Download Cloning, Characterization, and Chromosomal

ICANCKR RESEARCH 58. 2196-2199.
May 15. 1998]
Cloning, Characterization, and Chromosomal Localization of a Gene Frequently
Deleted in Human Liver Cancer (DLC-1) Homologous to Rat RhoGAP
Bao-Zhu Yuan, Mark J. Miller, Catherine L. Keck, Drazen B. Zimonjic, Snorri S. Thorgeirsson,
Nicholas C. Popescu1
laboratory
of Experimental Cttrcinogenesis,
and
Division of Basic Sciences, National Cancer Institute. NIH. Rethesda. Maryland 20H92
ABSTRACT
The isolation of genes involved in cancer development is critical for
uncovering the molecular basis of cancer. We report here the isolation of
the full-length cDNA and chromosomal localization of a new gene fre
quently deleted in liver cancer (DLC-1> that was identified In represen
tational difference analysis. Loss of heterozygosity was detected for DLC-1
in 7 of 16 primary hcpatocellular carcinomas (HCCs) and in 10 of 11 HCC
cell lines. Although mRNA for DLC-1 was expressed in all normal human
tissues, it was not expressed in 4 of 14 HCC cell lines. Full-length cDNA
for DLC-1 of 380«hp encodes a protein of 1091 amino acids, has 86%
homology with rat pl22 RhoGAP gene, and was localized by fluorescence
in situ hybridization on chromosome 8 at bands p21.3-22. Deletions on the
short arm of chromosome 8 are recurrent in liver, breast, lung, and
prostate cancers, suggesting the presence of tumor suppressor genes.
DI.C-I may be a tumor suppressor gene in liver cancer as well as in other
Qidong Liver Cancer Institute. China (9). and Dr. Curtis C. Harris (Laboratory
of Human Carcinogenesis. Division of Basic Sciences. National Cancer Insti
tute).
RDA. One primary HCC. having a homo/.ygous point mutation of thc/>5.?
gene which was not present in the surrounding noncancerous liver tissue, was
selected for analysis. RDA was performed as originally described with tumor
DNA as tester and normal liver DNA as driver (10). BglU (Promega Corp.,
Madison. WI) was chosen as the restriction enzyme, and its adaptors were used
for direct preparation of amplicons and PCR-based subtractive hybridization.
The final difference products showing distinct bands in agarose gel were
recovered after Bglll digestion and ligated into the ß#/IIsite of dephosphorylated pSP72 vector. The recombinant difference products were then transfected into Eschericlna culi DHIOB.
Characterization of RDA Probes. Plasmids with a distinct DNA insert
were selected for further analysis. DNA sequencing was performed using the
Dye Terminator Cycle DNA Sequencing kit (Perkin-Elmer. Rockville. MD).
Sequencing reaction products were purified by spin columns (Princeton Sep
arations. Adelphia. NJ) and run in a 377 DNA Sequencer (Perkin-Elmer/
INTRODUCTION
Human cancer is viewed as a malady of genes originating in the
progeny of a single cell through accumulation of multiple genetic
alterations that may be caused by diverse types of damage (1-4). A
major challenge in cancer biology is the identification of cancerrelated genes, which comprise only a fraction of the human genes. We
describe here our search for a cancer gene in HCC,2 one of the most
common cancers worldwide (5, 6).
We used RDA, a PCR-based subtractive hybridization technique
(7), which was applied to tissues derived from a primary HCC and
adjacent noncancerous tissue from the same patient. Several amplified
and deleted fragments detected by RDA were isolated. Here, we
present the characterization and cloning of a deleted fragment that
resulted in the identification of a new gene designated DLC-1 highly
homologous to the rat pl22 RhoGAP gene (8). The DLC-I gene was
mapped on chromosome 8p21.3-22, a region frequently deleted in
solid tumors.
MATERIALS
AND METHODS
Primary HCC Samples and HCC Cell Lines. All of the primary tumor
DNAs were obtained from surgical resection of HCC tissues from patients in
Qidong. China. Each tumor sample was matched with its surrounding noncan
cerous liver tissue. DNAs were extracted after diagnosis of HCC with or
without cirrhosis. The tumors were HBV positive for HBV surface antigen
and/or PCR detection of HBV.\ gene. HCC cell lines used in these experiments
were obtained from American Type Culture Collection (Rockville. MD).
Received 1/8/98; accepted 3/18/98.
The costs of publication of this article were defrayed in part 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.
' To whom requests for reprints should be addressed, at National Cancer Institute.
Applied Biosystems. Foster City. CA). The homology analysis was carried out
by BLAST search of the GenBank DNA databases (II). The GenBank acces
sion number for the human DLC-1 sequences in this report is AF035119. The
RDA products, which elicited significant homology or appeared in multiple
clones, were selected for further Southern blot and/or Northern blot analysis.
Software ImageQuant version 3.3 (Molecular Dynamics. Sunnyvale. CA) was
used for quantitative analysis. ß-Actinwas used as a standard in quantitative
Southern blot analysis.
5' and 3' RACE and cDNA Library Screening for cDNA Cloning. 5'
and 3' RACE started from a deleted fragment detected from RDA was
performed using human placenta Marathon cDNA as template (Clontech.
Inc.. Palo Alto. CA). The final _VRACE product, exhibiting the same band
pattern as the deleted fragment in Northern blot hybridization, was labeled
with |a-'2P]dCTP
to screen a 5' stretch cDNA library constructed from
human lung tissue (Clontech, Inc.). The lambda DNA of positive clones
was converted into plasmid DNA by transfecting lambda DNA into AMI
bacterial cells. The full-length cDNA sequencing of positive clones was
completed by primer walking and assembled by the Sequencher
3.1
program.
FISH Gene Mapping and CGH. A genomic probe isolated from a human
PI library was labeled with biotin and used for FISH chromosomal localiza
tion. For CGH analysis, the DNA from primary tumor used as a tester in RDA
was cohybridized with DNA from normal cells. For both analyses, chromo
somes prepared from methotrexate-synchronized
normal peripheral lympho
cyte cultures were used. The original CGH protocol with minor modification
was used ( 12). The conditions of hybridization, the detection of hybridization
signals, digital-image acquisition, processing and analysis, and direct fluores
cent signal localization on banded chromosomes
previously (13).
were performed as described
RESULTS
RDA. Several RDA difference products were observed after the
third round of hybridization/selection as distinct bands in agarose gel.
Six individual fragments were isolated and analyzed by Southern blot
Building 37. Room 3C28. 37 Convent Drive MSC4255. Bethesda. MD 20892-4255.
hybridization for deletions. One clone (L7-3) of 600 bp showed loss
Phone: (301)496-5688; Fax: (301)496-0734.
- The abbreviations used are: HCC, hepatocellular carcinoma; RDA. representational
of heterozygosity in the primary tumor (Fig. IA). A BLAST search
difference analysis: DLC. deleted in liver cancer; HBV. hepatitis B virus; RACE, rapid
revealed that the L7-3 clone had homology to rat p 122 RhoGAP
amplification of cDNA ends; FISH, fluorescence in situ hybridization: CGH. comparative
genornic hybridization.
cDNA.
2196
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Hl MAN KliodM'
(ÃŒKNI-DI-I.ETI-.I) IN I.IVT.R CANC'I-.K
region
of 8pter-qll.2
and gain
on region
of 8q21.1-q24.3
(Fig. 4b).
DISCUSSION
L7-3
0.5kb
Beta-actin
B
5.0kb
i *
9 ico.K
_i
o.
a s
L7-3
Beta-actin «»T-9ttM»t-
1
0.5kb
5.0kb
Fig. I. Genomic deletions of Ihe L7-3 clone in HCC and HCC cell lines. Genomic
DNAs were digested by Bi>n\ and separated by eleclrophoresis in 1% agarose gel. A,
primary tumors 94-25T. 95-03T. and 95-06T showed 50% decrease of DNA intensity as
compared with normal liver tissues. B. cell lines Sk-Hep-1. PLC/PRF/5. WRL. Focus.
HLF. Hep3B. Huh-7. Huh-6. and Chang showed reduction of DNA intensity in different
degrees compared with human normal liver genomic DNA. WRL and Chang are nonlumorigenic cell lines.
RDA was successfully used to identify a new gene DLC-I from
a primary liver tumor. This gene has high homology with p 122 rat
RhoGAP gene (8) and encodes a protein of 1091 amino acids,
which is likely to be the human homologue of rat RhoGAP protein.
The RhoGAP gene encodes a GTPase activating protein that can
catalyze the conversion of the active GTP-bound Rho complex to
the inactive GDP-bound one. The Rho family proteins, a subfamily
of the Ras small GTP binding superfamily. function as important
regulators in the organization of actin cytoskeleton (14). Rho
proteins are also involved in Ras-mediated oncogenic transforma
tion (15). In addition to RhoGAPs, another category of regulator
proteins is guanine nucleotide exchange factors. Whereas guanine
nucleotide exchange factors act as oncogenic proteins to activate
Rho proteins, GAPs may function as tumor suppressors by downregulating Rho proteins (16). Consistent with this notion, the NFÌ
tumor suppressor gene functions as a GAP gene for Ras. A recent
study showed that the GAP domain of p 190. a ubiquitously ex
pressed RhoGAP. could suppress Ras-mediated
tumorigenesis
(17).
Deletion of the DLC-l gene was detected in —¿50%
of primary
HCCs and in a majority of HCC cell lines. Moreover, the DLC-Ì
gene was not expressed in 28% of HCC cell lines. DLC-ÃŒgene
To determine if the L7-3 clone is represented in a region recurrently
deleted in HCCs, we examined 16 primary HCCs and 11 HCCderived cell lines. Seven primary HCC and 10 HCC cell lines had
genomic deletion of L7-3 as compared with normal human liver DNA
(Fig. 1). Representative samples from primary HCCs are shown in
Fig. 1/4. Considering the significant DNA sequence homology of the
L7-3 clone with rat RhoGAP cDNA. its mRNA expression was
examined in both normal human tissues and HCC-derived cell lines.
Analysis of mRNA isolated from several normal human tissues,
including liver, demonstrated that the L7-3 clone hybridized to 7.5 kb
(major) transcript and 4.5 kb (minor) transcript that were detected in
all normal tissues but not in 4 of 14 human HCC-derived cell lines
(Fig. 2).
The cDNA for the clone L7-3 was obtained by 5' RACE and 3'
Kb
9.5
7.5
§
RACE coupled with cDNA library screening, and the gene was
named DLC-1. The full-length cDNA of the DLC-1 gene is 3850
bp long and encodes a protein of 1091 amino acids, which is likely
to be a human counterpart of rat RhoGAP protein. The estimated
molecular weight of DLC-1 protein is 125.000. The untranslated
regions of 5' end and 3' end of the DLC-1 gene are 324 and 250
bp. respectively (Fig. 3).
Chromosomal
Localization. To obtain further information re
garding the DLC-Ì
gene and its possible association with liver cancer,
the chromosomal localization was determined. The majority of metaphases hybridized with a biotin- or digoxigenin-labeled genomic
probe had fluorescent signal at identical sites on both chromatids of
the short arm of chromosome 8. The signal was analyzed in 100
metaphases with both homologues labeled. Fifty metaphases were
examined by imaging of 3,3'-diaminobenzidine
generated and en
hanced G-like banding. The fluorescent signals were distributed
within region 8p21-22 (Fig. 4a). However, more than 50% of dou
blets were at bands 8p21.3-22, the most likely location of the DLC-I
B
L7-3
Beta-Actin
gene.
CGH. To further characterize the region harboring the DLC-l
gene, the primary tumor DNA was analyzed by CGH. The fluo
rescence profile for chromosome 8 demonstrated DNA loss on
4.5
*Ȁ,
Fig. 2. mRNA expression of the DLC-I gene in normal human tissues and in HCC cell
lines. A. DLC-1 gene expressed in all normal tissues tested as a 7.5-kh major transcript and
a 4.5-kb minor transcript. B, mRNA of the DLC-l gene was not detected in WRL, 7703.
Chang, and Focus cell lines. NL is the control sample from normal liver cells.
2197
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HUMAN RhoGAPGENE DELETEDIN LIVERCANCER
MCRKKPDTMI
HDFLDRDAIE
WTFQRDSKRW
RQEVSSVRSL
GSLPSPKELS
SKLGLIISGP
TQTSSSSSQS
NNWEQNFKN
WRTGSFHGPG
YSSSGDLADL
DSVSPCPSSP
VGASLTRSNR
KLTALLEKYT
GQPLPQSIQQ
YEGQSAYDVA
AAIMLLPDEN
TLKRENSSPR
EEMSRCRNSY
EKFKGWVSYS
EQHLWDVDLL
PKGACALLLT
LRGHMPEWYT
LTQIEAKEAC
ALCRRLNTLN
SRLEEFDVFS
SSTGSLPSHA
SFSFSMKGHE
ILQEGMDEEK
ETSSAVSTPS
RESYPEDTVF
HISLRRENSS
ENEDIFPELD
KQIHLDVDND
HRLRWHSFQS
PSNKHGFSWA
AMRYLRNHCL
DMLKQYFRDL
RWLQTLLYF
VMQRKQSLGK
TEQELKPLTL
TSEQAELSYK
DSKVIEILDS
SVDHDRAPW
KSFGHLCAAE
DWLRATGFPQ
KCAVMKLEIS
PKQDLVPGSP
PPSEDAATPR
KTAKSKTRSL
LKQLSCVEIS
PVTRTRSLSA
YIPEDHKPGT
DSPKELKRRN
DILYHVKGMQ
RTTPSDLDST
SHRPSLNSVS
VPKFMKRIKV
DQVGLFKKSG
PEPLMTNKLS
LCDVTAAVKE
PDQKDLNENL
EALGHLGNDD
KVSEGPRLRL
QTEIYQYVQN
GVRVNVLLSR
WKIRDSFSN
YAQLYEDFLF
PHRKRSDDSD
DDSHPKDGPS
TNSVISVCSS
LKRMESLKLK
ALNGNRINVP
CNKRVGMYLE
FPKALTNGSF
SSSSMSSRLS
RIVNQWSEKF
GNSLNEPEEP
LQINCQSVAQ
PDYKDRSVFG
VKSRIQALRQ
ETFLQIYQYV
NQMTPTNLAV
AATQGLAHMI
SADYQHFLQD
WRSVIEVPAV
SMAPHPARDY
YLIEPCGPGK
QNTETKDTKS
PIDISLVKRE
EDEPCAISGK
PGGTLMDLSE
SNLAGNDDSF
SSHHSKHKAP
MVRKRSVSNS
GFDPFNQSTF
SPSGNNGSVN
IYDNVPGSIL
SDEGDSDSAL
SEIPERRDSG
MNLLQKYSLL
VPLTVNVQRT
MNEGAIDCVN
PKDQRLQAIK
CLAPSLFHLN
AECKKLFQVP
CVDGLFKEVK
PEEILKRLLK
WLRTWRTNL
SKLTYMCRVD
R
0050
0100
0150
0200
0250
0300
0350
0400
0450
0500
0550
0600
0650
0700
0750
0800
0850
0900
0950
1000
1050
1100
Fig. 3. Amino acid sequence of the DLC-I gene.
localization on chromosome 8p21.3-22 and CGH profile of chro
mosome 8 of the primary tumor are significant. Loss of heterozygosity is frequently observed at chromosome 8p22-21 in HCC,
non-small cell lung carcinoma, colorectal cancer (18), and prostate
cancer (19-21). A physical map of contiguous cosmid markers in
Fig. 4. a. chromosomal locali/ation of the
DLC-I gene hy FISH. Digital image of a normal
human mctaphase exhihiting G-like banding of
3,3'-diaminohen/idine-counterstained
chromo
somes. Both chromosomes 8 have fluorescent sig
nal on the short arm at band 8p21.3-22. b, CGH
analysis. CGH analysis of chromosome 8 showing
(from left to righi ) a pair of G-banded chromosome
8, the same chromosomes after hybridi/ation with
tumor and normal DNA, and average fluorescence
profile generated from the analysis of 35 metaphases. Red and green bars next to the ideogram
of chromosome 8 indicate the regions of DNA loss
and gain.
8p22-21.3 was constructed by FISH and pulsed-field electrophoresis. Based on this map, a 1.2-Mb region between markers cC18245 and cC18-2644/cMCR was found to be deleted in HCC,
colorectal cancer, and non-small cell lung carcinoma, suggesting
the presence of a tumor suppressor gene(s) in this region common
to these types of cancers. Furthermore, a 600-kb fragment from the
region commonly deleted in these malignancies was isolated (22,
23). The position of the DLC-Ì
gene relative to the yeast artificial
chromosome contigs for 8p22-21.3 as well as a more specific
mapping of the gene in the radiation-reduced DNA panels remains
to be determined. This refined mapping will show whether the
DLC-Ì
gene is within the region of recurrent deletions and hence
is a strong candidate tumor suppressor gene. Meanwhile, a CGH
profile of chromosome 8 revealed DNA loss on the short arm and
gain on the long arm. This imbalance was highlighted for the first
time in prostate cancer (24) and recently detected by CGH in
primary HCC, non-small cell lung carcinoma, and node-negative
breast carcinomas (25-27). Recurrent chromosome 8 imbalances in
several solid tumors strongly support the relevance of this lesion in
the pathogenesis of human cancer. The location of the DLC-I gene
on a chromosomal region of frequent deletions suggests that this
's/*«
u '
r¡
2198
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HUMAN RhuGAP GENE DELETED
gene is a candidate tumor suppressor gene for human liver as well
as for prostate, lung, and breast cancer.
ACKNOWLEDGMENTS
We thank Dr. Eugenio Santos for valuable discussions on the RhoGAP gene
function.
REFERENCES
1. Bishop. J. M. The molecular genetics of
305-311, 1987.
2. Croce, C. M., and Nowell, P. C. Molecular
65: 1-7, 1985.
3. Fearon, E. R., and Vogelstein. B. A genetic
61: 759-767, 1990.
4. Knudson, A. G. Antioncogenes and human
10914-10121. 1993.
5. Simonetti, R. G., Camma, C., Fiorello, F.,
6.
7.
8.
9.
10.
U.
12.
13.
14.
cancer. Science (Washington
DC), 235:
basis of human B cell neoplasia. Blood.
model for colorectal tumorigenesis. Cell,
cancer. Proc. Nati. Acad. Sci. USA, 90:
Politi. F., D'Amico. G., and Pagliaro, L.
Hepatocellular carcinoma. A worldwide problem and the major risk factors. Dig. Dis.
Sci., 36: 962-972, 1991.
Harris, C. C. Hepatocellular carcinogenesis: recent advances and speculations. Cancer
Cells, 2: 146-148, 1990.
Lisitsyn, N., Lisitsyn, N., and Wigler, M. Cloning the differences between two
complex genomes. Science (Washington DC), 259: 946-951, 1993.
Homma, Y., and Emori, Y. A dual functional signal mediator showing RhoGAP and
phospholipase CS stimulating activities. EMBO ].. 14: 286-291. 1995.
Wang, G.. Zhu. D.. Ye. X., and Chen, R. Establishment and characterization of a
tumor cell line, QGY-7703. derived from hepatocellular carcinoma of a Qidong
patient. Chin. J. Oncol.. 3: 241-244, 1981.
Lisitsyn, N. A., Lisitsina, N. M.. Dalbagni. G.. Barker. P.. Sanchez, C. A., Gnarra. J.,
Linehan, W. M., Reid. B. J., and Wigler. M. H. Comparative genomic analysis of
tumors: detection of DNA losses and amplification. Proc. Nati. Acad. Sci. USA, 92:
151-155, 1995.
Altschul, S. F., Gish. W., Miller, W.. Myers, E. W., and Lipman. D. J. Basic local
alignment search tool. J. Mol. Biol.. 2/5: 403-410. 1990.
Kallioniemi, A., Kallioniemi, O. P.. Sudar, D.. Rutovitz, D., Gray, J. W., Waldman.
F.. and Pinkel. D. Comparative genomic hybridization for molecular cytogenetic
analysis of solid tumors. Science (Washington DC), 258: 818-821. 1992.
Zimonjic, D. B., Rezanka, L., DiPaolo, J. A., and Popescu. N. C. Refined localization
of the erbB-3 proto-oncogene by direct visualization of FISH signals on LUT-inverted
and contrast-enhanced digital images of DAPl-banded chromosomes. Cancer Genet.
Cytogenet., 80: 100-102, 1995.
Nobes, C. D., and Hall, A. Rho, rac, and cdc42 GTPases regulate the assembly of
multimolecular focal complexes associated with actin stress fibers, lamellipodia. and
filopodia. Cell, 81: 53-62, 1995.
IN LIVER CANCER
15. Khosravi-Far. R , Campbell. S., Rossman. K. L.. and Der. C. J. Increasing complexity
of Ras signal transduction: involvement of Rho family proleins. Adv. Cancer Res.,
69: 59-105. 1997.
16. Quilliam, L. A.. Khosravi-Far, R.. Huff, S. Y.. and Der, C. J. Guanine nucleotide
exchange factors: activators of the Ras superfamily of proteins. Bioessays, 17:
395-404, 1995.
17. Wang, D. Z., Nur, E. K. M. S., Tikoo, A., Montague, W., and Manila, H. The GTPasc
and Rho GAP domains of p 190, a tumor suppressor protein that binds the M(r)
120,000 Ras GAP. independently function as anti-Ras tumor suppressors. Cancer
Res., 57: 2478-2484, 1997.
18. Emi, M., Fujiwara. Y.. Ohata, H.. Tsuda, H., Hirohashi. S.. Koike, M.. Miyaki, M.,
Monden. M., and Nakamura. Y. Allelic loss at chromosome band 8p2l.3-p22 is
associated with progression of hepatocellular carcinoma. Genes Chromosomes Can
cer. 7: 152-157, 1993.
19. Visakorpi. T.. Kallioniemi. A. H.. Syvanen. A. C.. Hyytinen. E. R.. Karhu, R.,
Tammela, T., Isola. J. J.. and Kallioniemi, O. P. Genetic changes in primary and
recurrent prostate cancer by comparative genomic hybridization. Cancer Res., 55:
342-347. 1995.
20. Bergerheim. U. S.. Kunimi. K., Collins, V. P., and Ekman, P. Deletion mapping of
chromosomes 8. 10, and 16 in human prostatic carcinoma. Genes Chromosomes
Cancer, 3: 215-220. 1991.
21. Cher. M. L.. MacGrogan. D.. Bookstein. R.. Brown. J. A., Jenkins, R. B., and Jensen.
R. H. Comparative genomic hybridization, allelic imbalance, and fluorescence in situ
hybridization on chromosome 8 in prostate cancer. Genes Chromosomes Cancer. II:
153-162, 1994.
22. Fujiwara, Y., Ohata. H., Emi, M.. Okui, K., Koyama, K., Tsuchiya, E., Nakajima, T.,
Monden. M.. Mori. T.. Kurimasa. A.. Oshimura. M., and Nakamura. Y. A 3-Mb
physical map of the chromosome region 8p21.3—>p22. including a 600kb region
commonly deleted in human hepatocellular carcinoma, colorectal cancer, and nonsmall cell lung cancer. Genes Chromosomes Cancer, lu: 7-14, 1994.
23. Chinen. K., Isomura. M.. Izawa. K.. Fujiwara. Y.. Ohata. H.. Iwamasa. T., and
Nakamura, Y. Isolation of 45 exon-like fragments from 8p22-p21.3, a region that is
commonly deleted in hepatocellular. colorectal. and non-small cell lung carcinomas.
Cytogenet. Cell Genet.. 75: 190-196. 1996.
24. Brothman, A. R. Cytogenetic studies in prostate cancer: are we making progress?
Cancer Genet. Cytogenet.. 95: 116-121. 1997.
25. Petersen, I.. Bujard. M.. Petersen. S.. Wolf. G.. Goeze, A.. Schwendel, A.. Langreck.
H.. Gellen. K.. Reiche!. M.. Just, K., du Manoir. S.. Cremer. T.. Dietel. M., and Ried,
T. Patterns of chromosomal imbalances in adenocarcinoma and squamous cell car
cinoma of the lung. Cancer Res.. 57: 2331-2335, 1997.
26. Isola, J. J., Kallioniemi. O. P.. Chu. L. W., Fuqua. S. A.. Hilsenbeck. S. G., Oshorne,
C. K.. and Waldman. F. M. Genetic aberrations detected by comparative genomic
hybridization predict outcome in node-negative breast cancer. Am. J. Pathol., 147:
905-911, 1995.
27. Marchio. A., Meddeb. M.. Pineau, P.. Danglot. G., Tiollais. P., Bemheim, A., and
Dejean. A. Recurrent chromosomal abnormalities in hcpatocellular carcinoma de
tected by comparative genomic hybridization. Genes Chromosomes Cancer, Iti:
59-65. 1997.
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Cloning, Characterization, and Chromosomal Localization of a
Gene Frequently Deleted in Human Liver Cancer ( DLC-1)
Homologous to Rat RhoGAP
Bao-Zhu Yuan, Mark J. Miller, Catherine L. Keck, et al.
Cancer Res 1998;58:2196-2199.
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