MYC High Level Gene Amplification Is a Distinctive Feature of Angiosarcomas after Irradiation or Chronic Lymphedema
2009; Elsevier BV; Volume: 176; Issue: 1 Linguagem: Inglês
10.2353/ajpath.2010.090637
ISSN1525-2191
AutoresJohanna Manner, Bernhard Radlwimmer, Peter Hohenberger, Katharina Mößinger, Stefan Küffer, Christian Sauer, Djeda Belharazem, Andreas Zettl, Jean‐Michel Coindre, Christian Hallermann, Jörg T. Hartmann, D. Katenkamp, Kathrin Katenkamp, Patrick Schöffski, Raf Sciot, Agnieszka Woźniak, Peter Lichter, Alexander Marx, Philipp Ströbel,
Tópico(s)Sarcoma Diagnosis and Treatment
ResumoAngiosarcomas (AS) are rare vascular malignancies that arise either de novo as primary tumors or secondary to irradiation or chronic lymphedema. The cytogenetics of angiosarcomas are poorly characterized. We applied array-comparative genomic hybridization as a screening method to identify recurrent alterations in 22 cases. Recurrent genetic alterations were identified only in secondary but not in primary AS. The most frequent recurrent alterations were high level amplifications on chromosome 8q24.21 (50%), followed by 10p12.33 (33%) and 5q35.3 (11%). Fluorescence in situ hybridization analysis in 28 primary and 33 secondary angiosarcomas (31 tumors secondary to irradiation, 2 tumors secondary to chronic lymphedema) confirmed high level amplification of MYC on chromosome 8q24.21 as a recurrent genetic alteration found exclusively in 55% of AS secondary to irradiation or chronic lymphedema, but not in primary AS. Amplification of MYC did not predispose to high grade morphology or increased cell turnover. In conclusion, despite their identical morphology, secondary AS are genetically different from primary AS and are characterized by a high frequency of high level amplifications of MYC. This finding may have implications both for the diagnosis and treatment of these tumors. Angiosarcomas (AS) are rare vascular malignancies that arise either de novo as primary tumors or secondary to irradiation or chronic lymphedema. The cytogenetics of angiosarcomas are poorly characterized. We applied array-comparative genomic hybridization as a screening method to identify recurrent alterations in 22 cases. Recurrent genetic alterations were identified only in secondary but not in primary AS. The most frequent recurrent alterations were high level amplifications on chromosome 8q24.21 (50%), followed by 10p12.33 (33%) and 5q35.3 (11%). Fluorescence in situ hybridization analysis in 28 primary and 33 secondary angiosarcomas (31 tumors secondary to irradiation, 2 tumors secondary to chronic lymphedema) confirmed high level amplification of MYC on chromosome 8q24.21 as a recurrent genetic alteration found exclusively in 55% of AS secondary to irradiation or chronic lymphedema, but not in primary AS. Amplification of MYC did not predispose to high grade morphology or increased cell turnover. In conclusion, despite their identical morphology, secondary AS are genetically different from primary AS and are characterized by a high frequency of high level amplifications of MYC. This finding may have implications both for the diagnosis and treatment of these tumors. Angiosarcomas (AS) are rare sarcomas with morphological and functional properties of endothelial cells.1Weiss SW Goldblum JR Malignant vascular tumors.in: Weiss SW Goldblum JR Soft tissue tumors. Mosby Elsevier, 2008: 703-750Google Scholar AS represent 10 years.11Hodgson NC Bowen-Wells C Moffat F Franceschi D Avisar E Angiosarcomas of the breast: a review of 70 cases.Am J Clin Oncol. 2007; 30: 570-573Crossref PubMed Scopus (92) Google Scholar, 12Vorburger SA Xing Y Hunt KK Lakin GE Benjamin RS Feig BW Pisters PW Ballo MT Chen L Trent 3rd, J Burgess M Patel S Pollock RE Cormier JN Angiosarcoma of the breast.Cancer. 2005; 104: 2682-2688Crossref PubMed Scopus (160) Google Scholar, 13Adhikari D Hajdu SI Levine D Post-radiation angiosarcoma and bilateral mastectomy.Ann Clin Lab Sci. 2002; 32: 428-433PubMed Google Scholar, 14Monroe AT Feigenberg SJ Mendenhall NP Angiosarcoma after breast-conserving therapy.Cancer. 2003; 97: 1832-1840Crossref PubMed Scopus (170) Google Scholar Relatively little is known about the genetic changes in postradiation sarcomas in general15Tarkkanen M Wiklund TA Virolainen MJ Larramendy ML Mandahl N Mertens F Blomqvist CP Tukiainen EJ Miettinen MM Elomaa AI Knuutila YS Comparative genomic hybridization of postirradiation sarcomas.Cancer. 2001; 92: 1992-1998Crossref PubMed Scopus (28) Google Scholar, 16Mertens F Larramendy M Gustavsson A Gisselsson D Rydholm A Brosjo O Mitelman F Knuutila S Mandahl N Radiation-associated sarcomas are characterized by complex karyotypes with frequent rearrangements of chromosome arm 3p.Cancer Genet Cytogenet. 2000; 116: 89-96Abstract Full Text Full Text PDF PubMed Scopus (51) Google Scholar, 17Nakanishi H Tomita Y Myoui A Yoshikawa H Sakai K Kato Y Ochi T Aozasa K Mutation of the p53 gene in postradiation sarcoma.Lab Invest. 1998; 78: 727-733PubMed Google Scholar, 18Brachman DG Hallahan DE Beckett MA Yandell DW Weichselbaum RR p53 gene mutations and abnormal retinoblastoma protein in radiation-induced human sarcomas.Cancer Res. 1991; 51: 6393-6396PubMed Google Scholar and in sAS in particular, where published genetic studies are limited to case reports or small case collections.19Gil-Benso R Lopez-Gines C Soriano P Almenar S Vazquez C Llombart-Bosch A Cytogenetic study of angiosarcoma of the breast.Genes Chromosomes Cancer. 1994; 10: 210-212Crossref PubMed Scopus (24) Google Scholar, 20Naka N Tomita Y Nakanishi H Araki N Hongyo T Ochi T Aozasa K Mutations of p53 tumor-suppressor gene in angiosarcoma.Int J Cancer. 1997; 71: 952-955Crossref PubMed Scopus (79) Google Scholar, 21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, 22Cerilli LA Huffman HT Anand A Primary renal angiosarcoma: a case report with immunohistochemical, ultrastructural, and cytogenetic features and review of the literature.Arch Pathol Lab Med. 1998; 122: 929-935PubMed Google Scholar, 23Schuborg C Mertens F Rydholm A Brosjo O Dictor M Mitelman F Mandahl N Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue.Cancer Genet Cytogenet. 1998; 100: 52-56Abstract Full Text PDF PubMed Scopus (30) Google Scholar, 24Wong KF So CC Wong N Siu LL Kwong YL Chan JK Sinonasal angiosarcoma with marrow involvement at presentation mimicking malignant lymphoma: cytogenetic analysis using multiple techniques.Cancer Genet Cytogenet. 2001; 129: 64-68Abstract Full Text Full Text PDF PubMed Scopus (27) Google Scholar, 25Zu Y Perle MA Yan Z Liu J Kumar A Waisman J Chromosomal abnormalities and p53 gene mutation in a cardiac angiosarcoma.Appl Immunohistochem Mol Morphol. 2001; 9: 24-28Crossref PubMed Scopus (32) Google Scholar, 26Mandahl N Jin YS Heim S Willen H Wennerberg J Biorklund A Mitelman F Trisomy 5 and loss of the Y chromosome as the sole cytogenetic anomalies in a cavernous hemangioma/angiosarcoma.Genes Chromosomes Cancer. 1990; 1: 315-316Crossref PubMed Scopus (28) Google Scholar, 27Kindblom LG Stenman G Angervall L Morphological and cytogenetic studies of angiosarcoma in Stewart-Treves syndrome.Virchows Arch A Pathol Anat Histopathol. 1991; 419: 439-445Crossref PubMed Scopus (39) Google Scholar, 28Van den Berg E Van Oven MW de Jong B Dam A Wiersema J Dijkhuizen T Hoekstra HJ Molenaar WM Comparison of cytogenetic abnormalities and deoxyribonucleic acid ploidy of benign, borderline malignant, and different grades of malignant soft tissue tumors.Lab Invest. 1994; 70: 307-313PubMed Google Scholar Probably due to the small case numbers, available data have so far failed to show consistent recurrent chromosomal abnormalities. In this report, we studied clinicopathologic and molecular genetic features in 61 primary and secondary AS from multiple international institutions. The clinicopathologic data of studied cases are shown in Table 1. Paraffin blocks of histologically proven angiosarcomas were retrospectively collected from the collaborating institutions in Germany, France and Belgium. All tumors were stratified by histological grade according to the scoring system of the Fédération Nationale des Centres de Lutte Contre le Cancer29Guillou L Coindre JM Bonichon F Nguyen BB Terrier P Collin F Vilain MO Mandard AM Le Doussal V Leroux A Jacquemier J Duplay H Sastre-Garau X Costa J Comparative study of the National Cancer Institute and French Federation of Cancer Centers Sarcoma Group grading systems in a population of 410 adult patients with soft tissue sarcoma.J Clin Oncol. 1997; 15: 350-362Crossref PubMed Scopus (693) Google Scholar by two of the authors (A.M. and Ph.S.).Table 1Summary of Clinicopathological Findings in Primary and Secondary AngiosarcomaspAS (n = 28)sAS (n = 33)n =%n =%Male1554515Female13462885Tumor site Breast272061 Skin8297*Including 2 cases associated with chronic lymphedema.21 Soft tissue31126 Bone31113 Heart41400 Spleen31100 Liver1400 Other414310FNCLCC grade 19321030 21139824 38361539pAS, primary angiosarcomas; sAS, secondary angiosarcomas.* Including 2 cases associated with chronic lymphedema. Open table in a new tab pAS, primary angiosarcomas; sAS, secondary angiosarcomas. Immunohistochemistry was performed according to standard protocols.30Yoneda S Marx A Muller-Hermelink HK Low-grade metaplastic carcinomas of the thymus: biphasic thymic epithelial tumors with mesenchymal metaplasia–an update.Pathol Res Pract. 1999; 195: 555-563Crossref PubMed Scopus (19) Google Scholar Primary antibodies used were CD34 (1:500; Beckman Coulter, Krefeld, Germany) and CD31 (1:500, Dako, Hamburg, Germany) and ki67 (1:800, Dako). Proliferation was quantified under a light microscope by counting ki-67 stained nuclei per 100 tumor cells in randomly chosen high power fields. Apoptosis was measured using the commercially available terminal deoxynucleotidyl transferase dUTP nick-end labeling kit (In Situ Cell Death Detection Kit, Roche Applied Science, Mannheim, Germany), according to the manufacturer's instructions and quantitated under a Zeiss Axiophot fluorescence microscope by counting fluorescein isothiocyanate-stained nuclei per 100 tumor cell nuclei counterstained with 4′,6-diamidin-2′-phenylindol-dihydrochloride (Linaris, Wertheim, Germany). Array-comparative genomic hybridization was performed as previously described,31Zielinski B Gratias S Toedt G Mendrzyk F Stange DE Radlwimmer B Lohmann DR Lichter P Detection of chromosomal imbalances in retinoblastoma by matrix-based comparative genomic hybridization.Genes Chromosomes Cancer. 2005; 43: 294-301Crossref PubMed Scopus (97) Google Scholar, 32Hoischen A Landwehr C Kabisch S Ding XQ Trost D Stropahl G Wigger M Radlwimmer B Weber RG Haffner D Array-CGH in unclear syndromic nephropathies identifies a microdeletion in Xq22.3-q23.Pediatr Nephrol. 2009; PubMed Google Scholar, 33Fensterer H Radlwimmer B Strater J Buchholz M Aust DE Julie C Radvanyi F Nordlinger B Belluco C Van Cutsem E Kohne CH Kestler HA Schwaenen C Nessling M Lutz MP Lichter P Gress TM Matrix-comparative genomic hybridization from multicenter formalin-fixed paraffin-embedded colorectal cancer tissue blocks.BMC Cancer. 2007; 7: 58Crossref PubMed Scopus (19) Google Scholar using genomic DNA isolated from fresh frozen tumor material on microarrays with more than 8000 large insert clones (8-k array). The array contained the Sanger Center 1-Mb clone set covering the genome at an average resolution of approximately 1Mb, 3000 gene- and region-specific RCPI (Roswell Park Cancer Institute database) (RZPD, Berlin, Germany) and CalTech (Invitrogen, Carlsbad, CA) bacterial artificial chromosome clones. Arrays were hybridized with Cy3-labeled test DNA and Cy5-labeled reference DNA. Reference DNA pools were generated from ten healthy women and from ten healthy men. Array-comparative genomic hybridization data were processed using the ChipYard framework (Heidelberg, Germany). Diagnostic thresholds were defined by extensive positive and negative control experiments as ±7 SD of the mean. Fluorescence in situ hybridization was performed as previously described.34Hildenbrand R Niedergethmann M Marx A Belharazem D Allgayer H Schleger C Strobel P Amplification of the urokinase-type-plasminogen activator-receptor (uPAR) gene in ductal pancreatic carcinomas identifies a clinical high risk group.Am J Pathol. 2009; 174: 2246-2253Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar Hybridization was performed using a P1 probe for MYC labeled with digoxigenin by nick translation (RMC08P001; provided by the Cancer Center, University of San Francisco, San Francisco, CA) and an α-satellite probe for the centromeric region of chromosome 8 (Vysis Inc., Downers Grove, IL). For detection of the MYC probe, the tyramide signal amplification system from NEN Life Science GmbH (Köln, Germany) was applied according to the manufacturer's recommendations. In brief, after a 30-minute blocking step, slides were treated with peroxidase-conjugated anti-digoxigenin antibody (1:50 in tyramide signal amplification blocking solution; Roche Diagnostics GmbH) for 90 minutes at 37°C. For visualization of the probe, slides were incubated for 7 minutes with tyramide-Cy3 (1:50 in tyramide signal amplification solution). Interphase nuclei were counterstained with 0.5 μg/μL of 4,6-diamidino-2-phenylindole in Vectashield mounting medium (Vector Laboratories, Burlingame, CA). At least 200 nuclei per sample were assessed and scored semiquantitatively. Amplification was assumed if more than two times as many gene signals compared with centromere signals were detected in >25% of tumor cells. Detection of >9 gene signals or tight clusters of at least 10 gene signals was considered high level amplification. Statistical differences between categoric variables were determined using Yates corrected Chi-Square test. Comparisons between groups were analyzed using Mann-Whitney-U-test, as provided by the SPSS software (Version 2000, SPSS Inc., Chicago, IL). For all analyses, a P value <0.05 was considered significant, unless otherwise stated. Cases were retrospectively collected in reference sarcoma centers. A summary of the clinicopathologic data are shown in Table 1. pAS and sAS occurred in the same age groups (pAS: age median 69 years [range, 32 to 85]; sAS: age median 73 years [range, 34 to 88]). The majority (61%) of sAS were observed in the breast of females with a history of irradiation for mammary carcinomas, followed by skin (21%) and soft tissues (6%); two cases arose in the setting of chronic lymphedema. There were no AS associated with a history of thorotrast exposure in this series. In contrast to the marked female preponderance among sAS, males and females were affected with equal frequency in the pAS group. Skin tumors made up roughly one third of the cases, the remaining cases were located in soft tissues and internal organs. The relative distribution of histological tumor grades 1 to 3 was identical among pAS and sAS. Tumors with epitheloid features were slightly more frequent in the pAS (six cases) than in the sAS group (two cases). Twenty-two cases (8 pAS; 14 sAS) were analyzed by array-comparative genomic hybridization. Of these, 10 sAS (nine irradiated cases and one case associated with chronic lymphedema) showed a total of 16 recurrent alterations (Table 2 + 3, Figure 1; full data available online under in Supplemental Table S1 at http://ajp.amjpathol.org), while no recurrent alterations were observed in pAS. The most frequent alterations in sAS were high level amplifications on chromosome 8q24.21 (eight cases), followed by amplifications on chromosome 10p12.33 (six cases) and amplifications on chromosome 5q35.3 (two cases). Amplifications on chromosome 8q24.21 were frequently observed in combination with other amplifications, most often on chromosome 10p12.33 (six cases), but also chromosome 5q35.3 (two cases).Table 2Chromosome Loci Displaying Recurrent Copy-Number Amplifications or Homozygous DeletionsMappingStart [Mbp]End [Mbp]Size [Mbp]TypepAS (n = 8)sAS (n = 14)5q35.3178.69180.151.46Amp028q24.21128.04129.041.00Amp0810p12.3317.8418.941.10Amp06pAS, primary angiosarcomas; sAS, secondary angiosarcomas; Del, deletion; Amp, amplification. Open table in a new tab Table 3Clinicopathologic Characteristics of Angiosarcomas with Chromosomal AlterationsCase no.EtiologyLocalizationGradeDetected chromosomal aberrationsHA 96RadiationBreast3Amp 5q35.3; amp 8q24.21BO 5315RadiationBreast2Amp 5q35.3; amp 8q24.21; amp 10p12.33; amp 19q13.32BO 3108RadiationBreast3Amp 8q24.21; amp 10p12.33K 6911RadiationBreast1Amp 8q24.21; amp 10p12.33K 7334RadiationBreast2Amp 8q24.21; amp 10p12.33BO 4592RadiationBreast1Amp 8q24.21K 12803RadiationSoft tissue2Amp 8q24.21GN 516RadiationBreast2Amp 10p12.33EF 73Chronic LESoft tissue3Amp 8q24.21; amp 10p12.33Chronic LE, chronic lymphedema. Open table in a new tab pAS, primary angiosarcomas; sAS, secondary angiosarcomas; Del, deletion; Amp, amplification. Chronic LE, chronic lymphedema. Since the most likely candidate gene for high level amplification in the chromosomal region 8q24.21 was MYC, we next applied fluorescence in situ hybridization to verify this assumption (Figure 2, A–C). In an analysis of 61 cases (28 pAS, 33 sAS), a total of 18 cases with high level gene amplifications of MYC were observed. Of these, all 18 (100%) were sAS (P < 0.001) (Table 4). Of note, apart from 16 radiation-induced AS, also both of the AS associated with chronic lymphedema showed MYC amplifications. By morphology, sAS with MYC high level amplifications were not different from either sAS or pAS without MYC amplification. Cases with high histological grade (G3) were observed with the same frequency in both groups (P > 0.05) (Table 4). Since MYC plays an important role in cell cycle regulation, we also analyzed proliferation and apoptosis by immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, respectively, but found no differences between AS with and without MYC gene amplifications (P > 0.05) (Table 4).Table 4Statistical Comparison of Primary and Secondary AngiosarcomasMYC high levelMYC nc.pAS0 (0%)28 (100%)28sAS18 (55%)15 (45%)33184361P < 0.0001MYC high levelMYC nc.G35 (45%)6 (55%)11Other11 (55%)9 (45%)20161531P > 0.05MYC high levelMYC nc.Ki67 (%)45 ± 18.039 ± 13.4P > 0.05TUNEL (%)4 ± 4.07 ± 10.4P > 0.05pAS, primary angiosarcomas; sAS, secondary angiosarcomas; nc, no change.High frequency of MYC high level gene amplifications among sAS, but not pAS (Chi-Square test). High tumor grade was as frequent in sAS with MYC high level gene amplifications as in sAS without (Chi-Square test). Proliferation (ki67-positive cells per 100 tumor cells) and apoptosis rates (terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive cells per 100 tumor cells) were identical among pAS and sAS (Mann-Whitney-U test). Open table in a new tab pAS, primary angiosarcomas; sAS, secondary angiosarcomas; nc, no change. High frequency of MYC high level gene amplifications among sAS, but not pAS (Chi-Square test). High tumor grade was as frequent in sAS with MYC high level gene amplifications as in sAS without (Chi-Square test). Proliferation (ki67-positive cells per 100 tumor cells) and apoptosis rates (terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive cells per 100 tumor cells) were identical among pAS and sAS (Mann-Whitney-U test). In this comprehensive analysis, we show for the first time that primary and secondary AS are genetically different entities, although these tumors are undistinguishable by morphology. Irradiation and chronic lymphedema are by far the most important risk factors for the development of sAS. We here demonstrate that the genetic alterations in these two different risk groups are partially overlapping, if not identical. Our observation that a) high-level amplifications of MYC are virtually 100% specific for sAS associated with irradiation or lymphedema, and b) are present in more than 50% of cases now places this subgroup of secondary sarcomas among other sarcomas defined by recurrent high level gene amplifications, such as well-differentiated liposarcoma or neuroblastoma. Since sAS are most often observed in the setting of irradiated breast carcinomas, with a more than 1000-fold increased relative risk,10Strobbe LJ Peterse HL van Tinteren H Wijnmaalen A Rutgers EJ Angiosarcoma of the breast after conservation therapy for invasive cancer, the incidence and outcome. An unforseen sequela.Breast Cancer Res Treat. 1998; 47: 101-109Crossref PubMed Scopus (170) Google Scholar as compared with the general population, our series of sAS was heavily biased toward women and sAS of the breast. However, MYC amplifications occurred at the same relative frequency in sAS of other localizations, so we believe that MYC amplifications are a feature of sAS in general and not specific for breast sarcomas. Somewhat surprisingly, the amplification of MYC appeared not to have a major impact on tumor cell morphology or tumor cell turnover, hence the functional consequences of the MYC amplification remain yet to be established. In this multicenter retrospective study, available clinical data were insufficient to decide whether sAS with and without MYC amplification (or any other of the observed genetic changes) behaved differently in terms of prognosis. Further studies will be needed to address this relevant question. A presumably important role of MYC is not unique to sAS and has been reported also for other sarcomas, usually associated with high histological tumor grade. Interestingly, the Kaposi's sarcoma-associated herpesvirus LANA protein has been reported to stabilize and activate MYC.35Liu J Martin HJ Liao G Hayward SD The Kaposi's sarcoma-associated herpesvirus LANA protein stabilizes and activates c-Myc.J Virol. 2007; 81: 10451-10459Crossref PubMed Scopus (100) Google Scholar Increased MYC copy numbers are frequent in malignant fibrous histiocytoma of bone,36Tarkkanen M Larramendy ML Bohling T Serra M Hattinger CM Kivioja A Elomaa I Picci P Knuutila S Malignant fibrous histiocytoma of bone: analysis of genomic imbalances by comparative genomic hybridisation and C-MYC expression by immunohistochemistry.Eur J Cancer. 2006; 42: 1172-1180Abstract Full Text Full Text PDF PubMed Scopus (34) Google Scholar high grade chondrosarcoma37Morrison C Radmacher M Mohammed N Suster D Auer H Jones S Riggenbach J Kelbick N Bos G Mayerson J MYC amplification and polysomy 8 in chondrosarcoma: array comparative genomic hybridization, fluorescent in situ hybridization, and association with outcome.J Clin Oncol. 2005; 23: 9369-9376Crossref PubMed Scopus (55) Google Scholar and proximal type epitheloid sarcoma38Lualdi E Modena P Debiec-Rychter M Pedeutour F Teixeira MR Facchinetti F Dagrada GP Pilotti S Sozzi G Molecular cytogenetic characterization of proximal-type epithelioid sarcoma.Genes Chromosomes Cancer. 2004; 41: 283-290Crossref PubMed Scopus (59) Google Scholar and may be involved in the progression of myxoid liposarcomas.39Schneider-Stock R Boltze C Jager V Epplen J Landt O Peters B Rys J Roessner A Elevated telomerase activity, c-MYC-, and hTERT mRNA expression: association with tumour progression in malignant lipomatous tumours.J Pathol. 2003; 199: 517-525Crossref PubMed Scopus (36) Google Scholar MYC overexpression is also frequent in osteosarcomas40Squire JA Pei J Marrano P Beheshti B Bayani J Lim G Moldovan L Zielenska M High-resolution mapping of amplifications and deletions in pediatric osteosarcoma by use of CGH analysis of cDNA microarrays.Genes Chromosomes Cancer. 2003; 38: 215-225Crossref PubMed Scopus (137) Google Scholar where it confers resistance to methotrexate.41Scionti I Michelacci F Pasello M Hattinger CM Alberghini M Manara MC Bacci G Ferrari S Scotlandi K Picci P Serra M Clinical impact of the methotrexate resistance-associated genes C-MYC and dihydrofolate reductase (DHFR) in high-grade osteosarcoma.Ann Oncol. 2008; 19: 1500-1508Crossref PubMed Scopus (45) Google Scholar Moreover, inactivation of MYC reverses the malignant phenotype of rhabdomyosarcomas42Marampon F Ciccarelli C Zani BM Down-regulation of c-Myc following MEK/ERK inhibition halts the expression of malignant phenotype in rhabdomyosarcoma and in non muscle-derived human tumors.Mol Cancer. 2006; 5: 31Crossref PubMed Scopus (118) Google Scholar and even brief inactivation of MYC has been reported to result in sustained regression in osteosarcoma cells.43Jain M Arvanitis C Chu K Dewey W Leonhardt E Trinh M Sundberg CD Bishop JM Felsher DW Sustained loss of a neoplastic phenotype by brief inactivation of MYC.Science. 2002; 297: 102-104Crossref PubMed Scopus (541) Google Scholar These observations raise the possibility that specific manipulation of MYC may be an effective therapeutic strategy for selected sarcomas including sAS. Candidate genes for the other recurrent gene amplifications in sAS are yet to be identified. Locus 10p12.33 contains the so far uncharacterized micro-RNA mir-511-2. One of the most interesting candidate genes in locus 5q35.3 is FLT4, the gene encoding the receptor tyrosine kinase vascular endothelial growth factor receptor 3.44Galland F Karamysheva A Mattei MG Rosnet O Marchetto S Birnbaum D Chromosomal localization of FLT4, a novel receptor-type tyrosine kinase gene.Genomics. 1992; 13: 475-478Crossref PubMed Scopus (60) Google Scholar Vascular endothelial growth factor receptor 3 plays an important role both in vasculogenesis45Dumont DJ Jussila L Taipale J Lymboussaki A Mustonen T Pajusola K Breitman M Alitalo K Cardiovascular failure in mouse embryos deficient in VEGF receptor-3.Science. 1998; 282: 946-949Crossref PubMed Scopus (697) Google Scholar and angiogenesis.46Tammela T Zarkada G Wallgard E Murtomaki A Suchting S Wirzenius M Waltari M Hellstrom M Schomber T Peltonen R Freitas C Duarte A Isoniemi H Laakkonen P Christofori G Yla-Herttuala S Shibuya M Pytowski B Eichmann A Betsholtz C Alitalo K Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation.Nature. 2008; 454: 656-660Crossref PubMed Scopus (659) Google Scholar FLT4 has been frequently found mutated in hereditary lymphedema47Evans AL Brice G Sotirova V Mortimer P Beninson J Burnand K Rosbotham J Child A Sarfarazi M Mapping of primary congenital lymphedema to the 5q35.3 region.Am J Hum Genet. 1999; 64: 547-555Abstract Full Text Full Text PDF PubMed Scopus (109) Google Scholar, 48Karkkainen MJ Ferrell RE Lawrence EC Kimak MA Levinson KL McTigue MA Alitalo K Finegold DN Missense mutations interfere with VEGFR-3 signalling in primary lymphoedema.Nat Genet. 2000; 25: 153-159Crossref PubMed Scopus (536) Google Scholar and vascular endothelial growth factor receptor 3 has been found to be expressed by the vast majority of AS.49Partanen TA Alitalo K Miettinen M Lack of lymphatic vascular specificity of vascular endothelial growth factor receptor 3 in 185 vascular tumors.Cancer. 1999; 86: 2406-2412Crossref PubMed Scopus (234) Google Scholar It is of interest that the pAS did not show recurrent genetic abnormalities, at least with the techniques applied here. Available data19Gil-Benso R Lopez-Gines C Soriano P Almenar S Vazquez C Llombart-Bosch A Cytogenetic study of angiosarcoma of the breast.Genes Chromosomes Cancer. 1994; 10: 210-212Crossref PubMed Scopus (24) Google Scholar, 20Naka N Tomita Y Nakanishi H Araki N Hongyo T Ochi T Aozasa K Mutations of p53 tumor-suppressor gene in angiosarcoma.Int J Cancer. 1997; 71: 952-955Crossref PubMed Scopus (79) Google Scholar, 21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, 22Cerilli LA Huffman HT Anand A Primary renal angiosarcoma: a case report with immunohistochemical, ultrastructural, and cytogenetic features and review of the literature.Arch Pathol Lab Med. 1998; 122: 929-935PubMed Google Scholar, 23Schuborg C Mertens F Rydholm A Brosjo O Dictor M Mitelman F Mandahl N Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue.Cancer Genet Cytogenet. 1998; 100: 52-56Abstract Full Text PDF PubMed Scopus (30) Google Scholar, 24Wong KF So CC Wong N Siu LL Kwong YL Chan JK Sinonasal angiosarcoma with marrow involvement at presentation mimicking malignant lymphoma: cytogenetic analysis using multiple techniques.Cancer Genet Cytogenet. 2001; 129: 64-68Abstract Full Text Full Text PDF PubMed Scopus (27) Google Scholar, 25Zu Y Perle MA Yan Z Liu J Kumar A Waisman J Chromosomal abnormalities and p53 gene mutation in a cardiac angiosarcoma.Appl Immunohistochem Mol Morphol. 2001; 9: 24-28Crossref PubMed Scopus (32) Google Scholar, 26Mandahl N Jin YS Heim S Willen H Wennerberg J Biorklund A Mitelman F Trisomy 5 and loss of the Y chromosome as the sole cytogenetic anomalies in a cavernous hemangioma/angiosarcoma.Genes Chromosomes Cancer. 1990; 1: 315-316Crossref PubMed Scopus (28) Google Scholar, 27Kindblom LG Stenman G Angervall L Morphological and cytogenetic studies of angiosarcoma in Stewart-Treves syndrome.Virchows Arch A Pathol Anat Histopathol. 1991; 419: 439-445Crossref PubMed Scopus (39) Google Scholar, 28Van den Berg E Van Oven MW de Jong B Dam A Wiersema J Dijkhuizen T Hoekstra HJ Molenaar WM Comparison of cytogenetic abnormalities and deoxyribonucleic acid ploidy of benign, borderline malignant, and different grades of malignant soft tissue tumors.Lab Invest. 1994; 70: 307-313PubMed Google Scholar (reviewed in Ref. 21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar) suggest that pAS may also harbor recurrent aberrations, including gains on chromosome 20p (5 of 13 published cases),19Gil-Benso R Lopez-Gines C Soriano P Almenar S Vazquez C Llombart-Bosch A Cytogenetic study of angiosarcoma of the breast.Genes Chromosomes Cancer. 1994; 10: 210-212Crossref PubMed Scopus (24) Google Scholar, 21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, 23Schuborg C Mertens F Rydholm A Brosjo O Dictor M Mitelman F Mandahl N Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue.Cancer Genet Cytogenet. 1998; 100: 52-56Abstract Full Text PDF PubMed Scopus (30) Google Scholar, 25Zu Y Perle MA Yan Z Liu J Kumar A Waisman J Chromosomal abnormalities and p53 gene mutation in a cardiac angiosarcoma.Appl Immunohistochem Mol Morphol. 2001; 9: 24-28Crossref PubMed Scopus (32) Google Scholar polysomy 8 (4/13 cases),23Schuborg C Mertens F Rydholm A Brosjo O Dictor M Mitelman F Mandahl N Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue.Cancer Genet Cytogenet. 1998; 100: 52-56Abstract Full Text PDF PubMed Scopus (30) Google Scholar, 25Zu Y Perle MA Yan Z Liu J Kumar A Waisman J Chromosomal abnormalities and p53 gene mutation in a cardiac angiosarcoma.Appl Immunohistochem Mol Morphol. 2001; 9: 24-28Crossref PubMed Scopus (32) Google Scholar chromosome 7 (4/13 cases),21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, 23Schuborg C Mertens F Rydholm A Brosjo O Dictor M Mitelman F Mandahl N Cytogenetic analysis of four angiosarcomas from deep and superficial soft tissue.Cancer Genet Cytogenet. 1998; 100: 52-56Abstract Full Text PDF PubMed Scopus (30) Google Scholar, 25Zu Y Perle MA Yan Z Liu J Kumar A Waisman J Chromosomal abnormalities and p53 gene mutation in a cardiac angiosarcoma.Appl Immunohistochem Mol Morphol. 2001; 9: 24-28Crossref PubMed Scopus (32) Google Scholar and 1q (3/13 cases),21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar, 24Wong KF So CC Wong N Siu LL Kwong YL Chan JK Sinonasal angiosarcoma with marrow involvement at presentation mimicking malignant lymphoma: cytogenetic analysis using multiple techniques.Cancer Genet Cytogenet. 2001; 129: 64-68Abstract Full Text Full Text PDF PubMed Scopus (27) Google Scholar as well as losses on chromosome 5q and 16q (two cases each).21Baumhoer D Gunawan B Becker H Fuzesi L Comparative genomic hybridization in four angiosarcomas of the female breast.Gynecol Oncol. 2005; 97: 348-352Abstract Full Text Full Text PDF PubMed Scopus (26) Google Scholar Remarkably, these alterations do not overlap with the two published sAS cases and our own series. Another, more technical consideration may come from one study of a cultured angiosarcoma reported marked tumor cell heterogeneity with a diploid karyotype in about 40% of tumor cells, while the remaining cells showed a multitude of nonclonal structural and numeric alterations,27Kindblom LG Stenman G Angervall L Morphological and cytogenetic studies of angiosarcoma in Stewart-Treves syndrome.Virchows Arch A Pathol Anat Histopathol. 1991; 419: 439-445Crossref PubMed Scopus (39) Google Scholar which may be an obstacle to detect alterations in whole tissue extracts. It will be important to determine whether different technical approaches (eg, single nucleotide polymorphism arrays or analyses using microdissected tumor tissues) will reveal recurrent genetic alterations also in primary AS. In summary, we identified high level amplifications of MYC at locus 8q24.21 as a specific alteration of secondary but not primary angiosarcomas. This observation may have importance both for the molecular classification and therapy of these tumors. Download .xls (7.38 MB) Help with xls files
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