* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download BRCA2 and p53 Mutations in Primary Breast
Survey
Document related concepts
Vectors in gene therapy wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Saethre–Chotzen syndrome wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
Polycomb Group Proteins and Cancer wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Microevolution wikipedia , lookup
Genome (book) wikipedia , lookup
Frameshift mutation wikipedia , lookup
BRCA mutation wikipedia , lookup
Point mutation wikipedia , lookup
Transcript
(CANCER RESEARCH 58. 859-862. March 1. 19981 Advances in Brief BRCA2 and p53 Mutations in Primary Breast Cancer in Relation to Genetic Instability1 Solveig Gretarsdottir, Steinunn Thorlacius, Rut Valgardsdottir, Sigfridur Gudlaugsdottir, Stefan Sigurdsson, Margret Steinarsdottir, Jon Gunnlaugur Jonasson, Kesara Anamthawat-Jonsson, and Jorunn E. Eyfjörd2 Molecular and Cell Biology Research Laboratory. Icelandic Cancer Society ¡S.Gr., S. T.. R. V.. S. Gu., S. S.. J. E. £./.• Cylogenetics Laboratory ¡M.S.] and Department of Pathology IM. S.. J. G. J.]. University Hospital of Iceland; and Biology Department. University of Iceland IK. A. J.j. Reykjavik. Iceland population.' Abstract This enabled us to analyze over 400 consecutive breast cancer cases for somatic p53 mutations, as well as for the germ-line BRCA2 founder mutation. A subset of samples were studied by conventional cytogenetic methods. FISH,4 and flow cytometry. thus The products of the BRCA breast cancer susceptibility genes have been implicated in cell cycle control and DNA repair. It has been suggested that mutations in the p53 gene are a necessary step in tumorigenesis in BRCA tumors. We tested samples from 402 breast cancer patients for germ-line allowing analysis of genetic instability in relation to these two genes. BRCA2 and p53 mutations in tumors. p53 mutations are more frequent in BRCA2 mutation carriers than they are in controls. Tumors with muta tions in either gene had multiple chromosomal abnormalities, as shown by cytogenetic analysis. Materialsand Methods The study material consisted of 402 breast tumor samples from the Icelandic Cancer Society Tumor Specimen Bank. Samples were screened for mutations in exons 5-8 of the p53 gene with the Introduction constant dénaturantgel electrophoresis method (19). p53 mutation status in relation to prognosis has been previously published for the majority of these samples (20-22). BRCA2 exon 9 fragments were PCR-amplified and run on Germ-line mutations in the BRCA I and BRCA2 genes have been shown to predispose to breast cancer (1-3), and the most common somatic changes found in breast tumors are mutations in the p53 gene (4). Both the BRCA and p53 genes have been implicated in cell cycle control (5-7). A number of studies have shown that p53 mutations are associated with genomic instability in tumors (8, 9), and recent studies on the functions of the BRCA proteins suggest that they are also involved in control of genomic integrity (10, 11). Both BRCA pro teins and p53 have been shown to interact with hRad51, a human homologue of the yeast Rad51 protein that is involved in recombina tion and DNA double-strand repair (10-12). BRCA1 and BRCA2 deficiencies cause cell lethality in early mouse embryos (13, 14), and hypersensitivity to -y-irradiation is seen in BRCA2 —¿/-cells (11). Recent studies show that BRCA1- and BRCA2-deficient embryos can 6% denaturing polyacrylamide gels for mutation detection (17). Mutants were identified by the presence of an extra alÃ-ele. Tumors were harvested directly for cytogenetic analyses or cultured on average for 6-7 days before harvesting (23). Analysis of karyotypes was done according to an International System for Human Cytogenetic Nomenclature (24). Karyotypic clones with one to three numerical changes (i.e.. additional or lacking chromosome) or a single structural change (i.e.. translocation or deletion) were listed as having simple clonal changes. Clones with more than three numerical changes and/or more than two structural changes were called complex. FISH was performed using PCR-amplified whole chromosome probes for chromosomes 1, 3, 16, and 17, as described by Anamthawat-Jonsson et al. (25), in which chromosomes 1 and 3 were labeled with red rhodamine and chromosomes 16 and 17 were labeled with green fluorescein. Flow cytometry was used to analyze the DNA content of the tumor cells. DNA ploidy index and S-phase assessment were performed as described previously (26). Histograms were classified as diploid (DNA index = 1), if there was a single G,/G, peak, or aneuploid (DNA index ^ 1), if there were at least two clearly distinct peaks, including multiploid, if more than two peaks were seen. The median S-phase value of <7% was defined as low S-phase fraction, and £7% was defined as high S-phase fraction. For statistical analysis, the jf test and Fisher's exact test were used as be partially rescued by p53 or p21 null mutations (13, 14). This suggests that cells with nonfunctional BRCA can only survive if they also have defective checkpoint control. Germ-line mutations in BRCA 1 or BRC A2 increase the risk of breast cancer, but even in these hereditary cases, breast cancer is a multistep process, and somatic mutations in other genes play a part in the tumor formation. Recent publications suggest that p53 abnormalities are a necessary step in tumorigenesis in BRCA1 carriers (15, 16). We wanted to examine this in an unselected group of breast cancer patients and test whether BRCA2 abnormalities were associated with genomic instability in tumors. Family and population studies indicate that there is only one mu tation in each BRCA gene in the Icelandic population, one rare BRCA1 mutation (D1692N) and a common BRCA2 founder mutation (999del5), found in 8% of all breast cancer cases in Iceland (17, 18). There are no indications of other BRCA 1 or BRCA2 mutations in the appropriate. The germ-line mutation screening was approved by the Icelandic Cancer Society Institutional Review Board. All DNA analyses were performed samples without personal identification. on Results p53 and BRCA2 Mutation Analysis. Samples from unselected breast cancer patients (n = 402) were screened for BRCA2 and p53 mutations. The BRCA2 999del5 germ-line mutation was detected in samples from 34 patients (8.4%), and somatic mutations in the p53 gene were detected in 72 (17.9%) tumors. The p53 mutations were distributed as follows: 25 mutations in exon 5 (35%), 9 in exon 6 (12%), 17 in exon 7 (24%), and 21 in exon 8 (29%). We compared the frequency and pattern of p53 mutations in tumors Received 11/4/97; accepted 1/19/98. The cosls 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. ' This work was supported by grants from The Icelandic Cancer Society Fund. Nordic Cancer Union. Icelandic Research Fund for Higher Education. University of Iceland Science Fund, and Icelandic Science Council. 2 To whom requests for reprints should be addressed, at Molecular and Cell Biology 1 Unpublished results. 4 The abbreviation used is: FISH, fluorescence in situ hybridization. Research Laboratory, Icelandic Cancer Society, P.O. Box 5420. 125 Reykjavik. Iceland. E-mail: [email protected]. 859 Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1998 American Association for Cancer Research. BRCA2 AND p53 MUTATIONS IN PRIMARY Table I Frequency- of p53 mutations in tumors from BKCA2 mutation carriers and noncarriersa p53wl BRCA2wt (n = 368) BRCA2mul (n = 34) 306 24 mulated (76.5%) and compared to the controls. There was, however, a clear difference observed in the complexity of the clonal changes. All of the clonal changes detected in the BRCA2 and p53 mutated tumors were complex as compared to about half of the clones in the mutant- p53mut 62 10 BRCA2wt. BREAST CARCINOMAS 0.09 BRCA2; negative tumors (Table 3). The chromosomal aberrations included both structural and numerical changes, they were not restricted to any particular chromosome and polyploidy was highly prevalent. How ever, among the primary breast tumor samples, chromosomes 1, 3, 16, Table 2 p5J mutations found in tumors from BRCA2 carriers and 17 were most frequently abnormal and were, therefore, selected Sample123456789IOExon5555677888Codon182173174141ND">NDNDNDfc298-302NDMutation1-base for FISH analysis with whole chromosome paint. The FISH analysis deletionGTG->ATGCAT—CTTTGC— confirmed the structural instability detected by G-banding and, fur thermore, showed that chromosomal rearrangements, gains, and losses CGCLarge were common. Multiple rearrangements of the painted chromosomes were detected in samples with BRCA2 germ-line mutations, p53 "p53wt, wild-type p53; p53mut. p53; wild-type BRCA2mul, mutated BRCA2. deletionLarge deletion14-bp somatic mutations, and both BRCA2 and p53 mutations (Fig. l, B, D, and F). Tumors with both BRCA2 and p53 mutations were not more complex than tumors with either mutation alone. S Phase. The proliferation rate of the tumors was examined. Both BRCA2 and p53 mutations were associated with a very high prolif- deletion " ND. not determined. * Samples from paraffin-embedded material. Mutation was not verified by sequencing. The mutations in samples 5, 8, and 10 were clearly detectable with the constant dénaturant gel electrophoresis method, which was used for the p53 mutation screening. This is a highly sensitive method that has enabled us to detect mutations in tissue with as little as 25% tumor cells. erative rate, with approximately 70% of tumors showing high mitotic activity, indicated by a high S-phase fraction (S7.00). This associa from BRCA2 carriers and noncarriers. We found that 10 BRCA2 mutation carriers (29%) had p53 mutations in their tumors or nearly twice as many as in the BRCA2 wild-type group (17%; Table 1). This Discussion Here, we screened samples from 402 unselected breast cancer patients for BRCA2 germ-line mutation and p53 mutations in primary difference was, however, not significant. The p53 mutation distribu tion in the BRCA2-positive group was similar to that found in the whole group (Table 2). Flow Cytometry, Cytogenetic, and FISH Analyses. tion was significant in all groups (Table 3). tumor samples. The frequency of BRCA2 mutation carriers (8.4%) was in agreement with our previous studies on Icelandic breast cancer patients (18). We detected mutations in exons 5-8 of the ¡>53 gene in Flow cyto- 17.9% of the tumors. This was also in agreement with our previous findings (20-22) and those of others (4, 27). metric analysis of DNA content was performed on 266 of the samples (Table 3). Aneuploidy was detected in 65% of all samples, and the prevalence was similar to that in BRCA2 mutants and wild-type samples. The same was true for the p53 mutated samples. Cytogenetic analysis was performed on fresh tumor samples from 60 patients (Table 3). There was no significant difference between samples from tumors with BRCA2 mutation compared to wild type, based on the occurrence of simple and/or complex clonal changes. The Cytogenetic and flow cytometry data combined showed that 73% of the samples from BRCA2 carriers were aneuploid, compared to 63% of the BRCA2/p53 normal samples. The instability trend was further strengthened if BRCA2 and p53 mutated samples were pooled p53 mutations were found to be more common in tumors from BRCA2 mutation carriers than they were in controls. However, the frequency was not as high as those recently reported in breast tumors from BRCA1 mutation carriers (15). In that study, p53 mutations were found in eight of eight tumors with known BRCA1 mutations, and the majority of p53 mutations were located in exon 5. The distribution of p53 mutations found in our study was the same in BRCA2 mutation carriers as in the whole group. None of the mutations reported in the BRCA1 carriers by Crook et al. (15) were found. p53 protein overexpression in BRCA1 tumors from high-risk families also indicate a fairly high frequency of Table 3 Association of BRCA2 and/or p53 mutated samples with flow cytometry and Cytogenetic data The mutated samples were compared to the wild type using the Fisher's exact test. Abbreviations are as in Table 1. BRCA2mutPloidy P450.5 p53mul p53mut6217(27.5)45 or BRCA2wl/p53wt2040.17 266)TotalDiploid (n = (%)Aneuploid (%)S-phase 255)Total<7.00(%)27.00 (n = (78)420.04 0.0635 10(22) (%)Cytogenetics 60)TotalNo (n = (69)110.7 <%)Clonal clonal changes (%)Clonal changes 32)TotalSimple changes (n = 0.57(64)70.12 (%)Complex (%)Ploidy 217)TotalNormal and cytogenetics (n = 0.067(100)470.390 77(38)127(62)1980.0009 (72.5)5717(30)40 0.00629 13(31) 109(55)89 (70)177(41)10(59)10010(100)6415(23)49 (45)430.8 4 (36) (49)22(51)220.013 21 10(45)12 (55)2130.05 (%)Abnormal 0.01738 9(20) (%)268(31)18(69)227(32)15(68)83(37)5(63)505(100)267(27)19(73)P(80)BRCA2mut (37)134(63) 79 (77)P 860 Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1998 American Association for Cancer Research. BRCA2 AND p53 MUTATIONS IN PRIMARY BREAST CARCINOMAS i *tt •¿â€¢J Fig. 1. FISH with whole chromosome painting showing structural abnormalities in metaphases (ß.O, and F). The cells were counterslained with 4'.6-diamidino-2-phenylindo!e (A, C. and £7). &•«/(' bar. K) ¿im.The chromosome abnormalities involved both structural and numerical changes, and they are of very complex type, based on G-handing and FISH analysis. A and R, near-triploid cell from a tumor from a BRCA2 mutant carrier after painting of chromosomes I and 16 (red and gMtii/btOtttetOCtt respectively). Chromosome I is involved in eight translocations and shows overall gain of material, but chromosome 16 is seen in one translocatton and shows loss of material. C and D, near-triploid cell from p53 mutated tumor after painting of chromosomes 1 and 16 (red and green fluorescence, respectively). Here, chromosome 1 lakes part in two trän s locations, and chromosome 16 shows gain. E and F. near-diploid cell from a BRCA2 and p53 mutated tumor after painting of chromosomes 3 and 17 (red and green fluorescence, respectively). In this cell, four rearrangements can be seen, one of which is a translocation between the two painted chromosomes. 861 Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1998 American Association for Cancer Research. BRCA2 AND p53 MUTATIONS [N PRIMARY BREAST CARCINOMAS mutations p53 abnormalities, 40 and 70%, respectively (16,27). p53 overexpression was not associated with BRCA2 mutation in our study (data not shown). The samples analyzed in this study were from breast cancer patients unselected for family history. Ten of the BRCA2 mutation carriers were, however, found to belong to known high-risk families (17). Of those, in human tumors and cell lines. Nucleic Acids Res.. 22: 3551-3555, 1994. Vaughn, J. P., Cirisano, F. D., Huper. G., Berchuck, A., Futreal, P. A., Marks, J. R., and Iglehart, J. D. Cell cycle control of BRCA2. Cancer Res., 56: 4590-4594, 1996. Rajan, J. V., Marquis, S. T., Gardner, H. P.. Chodosh, L. A., Rajan, J. V., Wang, M., Marquis. S. T.. and Chodosh, L. A. Developmental expression of Brca2 colocalizes with Brea I and is associated with proliferation and differentiation in multiple tissues. Dev. Biol.. 184: 385-401, 1997. Levine. A. J. p53, the cellular gatekeeper for growth and division. Cell, 88: 323-331, only three had p53 mutations, and there was one additional case of p53 overexpression. Soboi et al. (16) suggest that specific mutations in the BRCAI gene are more often associated with p53 abnormalities as meas ured by abnormal staining. Because all of the BRCA2 mutation carriers in our study have the same 999del5 mutation, no conclusion can be drawn about association between p53 abnormalities and BRCA2 mutations in general. As mentioned previously, only one rare BRCAI mutation has been found in this population. A single case of this mutation was found in this cohort. Neither p53 mutation nor abnormal p53 staining was detected in a tumor sample from this patient. We previously described a significant association between p53 abnor malities and genomic instability in primary breast tumors (9, 22). This study supports this and, furthermore, shows that BRCA2 mutated tumors have complex chromosomal changes as well. Culturing primary breast tumor cells is difficult, and in particular, it is hard to get cells with complex karyotypes to divide. The most abnormal cancer cells may, therefore, be lost in the process of cell culture and chromosome harvest ing. The BRCA2 samples were, indeed, highly complex showing multi ple chromosomal rearrangements (e.g.. Fig. Iß),which seems to fit the notion of BRCA2 involvement in double-strand DNA repair. It is well known that p53 participates in cell cycle control by activating p21 in response to DNA damage (29). It has been shown that the expression of the BRCA genes is cell cycle dependent (5, 30), and in a recent study, BRCAI was found to activate p21 in a p53-independent manner (31 ). It is as yet unknown how BRCA2 participates in cell cycle control. We found that tumors from BRCA2 mutation carriers were highly proliferative, as judged by high S phase. This was also true for tumors without additional p53 abnormalities, suggesting a possible role for BRCA2 in cell cycle control. In conclusion, our results support findings that implicate BRCA2 in DNA repair and cell cycle control. Inactivation of p53 may be important in initiation of tumorigenesis in BRCA2 carriers. However, our results show that two of three of the BRCA2 mutated tumors have normal p53 and, therefore, inactivation of other integrity control genes must be involved in these tumors, allowing cells with highly abnormal chromosomes to go through the cell cycle. 10. 12. 14. 16. 17 18 19 21 22 Acknowledgments 23 We thank the Department of Pathology. National Hospital of Iceland, for providing samples. We thank the staff at the Department of Pathology for immunostaining. the staff at Cancer Registry for assistance with population -,5 data, and the Icelandic Agricultural Institute for FISH facilities. We also thank Dr. Helga Ögmundsdottir for valuable discussion and comments. 26 References 1. Wooster. R.. Bignell. G., Lancaster, J., Swift, S., Seal, S., Mangion, J., Collins, N., Gregory, S.. Gumbs, C., and Micklem, G. Identification of the breast cancer suscep tibility gene BKCA2. Nature (Lond.l, 378: 789-792. 1995. 2. Miki, Y., Katagiri. T.. Kasumi. F., Yoshimoto. T.. and Nakamura, Y. Mutation analysis in the BRCA2 gene in primary breast cancers. Nat. Genet.. 13: 245-247. 19%. 3. Tavtigian. S. V., Simard, J., Rommens. J.. Couch. F., Shattuck-Eidens, D.. Neuhausen. S.. Merajver. S.. Thorlacius. S.. Off«.K.. Stoppa-Lyonnet. D.. Belanger. C.. Bell. R.. Berry, S., Bogden, R.. Chen, Q., Davis, T.. Dumont, M., Frye. C.. Hattier, T., Jammulapati. S.. Janecki, T.. Jiang, P., Kehrer, R.. Leblanc. J. F.. Mitchell, J. T., McAnhur-Morrison. J., Nguyen, K.. Peng, Y., Samson, C.. Schroeder. M., Snyder, S. C.. Steele. L., Stringfellow, M., Slroup, C.. Swedlund. B., Swensen, J., Teng, D., Thomas. A., Tran, T., Tran, T., Tranchant. M.. Weaver-Feldhaus, J., Wong, A. K. C.. Shizuya, H., Eyfjörd.J. E.. Cannon-Albright. L.. Labrie, F.. Skolnick. M.. Weber. B., Kamb. A., and Goldgar. D. E. The complete BRCA2 gèneand mutations in chromosome 13q-linked kindreds. Nat. Genet.. 12: 333-337. 1996. 4. Hollstein, M.. Rice, K.. Greenblatt. M. S., Soussi. T.. Fuchs, R.. Sorlie, T., Hovig, E., Smilh-Sorensen, B., Montesano, R., and Harris, C. C. Database of p5J gene somatic 27 ° 1997. Livingstone, L. R., White, A.. Sprouse, J., Livanos, E., Jacks, T., and Tlsty, T. D. Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell. 70: 923-935, 1992. Eyfjord, J. E., Thorlacius, S., Steinarsdottir. M.. Valgardsdoltir. R.. Ögmundsdottir. H., and Anamthawat-Jonsson, K. p53 abnormalities and genomic instability in pri mary human breast carcinomas. Cancer Res., 55: 645-651, 1995. Scully, R., Chen, J., Plug, A., Xiao, Y., Weaver, D., Feunteun, J., Ashley, T., and Livingston. D. M. Association of BRCAI with Rad51 in mitotic and meiotic cells. Cell, 88: 265-275, 1997. Sharan. S. K.. Morimalsu. M., Albrecht, U.. Lim. D. S.. Regel, E., Dinh. C.. Sands. A.. Eichele, G., Hasty. P., and Bradley, A. Embryonic lethality and radiation hypersensitivity mediated by RadSI in mice lacking Brca2. Nature (Lond.). 386: 804-810, 1997. Buchhop. S.. Gibson, M. K., Wang, X. W., Wagner. P.. Sturzbecher. H. W., and Harris. C. C. Interaction of p53 with the human rad51 protein. Nucleic Acids Res., 25: 3868-3874, 1997. Hakem. R., de la Pompa, J. L.. Elia, A., Potter, J., and Mak, T. W. Partial rescue of Brcal early embryonic lethality by p53 or p2l null mutation. Nat. Genet., 16: 298-302. 1997. Ludwig. T.. Chapman. D. L.. Papaioannou. V. E., and Efstratiadis. A. Targeted mutations of breast cancer susceptibility gene homologs in mice: lethal phenotypes of Brcal, Brca2. Brcal/Brca2, Brcal/p53. and Brca2/p53 nullizygous embryos. Genes Dev., //: 1226-1241, 1997. Crook. T.. Crossland. S.. Cromplon. M. R.. Osin. P.. and Gusterson, B. A. p53 mutations in Brea I-associated familial breast cancer. Lancet, 350: 638-639. 1997. Sobol, H.. Stoppa-Lyonnet, D., Bressac-De Paillerets, B., Peyrat, J. P., Guinebretiere, J-M.. Jacquemier. J.. Eisinger. F.. and Birnbaum. D. BRCAl-p53 relationship in hereditary breast cancer. Int. J. Oncol., 10: 349-353, 1997. Thorlacius. S.. Olafsdottir. G.. Tryggvadottir, L.. Neuhausen, S.. Jonasson, J. G., Tavtigian, S. V., Tulinius. H.. Ögmundsdottir. H. M., and Eyfjord, J. E. A single BRCA2 mutation in male and female breast cancer families from Iceland with varied cancer phenotypes. Nat. Genet.. 13: 117-119, 1996. Thorlacius, S.. Sigurdsson, S.. Bjamadotlir. H.. Olafsdottir. G.. Jonasson. J. G.. Tryggvadottir. L., Tulinius. H., and Eyfjord, J. E. Study of a single BRCA2 mutation with high carrier frequency in a small population. Am. J. Hum. Genet., 60: 1079-1084, 1997. Borresen, A. L., Hovig. E., Smilh-Sorensen, B., Malkin, D.. Lystad, S.. Andersen, T. I.. Nesland, J. M.. Isselbacher. K. J., and Friend, S. H. Constant dénaturantgel electrophoresis as a rapid screening technique for p53 mutations. Proc. Nail. Acad. Sci. USA. 88: 8405-8409, 1991. Thorlacius, S.. Thorgilsson. B.. Bjömsson, J.. Tryggvadottir, L., Börresen, A-L., Ögmundsdottir, H. M.. and Eyfjörd,J. E. TP53 mutaûonsand abnormal p53 protein staining in breast carcinomas related to prognosis. Eur. J. Cancer, 31A: 1856-1861. 1995. Gretarsdottir, S., Tryggvadottir, L.. Jonasson, J. G., Sigurdsson, H., Olafsdottir, K.. Agnarsson, B. A.. Ögmundsdottir.H. M., and Eyfjörd,J. E. TP53 mutation analyses on breast carcinomas; a study of paraffin embedded archival material. Br. J. Cancer, 74: 555-561. 19%. Valgardsdoltir. R.. Tryggvadottir, L., Steinarsdottir. M.. Olafsdottir, K.. Jonasdoltir, S.. Jonasson, J. G., Ögmundsdottir. H., and Eyfjörd, J. E. Genomic instability and poor prognosis associated with abnormal p53 in breast carcinomas. Molecular and immunohistochemical analysis. APMIS, 105: 121-130. 1997. Steinarsdottir, M.. Petursdottir, I.. Snorradottìr, S.. Eyfjord, J. E., and Ögmundsdottir.H. M. Cytogenetic studies of breast carcinomas: different karyotypic profiles delected by direct harvesting and short-term culture. Genes Chromosomes Cancer. 13: 239-248, 1995. Mitelman, F. 1SCN Guidelines for Cancer Cytogenetics: Supplement to an International System for Human Cytogenetic Nomenclature. Basel: S. Karger. 1995. Anamthawat-Jonsson. K., Eyfjord. J. E.. Ögmundsdottir. H. M.. Petursdottir. I., and Steinarsdottir, M. Instability of chromosomes 1. 3, 16, and 17 in primary breast carcinomas inferred by fluorescence in silu hybridization. Cancer Genet. Cytogenet., 88: 1-7, 1996. Jonasson, J. G.. and Hrafnkelsson, J. Nuclear DNA analysis and prognosis in carcinoma of the thyroid gland. A nationwide study in Iceland on carcinomas diagnosed 1955-1990. Virchows Arch. A. 425: 349-355, 1994. Bergh. J.. Norberg, T., Sjögren. S., Lindgren, A., and Holmberg. L. Complete sequencing of thep5.? gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy. Nat. Med., I: 1029-1034, 1995. EI-Deiry. W. S.. Harper, J. W.. O'Connor. P. M.. Velculescu, V. E., Canman. C. E., Jackman. J., Pietenpol, J., Burrell. M.. Hill, D. E., Wang, Y.. Wiman, K. G., Mercer. W. E., Kastan, M. B., Kohn, K. W., Elledge, S. J., Kinzler, K. W., and Vogelslein, B. WAF1/CIP1 is induced in p53-mediated G, arrest and apoptosis. Cancer Res., 54: 1169-1174, 1994. 29 Rajan. J. V., Wang, M.. Marquis, S. T.. and Chodosh. L. A. Brca2 is coordinately regulated with Brcal during proliferation and differentiation in mammary epithelial cells. Proc. Nati. Acad. Sci. USA. 93: 13078-13083, 1996. ^0 Somasundaram. K.. Zhang. H.. Zeng. Y. X., Houvras, Y., Peng, Y., Zhang. H.. Wu. G. S., Licht, J. D.. Weber. B. L.. and El-Deiry, W. S. Arrest of the cell cycle by the tumor-suppressor BRCAI requires the CDK-inhibitor p21WAFl/CiPl. Nature (Lond.). .«9: 187-190, 1997. 862 Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1998 American Association for Cancer Research. BRCA2 and p53 Mutations in Primary Breast Cancer in Relation to Genetic Instability Solveig Gretarsdottir, Steinunn Thorlacius, Rut Valgardsdottir, et al. Cancer Res 1998;58:859-862. Updated version E-mail alerts Reprints and Subscriptions Permissions Access the most recent version of this article at: http://cancerres.aacrjournals.org/content/58/5/859 Sign up to receive free email-alerts related to this article or journal. To order reprints of this article or to subscribe to the journal, contact the AACR Publications Department at [email protected]. To request permission to re-use all or part of this article, contact the AACR Publications Department at [email protected]. Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1998 American Association for Cancer Research.