[CANCER RESEARCH 51, 5817-5820, November 1, 1991]
`
`Common Regions of Deletion on Chromosomes 5q, 6q, and lOq in Renal
`Cell Carcinoma
`
`Ryoji Morita, Susumu Saito, Jiro Ishikawa, Osamu Ogawa, Osamu Yoshida, Kazuhiro Yamakawa,
`and Yusuke Nakamura1
`
`Department of Biochemistry, Cancer Institute, Kami-ikebukuro, Toshima-ku, Tokyo 170, Japan ¡R.M., S. S., K. Y., Y. N.J; Department of Genetics Research Laboratory,
`SRL, Inc., Komiya-machi, Hachioji City, Tokyo 192, Japan ¡R.M., S. S.J; Center for Biotechnology, Baylor College of Medicine, Woodlands, Texas 77381 ¡J.I.]; and
`Department of Urology, Faculty of Medicine, Kyoto University, Shogoin-kawahara-cho 54, Sakyo-ku, Kyoto 606, Japan [O. O., O. Y.]
`
`ABSTRACT
`
`5q, 6q,
`on chromosomes
`losses of heterozygosity
`frequent
`Relatively
`on the short arm of
`and lOq,
`in addition to loss of heterozygosity
`chromosome
`3, have been observed in renal cell carcinomas. As the first
`step toward isolation of tumor suppressor
`genes on these three chromo
`somal
`arms, we used six restriction
`fragment
`length polymorphism
`markers for 5q, nine for 6q, and eight for lOq to identify regions commonly-
`
`deleted in a panel of 64 renal cell carcinomas. Allelic losses were common
`at chromosome
`5q21,
`the region where the MCC (mutated in colorectal
`cancer) gene was recently identified; at chromosome
`6q27; and at chro
`mosome
`10q21-23.
`Furthermore,
`an association was observed between
`accumulation
`of allelic
`losses
`on these
`three chromosomal
`arms and
`progression of tumors. Loss of heterozygosity
`on chromosome S showed
`a correlation with the histopathological
`grade of a given tumor and the
`incidence of distant metastasis.
`
`INTRODUCTION
`to investigate the
`Numerous
`studies using RFLP2 markers
`molecular biology of carcinogenesis
`have suggested that an
`accumulation
`of genetic alterations
`at specific chromosomal
`loci contributes
`to tumor development
`and/or
`progression.
`Several genes,
`including
`oncogenes
`and tumor
`suppressor
`genes, appear
`to be involved in tumorigenesis
`of colon and
`breast cancers (1-3).
`In RCC, several lines of evidence point
`to
`the existence of a tumor suppressor gene(s) on the short arm of
`chromosome
`3: somatic
`chromosomal
`aberrations
`involving
`this chromosome
`(4, 5); t(3;8) balanced translocations
`in germ-
`line cells of some patients with hereditary renal cell carcinoma
`(6); and frequent LOH on chromosome 3p (7-10). Our earlier
`study suggested that,
`in addition to the gene on chromosome
`3p, tumor suppressor genes associated with RCC may exist on
`chromosomes
`5q, 6q,
`lOq,
`llq,
`and 17p (11). However,
`the
`frequency of LOH on these chromosomal
`arms was not as high
`as that on chromosome
`3p. To examine further whether
`these
`chromosomal
`arms contain tumor suppressor genes, and if they
`do, to map the loci precisely, we have analyzed these candidate
`loci in detail with a large number of RFLP markers. Addition
`ally, we have expanded the number of tumors
`investigated to
`64 and correlated the accumulation of events causing LOH on
`various chromosomal
`arms with clinical and histopathological
`parameters.
`
`MATERIALS AND METHODS
`
`from 64 patients with
`tissues
`and cancerous
`Materials. Normal
`primary sporadic RCC were obtained during radical nephrectomy at
`
`Received 6/19/91 ; accepted 8/13/91.
`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.
`of
`' To whom requests
`for
`reprints
`should be addressed,
`at Department
`Biochemistry, Cancer Institute, 1-37-1, Kami-ikebukuro. Toshima-ku, Tokyo 170,
`Japan.
`used are: RFLP,
`2The abbreviations
`phism; RCC, renal cell carcinoma: LOH,
`
`fragment
`restriction
`loss of heterozygosity.
`
`length polymor
`
`the School of Medicine, Kobe University,
`the Kobe National Hospital,
`or the Department
`of Urology, Kyoto University. The tissues were
`frozen immediately after nephrectomy and stored at -70°C until ex
`traction of DNA.
`DNA Extraction and Probes. The extraction of DNA from tissues
`was carried out according to methods described previously (3). All
`probes used in this study are listed in Table 1. The probes in the cCI6
`series were isolated from a human/mouse
`hybrid cell line which con
`tained only chromosome 6 as human-derived material
`(12) and were
`mapped by fluorescent
`in situ hybridization.'
`Southern Blotting and Hybridization. All DNA samples were trans
`ferred to nylon membrane (Pall Biodyne) with 0.1 N NaOH/0.1 M
`NaCl. Neutralization,
`fixation, and hybridization were done according
`to methods described previously (3).
`
`RESULTS
`
`the enzyme detecting RFLP, and
`localization,
`Chromosomal
`frequency of LOH for each probe are summarized in Table 1.
`Fig. 1 presents examples of deletion maps and Southern blots
`which show loss or retention of heterozygosity
`in tumors. A
`significant
`reduction of signal
`intensity was observed in one of
`two polymorphic
`alÃ(cid:173)elesin blots probed with CI6-39, CI6-49,
`and OS-2. On the other hand, polymorphic alÃ(cid:173)elesin blots with
`CI6-4, THH54,
`and MCT122.2 were retained in tumors. The
`results of LOH studies on four chromosomal
`arms (3p, 5q, 6q,
`and lOq) and the histopathological
`features of each tumor are
`summarized in Table 2. LOH was observed on chromosome
`3
`in 52 of 58 informative cases (90%). LOH also occurred fre
`quently on chromosomes
`5q (13 of 39, 33%), 6q (17 of 44,
`39%), and lOq (25 of 61, 41%). However, LOH on chromo
`somes llq (3 of 26, 12%), 17p (13 of 63, 21%), and 19p (8 of
`55, 15%) was not observed as frequently as in our previous
`study.
`deletion maps of
`In view of these results, we constructed
`chromosomes
`5q, 6q, and lOq. On chromosome
`5q, Tumor 5
`showed LOH with RFLP marker pL5.71-3, but retained both
`alÃ(cid:173)elesat proximal
`loci defined by E5.55 and LS5.3. Tumors
`35 and 94 lost one alÃ(cid:173)eleat
`the L5.71 locus but
`retained
`heterozygosity at the distal YN5.48 locus. From these results,
`a common region of deletion was identified at chromosome
`5q21, where the gene responsible
`for adenomatous
`polyposis
`coli
`is located (Fig. 2a). On chromosome
`6q (Fig. 2b), a
`commonly deleted region was identified at q27:
`the tumor of
`Patient 22 lost an alÃ(cid:173)elein the region distal
`to cCI6-52; Tumor
`33 showed LOH in the region distal
`to cCI6-91; and Tumor
`104 lost a part distal
`to cCI6-4.
`In Fig. 2c, a deletion map of
`chromosome
`lOq shows that Tumor 6 lost an alÃ(cid:173)eleat cTBQ16
`but retained both alÃ(cid:173)elesat pEFD75, Tumor 90 showed LOH
`at THH54
`but
`retained both OS-2 alÃ(cid:173)eles,and Tumor
`104
`showed LOH at OS-2 but retained heterozygosity at THH54
`and MCT122.2. Therefore, a common region of deletion was
`
`3S. Saito et al., unpublished data.
`
`5817
`
`West-Ward Exhibit 1099
`Morita 1991
`Page 001
`
`

`

`COMMON REGIONS OF DELETION ON CHROMOSOMES
`
`5q, 6q, AND I0<j IN RCC
`
`10q
`
`104 on
`Fig. 1. Deletion maps of Tumor
`chromosomes 6q and lOq and typical Southern
`blots demonstrating
`loss or retention of het-
`erozygosity in DNAs from normal kidney (A1)
`and from renal cell carcinoma (T). Al, A2. and
`C indicate alÃ(cid:173)eleI (larger fragment), alÃ(cid:173)ele2
`(smaller fragment), and invariant band, respec
`tively. •¿(cid:3).LOH; D. retained; B. uninformative.
`
`N T
`
`•¿(cid:3)«-A1
`•¿(cid:3)•¿(cid:3)«-A2
`
`«-A1
`
`«-A2
`
`6q
`
`104
`
`CI6-52
`
`CI6-4
`
`CI6-91
`
`CI6-13
`
`CI6-39
`
`CI6-49
`
`JCZ30
`
`
`
`Table 1 Probes tested for loss ofhelerozygosilyProbeChromosome
`
`5
`E5.S5L5.71-3YN5.48-4LS5.3-4
`
`MC5.61KK5.19Total
`
`(chromosome5q)Chromosome
`
`6
`HHH157CI6-7CI6-47
`
`CI6-37
`C16-52CI6-3CI6-4
`
`D6S154
`q21
`D6S164D6S135D6S136
`q23q24q25
`
`(22)
`3/15(20)
`Taql
`1/11(9)5/9
`Msp\Msp\Mspl
`(56)2/16(13)
`
`the q21-q23 band of
`
`between THH54 and OS-2 at
`chromosome
`10.
`between
`
`
`to find correlationsWe attemptedchromosomalalterations
`3,the
`and histopathological
`features. As shown in Fig.
`
`accumulated number of LOHs on chromosomes 3p, 5q,6q,and
`lOq in each tumor showed a correlation with its histopath
`
`ological grade. In the clear cell type of RCC, LOH wasdetectedat
`(17)"8/26(31)5/28(18)3/16(19)
`
`two or more of these chromosomalofmoderately
`arms tested in 82%
`detectedat
`differentiated tumors, although LOH was
`
`only one chromosomalthewell-differentiated
`arm or not at all in nearly half of
`D5S84D6S29D6S138D6S160
`q21q32P21p21.3p21.2
`2/5(40)2/10(20)13/39(33)3/22(14)6/34(18)5/23
`Taq\Rsal//«millPstlBgl\\
`types. The results suggest
`that an accumu
`lation of events causing LOH (probably reflecting loss of func
`tion of tumor suppressor genes) is associated with tumor pro
`gression.
`In particular, LOH on 5q showed a significant corre
`lation with tumor grade (Fisher's
`test, P —¿(cid:3)0.015) and a weak
`association with distant metastasis (P = 0.062) (Table 3). How
`
`ever, stage and tumor size were not correlated with thenumberof
`
`chromosomalLOHs.DISCI
`arms showing
`
`CI6-91CI6-13CI6-39CI6-49JCZ30Total
`D6S186D6S142D6S156D6S161D6S37D10S28
`q26q27q27q27027M**
`7/34(21)8/30
`Msp\Msp]Taq\Taq\Taa\*
`(27)7/27
`(26)6/27
`ISSIONL/lkJ\_-
`(22)9/25
`(36)17/44(39)3/31
`geneson
`lines of evidence have suggested that one or more
`the short arm of chromosome
`3 are associated with devel
`(10)
`
`opment and/or progressionofheterozygosity
`of renal cell carcinomas:
`loss
`
`1/11(9)2/13(15)3/10(30)
`Msp\MsplMspl
`(7-10); cytogenetic aberrations
`(4, 5) and hered
`translocations
`(6); or von Hippel-Lindau
`disease
`itary t(3;8)
`3/15(20)
`
`( 13, 14). In a previous study, we detected two commonlydeletedregions
`5/17(29)9/32
`at chromosomes 3pl4 and 3p21.3 in RCCs (10). How
`(28)8/25
`(32)
`
`ever, studies on colorectal carcinoma(15,16),
`(1, 2), lung cancer
`q22-q25q26q26ql2-ql3.2ql2-ql3.2«13p
`D10S27D10S36D10S25PYGMD11S146INT2D17S5
`3/26(12)5/27(19)10/30(33)25/61
`MsplTaq\TaqlMsplTaqlTaqlTaql
`and breast cancerthatalterations
`
`(Ref. 3; Footnote 4) have shown
`in oncogenes and tumor suppressor genes accumu
`(41)2/15(13)1/15(7)3/9
`
`late to transform a normalthepresent
`cell
`to a malignant
`cell.
`In
`
`study, we have investigatedfivechromosomal
`additional
`loci on
`wereimplicated
`arms
`(5q, 6q,
`lOq, 17p, and 19q) that
`
`by LOH in our previous studies (1 1) to examinethesignificance
`of the allelic losses at
`these loci. Although chro
`13.3
`
`mosomes 17p and 19q did not show the frequency ofLOHreported
`pl3.1-pll.2P13p
`5/28(18)4/20
`D17S31P53D19S21D19S20Locationq21q21q21q21
`MsplBamHlPstlTaqlMsplLOH/informativecases1/6
`to41%
`previously,
`the other
`three regions
`revealed 33%
`(20)13/63(21)3/30(10)8/40
`
`LOH, and commonly deleted regions were identifiedat5q21,6q27,
`and10q21-q23.Kovacs
`
`and Kung (17) and Presti et al. (18) reported cytoge
`(20)7/48(15)8/55(15)assigned
`netic abnormalities on chromosome 5 due to unbalanced trans-
`13.3EnzymeTaqlMsplMsplMsp\
`(chromosome 19p)SymbolD5S133D5SI41D5S81D5S140
`1Numbers in parentheses, percentage.
`
`/pter-pl3
`
`
`
`"f•¿(cid:3)Mspl
`
`
`
`UOOlV/1^1Many
`
`(chromosome6q)Chromosome
`
`10TBQ7
`
`MHZ15TBQ16TB10.163
`
`
`pter-p 13qllq21
`D10S17D10S30D10S22
`
`CMMI7.4
`THH54HHH105OS-2
`
`Pvull
`D10S23
`q21q21q22-q25q22-q25
`D10S14D10SI3D10S20
`MsplB
`filliHindlll
`
`TBQ4MCT
`122.2EFD75Total
`
`(chromosome
`
`Chromosome 11MCMPIHBIS9SS6
`
`lOq)
`
`Total (chromosome 1Iq)Chromosome
`
`17YNZ22
`
`MCT35.1BHP53Total
`
`(chromosome17p)Chromosome
`
`19
`MCOB5JCZ3.1YNZ21Total
`
`(33)
`3/26(12)8/48(17)
`
`13.3pl3.3p
`
`4T. Sato et al., unpublished data.
`
`5818
`
`West-Ward Exhibit 1099
`Morita 1991
`Page 002
`
`

`

`COMMON REGIONS OF DELETION ON CHROMOSOMES
`
`5q. 6q. AND lOq IN RCC
`
`Table 2 (jenolype and histopathological features of tumors
`armPatient
`Genotype of each
`
`3p
`
`type*O
`
`Grade"
`
`5q3
`rII
`014
`—¿(cid:3)15
`019
`02122
`
`026
`—¿(cid:3)29
`032
`034
`037
`•¿(cid:3)51
`053
`054
`•¿(cid:3)5786
`
`•¿(cid:3)8788
`
`091
`092
`094
`•¿(cid:3)95
`097100
`
`0104
`01055
`
`•¿(cid:3)6
`07
`016
`—¿(cid:3)17
`•¿(cid:3)18
`•¿(cid:3)20
`—¿(cid:3)23
`•¿(cid:3)2425
`
`•¿(cid:3)33
`035
`•¿(cid:3)38
`039
`•¿(cid:3)43
`•¿(cid:3)834155
`
`—¿(cid:3)89
`O96
`—¿(cid:3)101
`•¿(cid:3)36
`«040
`0082
`—¿(cid:3)84
`—¿(cid:3)85
`02
`08l
`—¿(cid:3)103
`—¿(cid:3)102
`—¿(cid:3)90
`099
`-98
`—¿(cid:3)52
`—¿(cid:3)5658
`
`—¿(cid:3)59
`
`C•
`C0
`CO
`CO
`C•
`C0
`CO
`C0
`C•
`C—
`C0
`C•
`CO
`C•
`C•
`C•
`CC•
`
`C0
`C0
`C0
`C0
`C0
`C•
`C•
`C0
`CO
`C•
`C0
`C•
`C•
`C0
`CO
`CO
`C•
`C•
`C0
`C0
`C0
`C•
`
`2
`2
`2
`2
`2
`2
`2
`2
`2
`2
`2
`2
`2
`2C2
`
`2
`3
`X
`1
`1
`1
`2
`2
`2
`2
`2
`3
`1
`1
`2
`1
`2
`3
`
`0
`
`•¿(cid:3)
`—¿(cid:3)
`•¿(cid:3)
`•¿(cid:3)
`0
`•¿(cid:3)
`•¿(cid:3)
`
`-
`•¿(cid:3)
`•¿(cid:3)
`
`C•
`C•
`CO
`C0
`GO
`G•
`G•
`GO
`GO
`G•
`G0
`GO
`S0
`C/G0
`C/G0
`C/G•
`C/G•
`C/G0
`G/S•
`XXO
`XO
`X
`•¿(cid:3)
`X
`X•
`MI2
`XX
`»06q&0__O—0•0———00•0•0—o0••0—o•00——0•——_—••——•o——•0o•—••••0—00—•0o00oCelilOq
`
`N=27
`
`N=17
`
`90
`of LOH
`39
`41
`" Grade of each tumor was determined according to the criteria of the Japanese
`Pathological Society (20).
`* C, clear cell type; G. granular cell type; S, spindle cell type; C/G, mixed type
`of clear and granular: G/S, mixed type of granular and spindle: X. not determined.
`' LOH was observed for at least one locus on this arm.
`d Uninformative at every locus on this arm.
`' Retention of hctero/ygosity at every locus on this arm.
`
`M<2
`
`3p and 5q with the breakpoints
`location between chromosomes
`at Bands 3p 13 and 5q22, resulting in an extra copy of the 5q22-
`qter region. It is likely that some of our cases may have partial
`trisomy of 5q. However,
`it was very hard to judge such partial
`trisomy due to difficulties of distinguishing gain of alÃ(cid:173)elefrom
`
`Grade
`Grade
`2or3
`1
`between tumor grade and the number of accumulated
`Fig. 3. Correlation
`mutations detected on four specific chromosomal
`arms (3p. 5q. 6q. and lOq) in
`clear cell carcinomas. M. number of mutations on these arms. Grade I, well
`differentiated: Grades 2 or 3. moderately differentiated.
`
`5819
`
`loss of alÃ(cid:173)eleon Southern blots. We have indicated that a gene
`on chromosome
`5q may be associated with progression
`of
`RCCs. As the commonly deleted region at chromosome
`5q
`includes
`the L5.71 locus, which is a genomic clone of the
`recently ascertained MCC (mutated in colorectal cancer) gene
`(19), the MCC gene might be a candidate for a role in RCC.
`
`05
`
`E
`D
`
`104
`
`D
`E
`
`35
`
`94
`
`D
`D
`El
`D
`
`82
`
`BBD H
`
`D
`E
`D
`B
`
`ED
`
`5D
`
`22
`
`D0B
`
`I0q
`
`TBQI6
`TB10I63
`CMM174
`
`52
`
`90
`
`104
`
`16
`
`D
`
`E3
`
`D BD
`
`DB0
`
`DD
`
`DD
`
`THH54
`HHHI05
`OS-2
`
`21
`
`24
`
`25 Ik
`
`TBQ4
`MCI1222
`EFD75
`
`D
`Fig. 2. Deletion maps of 5q (a), 6q (ft), and lOq (r) in sporadic renal cell
`carcinoma. Common regions of deletion are indicated by a vertical bar on the
`
`right. •¿(cid:3)l.OH: D. retained: D uninformative.
`
`West-Ward Exhibit 1099
`Morita 1991
`Page 003
`
`

`

`COMMON REGIONS OF DELETION ON CHROMOSOMES
`
`5q. 6q. AND lOq IN RCC
`
`Table 3 Correlation between mutation on 5q and tumor grade or distant
`metastasis
`Histopathological
`
`metastasis*-167+4
`
`grade"Retained
`
`LOH11542
`°P= 0.015 (Fisher's test).
`" P= 0.062 (Fisher's test).
`
`or34
`
`8Distant
`
`(1, 2)
`carcinoma
`as in the case of colorectal
`Furthermore,
`and breast cancer (Ref. 3; Footnote 4), accumulation of genetic
`alterations was correlated with the grade of primary renal
`carcinomas. Our results suggested that alterations
`on 6q and
`lOq also are important
`to progression of RCC. Tumors
`from
`Patients 17, 25, 39, 40, and 101 retained heterozygosity at loci
`on chromosome
`3p, but they did show LOH on other chromo
`somal arms. These tumors may contain a very small interstitial
`deletion on chromosome
`3p which was not detectable by our
`panel of probes;
`it is also possible that an alternative pathway
`exists for development of RCC, without association of genes
`on 3p.
`
`REFERENCES
`
`tumorigenesis.
`
`1. Vogelstein, B.. Fearon, E. R., Kern. S. E.. Hamilton, S. R., Preisinger, A.
`C., Nakamura, Y., and White, R. Allelotype of colorectal carcinomas. Science
`(Washington DC), 244: 207-211, 1989.
`2. Fearon, E. R., Vogelstein, B. A genetic model for colorectal
`Cell, 61: 759-767, 1990.
`3. Sato, T., Tanigami, A., Yamakawa, K., Akiyama, F.. Kasumi, F., Sakamoto,
`G., and Nakamura, Y. Allelotype of breast cancer: cumulative alÃ(cid:173)elelosses
`promote tumor progression in primary breast cancer. Cancer Res., 50: 7184-
`7189, 1990.
`4. Yoshida, M. A., Ohyashiki, K.. Ochi. H., Gibas, Z., Pontes, J. E., Prout, G.
`R., Jr., and Sandberg, A. A. Cytogenetic studies of tumor
`tissue from a
`patient with nonfamilial
`renal cell carcinoma. Cancer Res., 46: 2139-2147,
`1986.
`5. Kovacs, G., Szucs, S., DeRiese. W., and Baumgartel, H. Specific chromosome
`aberration in human renal cell carcinoma. Cancer Res.. 40:171-178,
`1987.
`6. Cohen, A. J., Li, F. P., Berg, S., Marchetto, D. T., Tsai, S., Jacobs, S. C,
`and Brown, R. S. Hereditär)' renal cell carcinoma associated with a chro
`mosomal
`translocation. N. Engl. J. Med., 301: 592-595. 1979.
`
`7. Zbar, B., Brauch. H., Talmadge, C., and Lineham, M. Loss of alÃ(cid:173)elesof loci
`on the short arm of chromosome 3 in renal cell carcinoma. Nature (Lond.),
`327:721-724,
`1987.
`8. Kovacs, G.. Erlandsson, R.. Boldog, F., Ingvarsson. S., Muller-Brechlin, R.,
`Klein, G.. and Sumegi, J. Consistent chromosome 3p deletion and loss of
`heterozygosity in renal cell carcinoma. Proc. Nati. Acad. Sci. USA, 85:1571-
`1575, 1988.
`9. Bergerheim, U., Nordenskjold, M., and Collins, V. P. Deletion mapping in
`renal cell carcinoma. Cancer Res., 49: 1390-1396, 1989.
`J., and
`Ishikawa,
`10. Yamakawa, K., Morita. R., Takahashi, E., Hori, T.,
`Nakamura, Y. A detailed deletion mapping of the short arm of chromosome
`3 in sporadic renal cell carcinoma. Cancer Res., 51: 4707-4711,
`1991.
`11. Morita, R., Ishikawa, J., Tsutsumi, M., Hikiji, K., Tsukada, Y., Kamidono,
`S.. Maeda, S., and Nakamura. Y. Allelotype of renal cell carcinoma. Cancer
`Res., 51: 820-823. 1991.
`12. Koi, M.. Shimizu, M.. Morita, H., Yamada, H., and Oshimura, M. Construc
`tion of mouse A9 clones containing a single human chromosome tagged with
`neomycin-resistance gene via microcell fusion. Jpn. J. Cancer Res., 80: 413-
`418, 1989.
`13. King, C. R.. Schimke, R. N., Arthur, T., Davoren, B., and Collins, D.
`Proximal 3p deletion in renal cell carcinoma cells from a patient with von
`Hippel-Lindau disease. Cancer Genet. Cytogenet., 27: 345-348, 1987.
`14. Seizinger, B. R.. Rouleau, G. A., Ozelius, L. J., Laue, A. H., Farmer, G. E.,
`Lamiell, J. M.. Haines, J., Yuen, J. W. M., Collins, D., Majoor-Krakauer,
`D., Bonner. T.. Mathe».C., Runenstein. A., Halperin, J., McConkie-Rosell,
`A., Green, J. S., Trofatter,
`J. A., Ponder, B. A., Eierman, L., Bowmer, M.
`S., Schimke, R.. Oostra, B., Aronin. N.. Smith, D. L, Drabkin, H.. Waziri,
`M. H., Hobbs, W. J., Martuza, R. L., Conneally, P. M., Hsia, Y. E., and
`Gusella. J. F. Von Hippel-Lindau disease maps to the region of chromosome
`3 associated with renal cell carcinomas. Nature (Lond.), 332: 268-269, 1988.
`15. Naylor, S. L., Johnson, B. E.. Minna, J. D., and Sakaguchi, A. Y. Loss of
`heterozygosity of chromosome 3p markers in small-cell
`lung cancer. Nature
`(Lond.), 329: 451-454, 1987.
`16. Kok, K., Osigna, J., Carri«, B., Davis, M. B., van der Hout, A. H., van der
`Veen, A. Y., Landsvater, R. M., de liji, L. F. M. H., Berendsen, H. H.,
`Postmus, P. E., Poppema, S., and Buys, C. H. M. Deletion of a DNA
`sequence at chromosomal
`region 3p21 in all major
`types of lung cancer.
`Nature (Lond.), 330: 578-581, 1987.
`17. Kovacs. G., and Kung, H. Nonhomologous chromatid exchange in hereditary
`and sporadic renal cell carcinomas. Proc. Nati. Acad. Sci. USA, 88: 194-
`198, 1991.
`18. Presti, J. C., Rao. P. H., Chen, Q.. Reuter, V. E., Li, F. P., Fair, W. R., and
`Jhanwar, S. C. Histopathological,
`cytogenetic, and molecular characteriza
`tion of renal cortical
`tumors. Cancer Res., 51: 1544-1552, 1991.
`19. Kinzler, K. W., Nilbert, M. C., Vogelstein, B., Bryan, T. M., Levy, D. B.,
`Smith, K. J.. Preisinger, A. C.. Hamilton, S. R., Hedge, P., Markhan, A.,
`Carlson, M., Joslyn, G., Groden,
`J., White, R., Miki, Y., Miyoshi, Y.,
`Nishisho,
`I., and Nakamura, Y. Identification of a gene located at chromo
`some 5q21 that
`is mutated in colorectal cancers. Science (Washington DC),
`251: 1366-1370. 1991.
`the Japanese Pathological Society, and
`20. Japanese Urologica! Association,
`Japan Radiological Society. General Rules for Clinical and Pathological
`Studies on Renal Cell Carcinoma, Ed. 1, pp. 68-73. Tokyo: Kanehara, 1983.
`
`5820
`
`West-Ward Exhibit 1099
`Morita 1991
`Page 004
`
`