Characterization of double minute chromosomes’ DNA content in a human high grade astrocytoma cell line by using comparative genomic hybridization and fluorescence in situ hybridization |
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Authors: | Michel Giollant Suzanne Bertrand Pierre Verrelle Stanislas du Manoir Thomas Ried Françoise Mornex Jean-François Doré Thomas Cremer P Malet A Tchirkov |
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Institution: | (1) Laboratoire d’Histologie, Embryologie et Cytogénétique, Faculté de Médecine, 28 Place Henri Dunant – BP 38, F-63001 Clermont-Ferrand Cedex, France Tel.: +33 73 60 80 00; Fax: +33 73 26 91 82, FR;(2) Centre Léon Bérard, INSERM, Lyon, France, FR;(3) Centre Jean Perrin et INSERM U 71, Clermont-Ferrand, France, FR;(4) National Center for Human Genome Research/NIH, Bethesda Maryland, USA, US;(5) Institute of Anthropology and Human Genetics, University of Heidelberg, Germany, DE |
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Abstract: | The presence of double minute chromosomes (dmin) in cancer cells is known to be correlated with gene amplifications. In human
high grade astrocytomas or glioblastomas, about 50% of cytogenetically characterized cases display dmin. G5 is a cell line
which has been established from a human glioblastoma containing multiple dmin. In order to identify the DNA content of these
dmin, three techniques were successively used: conventional cytogenetic analysis, comparative genomic hybridization (CGH),
and fluorescent in situ hybridization (FISH). The karyotype of G5 cells showed numerical chromosome changes (hypertriploidy),
several marker chromosomes, and multiple dmin. CGH experiments detected two strong DNA amplification areas located in 9p21-22
and 9p24, as well as an underrepresentation of chromosomes 6, 10, 11, 13, 14, and 18q. By using FISH with a chromosome 9-specific
painting probe to metaphase chromosomes of the G5 cell line, dmin were shown to contain DNA sequences originating from chromosome
9. This study demonstrates the usefulness of a combination of classical karyotyping, CGH, and FISH to identify the chromosomal
origin of amplified DNA sequences in dmin.
Received: 30 October 1994 / Revised: 25 February 1996 |
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