Abstract
We investigated chromosomal copy number changes in ethylnitrosourea-induced and serially transplanted gliomas of the rat by flow cytometry and Comparative Genomic in situ Hybridization (CGH). CGH analysis of a primary and four transplanted tumors revealed several genomic aberrations, including whole chromosome and subchromosomal gains and losses. Gains involved rat Chromosomes (RNO) 2, 3, 4, 5, 7, 9, 11, 12, 13, and Y, whereas losses affected RNO5, 13, 20, and Y. The primary tumor exhibited gain of RNO2q31qter and gain of RNO4. While gain of RNO2 was seen in nearly all investigated passages, gain of RNO4 was apparent in the primary tumor and in passage 2 and 5 tumors. Chromosomal alterations detected as single events were restricted to the transplanted tumors and included gain of RNO3q11, 3q41qter, 5q36, 7q34qter, 9q37, 1 1q, and Y, and loss of RNO5, 13, and 20q. Flow cytometry disclosed different aneuploid cell clones in the tumors investigated. The results are discussed in analogy to findings in human glial tumors.
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References
Au W, Soukup SW, Mandybur TI (1977) Excess chromosome #4 in ethylnitrosourea-induced neurogenic tumor lines of the rat. J Natl Cancer Inst 59, 1709–1716
Barbacid M (1987) Ras genes. Annu Rev Biochem 56, 779–827
Blin N, Stafford DW (1976) A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res 3, 2303–2308
Bos JL (1989) Ras oncogenes in human cancer: a review. Cancer Res 49, 4682–4689
Britten RJ, Graham DE, Neufeld BR (1974) Analysis of repeating DNA sequences by reassociation. Methods Enzymol 29, 363–418
Collins VP (1995) Gene amplification in human gliomas. Glia 15, 289–296
De Stoppelaar JM, Suijkerbuijk RF, Van Kranen HJ, Faessen P, Mohn GR, Hoebee B (1994) Chromosomal localization of the rat N-ras protooncogene by fluorescence in situ hybridization. Cytogenet Cell Genet 67, 23–26
Du Manoir S, Speicher MR, Joos S, Schröck E, Popp S, Dönner H, Kovacs G, Robert-Nicoud M, Lichter P, Cremer T (1993) Detection of complete and partial chromosome gains and losses by comparative genomic in situ hybridization. Hum Genet 90, 590–610
Du Manoir S, Schröck E, Bentz M, Speicher MR, Joos S, Ried T, Lichter P, Cremer T (1995) Quantitative analysis of comparative genomic hybridization. Cytometry 19, 27–41
Fuller GN, Bigner SH (1992) Amplified cellular oncogenes in neoplasms of the human central nervous system. Mutat Res 276, 299–306
Haag MM, Soukup SW (1984) Association of chromosome 4 abnormalities with ethylnitrosourea-induced neuro-oncogenesis in the rat. Cancer Res 44, 784–790
Heselmeyer K, Schröck E, Du Manoir S, Biegen H, Shah K, Steinbeck R, Auer G, Ried T (1996) Gain of chromosome 3q defines the transition from severe dysplasia to invasive carcinoma of the uterine cervix. Proc Natl Acad Sci USA 93, 479–484
Islam MQ, Levan G (1987) A new fixation procedure for improved quality G-bands in routine cytogenetic work. Hereditas 107, 127–130
Kallioniemi A, Kallioniemi O-P, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D (1992) Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science 258, 818–821
Kallioniemi A, Kallioniemi O-P, Piper I, Tanner M, Stokke T, Chen L, Smith HS, Pinkel D, Gray JW, Waldman FM (1994a) Detection and mapping of amplified DNA sequences in breast cancer by comparative genomic hybridization. Proc Natl Acad Sci USA 91, 2156–2160
Kallioniemi A, Kallioniemi O-P, Piper J, Isola J, Waldman FM, Gray JW, Pinkel D (1994b) Optimizing comparative genomic hybridization for analysis of DNA sequence copy number changes in solid tumors. Genes Chromosomes Cancer 10, 231–243
Kim DH, Mohapatra G, Bollen A, Waldman FM, Feuerstein BG (1995) Chromosomal abnormalities in glioblastoma multiforme tumors and glioma cell lines detected by comparative genomic hybridization. Int J Cancer 60, 812–819
Levan G, Szpirer J, Szpirer C, Klinga K, Hanson C, Islam MQ (1991) The gene map of the Norway rat (Rattus norvegicus) and comparative mapping with mouse and man. Genomics 10, 699–718
Maggio WW (1996) Rodent glioma models. Methods Neurosci 30, 81–96
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press)
Mennel HD (1979) Transplantation of tumors of the nervous system induced by resorptive carcinogens. Neurosurg Rev 1, 37–41
Mitelman F (ed) (1995) ISCN: An International System for Human Cytogenetic Nomenclature. (Basel: S. Karger).
Romanakis K, Zankl H, Argyriou-Tirita A, Haas O (1996) DNA analysis by flow cytometry of up to nine year old methanol/acetic acid fixed samples of childhood acute lymphoblastic leukemia (ALL). In: W Hiddeman, T Büchner, (eds) Acute Leukemias V. Experimental Approaches and Management of Retractory Diseases, (Berlin, Heidelberg: Springer), pp 520–523
Schechter AL, Stern DF, Vaidyanathan L, Decker SJ, Drebin JA, Greene MI, Weinberg RA (1984) The neu oncogene: an erbB-related gene encoding a 185.000-M tumour antigen. Nature 312, 513–516
Schechter AL, Hung M-C, Vaidyanathan L, Weinberg RA, Yang-Feng TL, Francke U, Ullrich A, Coussens L (1985) The neu gene: an erbBhomologous gene distinct from and unlinked to the gene encoding the EGF receptor. Science 229, 976–978
Scherthan H, Cremer T (1994) Nonisotopic in situ hybridization in paraf-fin-embedded tissue sections. Methods Mol Genet 5, 223–236
Scherthan H, Cremer T, Arnason U, Weier H-U, Lima-de-Faria A, Frönicke L (1994) Comparative chromosome painting discloses homologous segments in distantly related mammals. Nat Genet 6, 342–347
Schlegel J, Stumm G, Mennel HD, Rüschoff J (1991) Chromosome numbers and DNA-content in intracerebrally transplanted experimental gliomas. Exp Pathol 41, 135–145
Schlegel J, Stumm G, Mennel HD (1993/94) Chemical carcinogenesis in the nervous system: past and future. Exp Toxicol Pathol 45, 455–466
Schlegel J, Scherthan H, Arens N, Stumm G, Kiessling M (1996) Detection of complex genetic alterations in human glioblastoma multiforme using comparative genomic hybridization. J Neuropathol Exp Neurol 55, 81–87
Schlegel J, Piontek G, Kühne, C, Bartels H-J, Kraus A, Kappler R, Mennel H-D (1998) Molecular genetic characterisation of intracerebrally transplanted brain tumours. Exp Toxicol Pathol, in press
Schröck E, Thiel G, Lozanova T, Du Manoir S, Meffert M-C, Jauch A, Speicher MR, Nürnberg P, Vogel S, Jänisch W, Donis-Keller H, Ried T, Witkowski R, Cremer T (1994) Comparative genomic hybridization of human malignant gliomas reveals multiple amplification sites and non-random chromosomal gains and losses. Am J Pathol 144, 1203–1217
Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602
Shi YP, Naik P, Dietrich F, Gray JW, Hanahan D, Pinkel D (1997) DNA copy number changes associated with characteristic LOH in islet cell carcinomas of transgenic mice. Genes, Chromosomes Cancer 19, 104–111
Ståhl F, Klinga-Levan K, Levan G (1997) The ratmap database, http:// ratmap.gen.gu.se
Weber RG, Sabel M, Reifenberger J, Sommer C, Oberstraß J, Reifenberger G, Kiessling M, Cremer T (1996) Characterization of genomic alterations associated with glioma progression by comparative genomic hybridization. Oncogene 13, 983–994
Yung WKA, Zhang X, Steck PA, Hung M-C (1990) Differential amplification of the tgf-α gene in human gliomas. Cancer Commun. 2, 201–205
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Kappler, R., Schlegel, J., Romanakis, K. et al. Comparative genomic in situ hybridization discloses chromosomal copy number changes in a transplanted brain tumor line of the rat (Rattus norvegicus). Mammalian Genome 9, 193–197 (1998). https://doi.org/10.1007/s003359900724
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DOI: https://doi.org/10.1007/s003359900724