadherin genes, i.e., H- and Mcadherin, are recently mapped in the q24.2 and q24.3 regions of
chromosome 16. respectively (30). Because of their close proximity to
our suppressor locus, we examined the expression of these genes in
our hybrid clones by RT-PCR. As shown in Fig. 2D, mRNA of the
H-cadherin gene was not detectable in three clones, AT6.1-16-6,
AT6.1-16-7, and AT6.1-16-11, that are capable of suppressing me
tastasis, whereas two other clones (AT6.1-16-9 and AT6.1 -16-12) still
expressed the H-cadherin gene, although both clones failed to sup
Although the clinical importance of tumor metastasis is well rec
ognized, advances in understanding the molecular mechanism in
volved in metastasis formation have lagged behind other develop
ments in the cancer field. This is because of the fact that metastasis
involves multiple steps with high complexity and is controlled by a
variety of positive and negative factors. However, recent findings
about a series of metastasis suppressor genes shed new light on the
process of tumor metastasis progression (12-20). The MMCT has
been proven to be a particularly useful method to identify such
dominant suppressor genes as an alternative to the conventional
positional cloning approaches. Using the combination of this powerful
technique and a highly metastatic rat prostate cancer model in SCID
mice, we found that the human chromosome 16 has a strong suppres
sor activity of tumor metastasis, whereas it did not affect tumorigenesis itself. A structural analysis in various hybrid clones with STS
markers by PCR revealed that there is a metastasis suppressor gene in
the q24.2 region of chromosome 16.
The genetic abnormalities of human chromosome 16, especially at
its q arm, are most frequently and consistently observed in prostate
cancer. Based on a CGH analysis, Visakorpi et al. (6) reported that as
much as a 19% clinical sample of primary prostate cancer showed a
deletion in the q arm. In another report using the same CGH analysis,
at least 30% of primary prostate cancer (7) and 55% of metastatic
clinical samples were found to have deletions at the q arm of chro
mosome 16 (8). Furthermore, an LOH analysis including 59 samples
from prostate cancer patients provided evidence that there are signif
icantly high incidences of deletions in the region between q22 and qter
of chromosome 16 (3). It is noteworthy that deletions in these regions
are also frequently observed in breast cancer (31, 32). Our PCR
analysis suggested that a metastasis suppressor gene is located in the
q24 region, which coincides with the region in which an allelic
imbalance was most frequently reported in human prostate cancer.
LOH has been reported in clinical samples in the q24.1-q24.2 region
in 76% of aggressive prostate cancer (4) and in the q24.3 region in
80% of metastatic prostate cancer (3). Furthermore, LOH in the region
between q24.1 and its terminus are most frequently, up to 50%,
observed in cancer death cases (2). These results with clinical samples
strongly support our notion that there is a metastasis suppressor gene
in the q24 region of chromosome 16.
There are several genes that are located at the chromosome 16q arm
and are potentially involved in tumor metastasis progression. The
E-cadherin gene, which encodes an adhesion molecule and is located
in the q22 region, was shown to be significantly down-regulated in
high-grade prostate tumors, and the decreased expression of this gene
is associated with the poor prognosis of prostate cancer patients (28).
Similarly, the P-cadherin gene is also located in the q22 region and
has been reported to be involved in suppressive roles of melanoma
progression (29). However, our PCR analysis of microcell hybrid
clones indicated that neither £-nor P-cadherin genes are responsible
for the metastasis suppression in our system. Two other cadherin
genes, i.e.. M- and H-cadherin, are located in the q24 region, and they
are considered to be potentially involved in tumor metastasis (33, 34).
In particular, reduction in H-cadherin expression has been observed
frequently in breast cancer (33). However, our RT-PCR analysis of
hybrid clones suggest that neither genes are involved in the suppres
sion of metastasis in our system. The HSDI7B2 gene located at
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SUPPRESSION
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I'HKOMOSOMK
Id
(A)
MMCT Chl6 clones
Metastasis suppression
STS
Location
Markers
(cM)
D16S521 0
D16S510 8.3
D16S748
11
D16S406
14.8
D16S407
16.7
D16S764 20
D16S500 27
D16S292 28
D16S499 33.3
D16S769 39
D16S403 42.7
D16S420 43.2
D16S401 45.5
_D16S753
46
—¿
D16S409
D16S3253
D16S415
DI6S408
D16S2624
D16S514
D16S3021
E-cadherin
P-cadherin
D16S186
D16S515
D16S3142
D16S516
D16S507
D16S402
D16S539
D16S3037
D16S520
D16S3048
- D16S413
56.2
60
65.6
72.6
76
80
83.1
84
84
85
90.2
92.6
98.3
103.1
109.9
117
119.2
123.3
125.9
126.5
2 106 7 11 9 12
+ + + + + - -
(B)
•¿â€¢â€¢â€¢O
•¿â€¢
•¿OO«O «O
•¿OOOO OO
•¿â€¢OOO•¿â€¢
•¿â€¢â€¢OO
•¿â€¢
•¿â€¢â€¢â€¢O
•¿â€¢
DI6S402
D16S539
•¿â€¢OO«
•¿â€¢
•¿â€¢OOO
•¿â€¢
(C)
•¿â€¢OO*
•¿â€¢
333333ÕC
•¿â€¢â€¢â€¢O
•¿â€¢
•¿â€¢â€¢O«
•¿â€¢
•¿o««o
•¿â€¢
•¿â€¢ooo
«o
•¿â€¢ooo
•¿â€¢
•¿â€¢oo«
•¿â€¢
•¿â€¢â€¢o«
•¿â€¢
•¿â€¢00«
O«
•¿â€¢o««
•¿â€¢
•¿oooo
oo
•¿â€¢oo«
•¿â€¢
•¿â€¢ooo
•¿â€¢
_ _ —¿.
Retained region in
DI6S402
D16S539
(D)
1 ' <->J the suppressed
•¿
•¿ clones
H-cadherin
•¿â€¢000
«O
M-cadherin
•¿0*00
*o
•¿â€¢â€¢â€¢o
•¿â€¢
Beta-actin
•¿Retention of the indicated marker on the transferred
chromosome 16.
O Absence of the indicated marker on the transferred
chromosome 16.
Fig. 2. Structural analysis of chromosome 16 in microcell hybrid clones hy PCR. A. chromosomal DNAs were prepared from five hybrid clones (AT6. 1-16-2. AT6.I-I6-IO,
AT6. 1- 16-6. AT6. 1- 16-7. and AT6. 1-16-11) that were suppressed for their metastasis and also two hybrid clones (AT6. 1-16-9 and AT6. 1-16-12) that retained metastasis abilities. DNAs
were subjected to PCR using various primers of STS markers for human chromosome 16 and also E-cadherin exon 13 primers and P-cadherin exon I primers as described in the text.
•¿.
retentions of the indicated markers; O. absences of the indicated markers of chromosome 16. B, a representative result of PCR analysis for various hybrid clones using STS markers.
D16S402 and DI6S539. C, PCR analysis of tumor tissues from metastatic lesions in the lungs. Nine tumor tissues, designated as LM I through LM9. were removed from the lung
metastatic lesion of mice injected s.c. with AT6. 1-16-2. DNAs were prepared directly from these tissues and subjecled to a PCR analysis using STS markers. Dlf>S402 and l)lf>S5JV.
D, RT-PCR analysis of H- and M-ciiilherin genes. Total RNAs were prepared from various hybrid clones and subjected to a RT-PCR analysis using each of a set of primers for //and M-cddlierin genes. Total RNA of human skeletal muscle was obtained from Clontech Co. and used as a control for RT-PCR.
q24.1-24.2 encodes the 17HSD type 2 enzyme, which converts 17hydroxysteroids to a 17-keto form to inactivate sex steroids in the
prostate, and this enzyme is implicated in prostate carcinogenesis
(35). Both DPEP1 (renal dipeptidase) and BBC! (breast basic con
served gene) genes are also located at q24.3, and they are known to be
down-regulated in Wilms' tumor (36) and breast cancer (37), respec
tively, suggesting their roles in tumor progression. The cell matrix
adhesion regulator gene, CMAR, resides at q24.3 and encodes a signal
transduction molecule, and its roles were implicated in suppression of
tumor invasion (38). The c-myc promoter binding protein gene, which
is a negative regulator of the c-myc gene, is also located at the q24
region, and the loss of this gene can increase the c-myc expression and
may lead to the progression of tumorigenesis (39). Because these
genes have potential roles as suppressors in tumor progression, they
may be involved in the metastasis suppression shown in our test
system. However, the roles of these genes in prostate tumor metastasis
remained to be examined.
In summary, our data represent the first functional evidence of
tumor metastasis suppressor activity on human chromosome 16. A
putative gene of the suppressor is located in the q24.2 region of the
chromosome. Further characterization and cloning of this gene should
be useful to understand prostate tumor progression and to develop a
diagnostic marker for this devastating disease.
References
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273-287. 1997.
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Shimazaki. J. Three distinct commonly deleted regions of chromosome arm loq in
human primary and metastatic prostate cancers. Genes Chromosomes Cancer. 17:
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BY HUMAN CHROMOSOME
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Human Chromosome 16 Suppresses Metastasis But Not
Tumorigenesis in Rat Prostatic Tumor Cells
Tomoyuki Mashimo, Misako Watabe, Andrew P. Cuthbert, et al.
Cancer Res 1998;58:4572-4576.
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