Abstract
Alterations of genomic DNA in eight primary astrocytic tumors and two glioma cell lines were examined using a recently developed two-dimensional gel electrophoresis method called restriction landmark genomic scanning (RLGS). RLGS allows us to detect amplifications, deletions, and methylation in genomic DNA in one procedure without requiring any polymorphic markers. Approximately 2000 spots (landmark sites) in tumor specimens were compared with those in normal brain tissue. The 10 spots with intensified signal were reproducibly detected in at least 50% of primary tumors, implying amplification of corresponding DNA sequences. Conversely, 12 spots with reduced signal were observed in more than 50% of all tumors, suggesting inactivation by allelic loss, homozygous deletion, or CpG island methylation. These results suggest that common genetic alterations are closely correlated with the genesis or progression of human gliomas.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahmed Rasheed BK, McLendon RE, Friedman HS et al. (1995) Chromosome 10 deletion mapping in human gliomas: a common deletion region in 10q25. Oncogene 10:2243–2246
El-Azouzi M, Chung RY, Farmer GE et al. (1989) Loss of distinct regions on the short arm of chromosome 17 associated with tumorigenesis of human astrocytomas. Proc Natl Acad Sci USA 86:7186–7190
Fults D, Brockmeyer D, Tullous MW et al. (1992) p53 mutation and loss of heterozygosity on chromosomes 17 and 10 during human astrocytoma progression. Cancer Res 52:674–679
Henson JW, Schnitker BL, Correa KM et al. (1994) The retinoblastoma gene is involved in malignant progression of astrocytomas. Ann Neurol 36:714–721
Hirvonen HE, Salonen R, Sandberg MM et al. (1994) Differential expressio of myc, max and RB1 genes in human gliomas and glioma cell lines. Br J Cancer 69:16–25
Kinzler KW, Bigner SH, Bigner DD et al. (1987) Identification of an amplified, highly expressed gene in a human glioma. Science 236:70–73
Kumabe T, Sohma Y, Kayama T et al. (1992) Amplification of alpha-platelet-derived growth factor receptor gene lacking an exon coding for a portion of the extracellular region in a primary brain tumor of glial origin. Oncogene 7:627–633
Liberman TA, Nusbaum HR, Razon N et al. (1985) Amplification, enhanced expression and possible rearrangement of EGF receptor gene in primary human brain tumours of glial origin. Nature 313:144–147
Nakamura M, Konishi N, Hiasa Y et al. (1996) Immunohistochemical detection of CDKN2, retinoblastoma and p53 gene products in primary astrocytic tumors. Int J Oncol 8:889–893
Reifenberger G, Liu L, Ichimura K et al. (1993) Amplification and overexpression of the MDM2 gene in a subset of human malignant gliomas without p53 mutations. Cancer Res 53:2736–2739
Reifenberger G, Reifenberger G, Ichimura K et al. (1995) Amplification at 12q13–14 in human malignant gliomas is frequently accompanied by loss of heterozygosity at loci proximal and distal to the amplification site. Cancer Res 55:731–734
Schmidt EE, Ichimura K, Reifenberger G et al. (1994) CDKN2 (p16/MTS1) gene deletion or CDK4 amplification occurs in the majority of glioblastomas. Cancer Res 54:6321–6324
Sonoda Y, Iizuka M, Yasuda J et al. (1995) Loss of heterozygosity at 11p15 in malignant glioma. Cancer Res 55:2166–2168
Tsuzuki T, Tsunoda S, Sakaki T et al. (1996) Alterations of RB, p53, p16 (CDKN2), and p15 genes in human astrocytomas. Cancer 78:287–293
Ueki K, Ono Y, Henson JW et al. (1996) CDKN2/p16 or RB alterations occur in the majority of glioblastomas and are inversely correlated. Cancer Res 56:150–153
Venter DJ, Thomas DGT (1991) Multiple sequential molecular abnormalities in the evolution of human gliomas. Br J Cancer 63:753–757
Wong AJ, Bigner SH, Bigner DD et al. (1987) Increased expression of the epidermal growth factor receptor gene in malignant gliomas is invariably associated with gene amplification. Proc Natl Acad Sci USA 84:6899–6903
Hatada I, Hayashizaki Y, Hirotsune S et al. (1991) A genomic scanning method for higher organisms using restriction sites as landmarks. Proc Natl Acad Sci USA 88:9523–9527
Nagai H, Hirotsune S, Komatsubara H et al. (1993) Genomic analysis of human hepatocellular carcinomas using restriction landmark genomic scanning. Cancer Detect Prev 17:399–404
Asakawa J, Kuick R, Neel JV et al. (1994) Genetic variation detected by quantitative analysis of end-labeled genomic DNA fragments. Proc Natl Acad Sci USA 91:9052–9056
Hayashizaki Y, Hirotsune S, Okazaki Y et al. (1994) A genetic linkage map of the mouse using restriction landmark genomic scanning (RLGS). Genetics 138:1207–1238
Hayashizaki Y, Shibata H, Hirotsune S et al. (1994) Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method. Nat Genet 6:33–40
Hirotsune S, Hatada I, Komatsubara H et al. (1992) New approach for detection of amplification in cancer DNA using restriction landmark genomic scanning. Cancer Res 52:3642–3647
Hirotsune S, Hirose K, Kataoka H et al. (1994) Spot mapping on the standard profile of restriction landmark genomic scanning (RLGS) of sorted chromosome 20 using methlation-insensitive enzyme. Genomics 24:593–596
Kawai J, Hirose K, Fushiki S et al. (1994) Comparison of DNA methylation patterns among mouse cell lines by restriction landmark genomic scanning. Mol Cell Biol 14:7421–7427
Kuick R, Asakawa J, Neel JV et al. (1995) High yield of restriction fragment length polymorphisms in two-dimensional separations of human genomic DNA. Genomics 25:345–353
Nagai H, Ponglikitmongkol M Mita E et al. (1994) Aberration of genomic DNA in association with human hepatocellular carcinomas detected by 2-dimensional gel analysis. Cancer Res 54:1545–1550
Nagai H, Pongliktmongkol M, Kim YS et al. (1995) Cloning of Not I-cleaved genomic DNA fragments appearing as spots in 2D gel electrophoresis. Biochem Biophys Res Commun 213:258–265
Nagai H, Tsumura H, Ponglikimongkol M, et al. (1995) Genomic aberrations in human hepatoblastomas detected by 2-dimensional gel analysis. Cancer Res 55:4549–4551
Okazaki Y, Hirose K, Hirotsune S et al. (1995) Direct detection and isolation of restriction landmark genomic scanning (RLGS) spot DNA markers tightly linked to a specific trait by using the RLGS spot-bombing method. Proc Natl Acad Sci USA 92:5610–5614
Kleihues P, Burger PC, Scheithauer BW (1993) Histological typing of tumours of the central nervous system. Springer-Verlag, Berlin
Blin N, Stafford DW (1976) A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res 3:2303–2308
Jones PA (1996) DNA methylation errors and cancer. Cancer Res 56:2463–2467
Gonzalez-Zulueta M, Bender CM, Yang AS et al. (1995) Methylation of the 5′ CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing. Cancer Res 55:4531–4535
Makos M, Nelkin BD, Chazin VR et al. (1993) DNA hypermethylation is associated with 17p allelic loss in neural tumors. Cancer Res 53:2715–2718
Merlo A, Herman JG, Mao L et al. (1995) 5′ CpG island methylation is associated with transcriptional silencing of the tumor suppressor p16/CDKN2/MTS1 in human cancers. Nature Med 1: 686–692
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nakamura, M., Konishi, N., Tsunoda, S. et al. Genomic alterations of human gliomas detected by restriction landmark genomic scanning. Brain Tumor Pathol 14, 13–17 (1997). https://doi.org/10.1007/BF02478863
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02478863