The Experts below are selected from a list of 1041 Experts worldwide ranked by ideXlab platform
Thomas Ried - One of the best experts on this subject based on the ideXlab platform.
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Spectral Karyotyping analysis of human and mouse chromosomes
Nature Protocols, 2006Co-Authors: Hesed Padillanash, Linda Barenboimstapleton, Michael J Difilippantonio, Thomas RiedAbstract:Classical banding methods provide basic information about the identities and structures of chromosomes on the basis of their unique banding patterns. Spectral Karyotyping (SKY), and the related multiplex fluorescence in situ hybridization (M-FISH), are chromosome-specific multicolor FISH techniques that augment cytogenetic evaluations of malignant disease by providing additional information and improved characterization of aberrant chromosomes that contain DNA sequences not identifiable using conventional banding methods. SKY is based on cohybridization of combinatorially labeled chromosome-painting probes with unique fluorochrome signatures onto human or mouse metaphase chromosome preparations. Image acquisition and analysis use a specialized imaging system, combining Sagnac interferometer and CCD camera images to reconstruct Spectral information at each pixel. Here we present a protocol for SKY analysis using commercially available SkyPaint probes, including procedures for metaphase chromosome preparation, slide pretreatment and probe hybridization and detection. SKY analysis requires approximately 6 d.
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patterns of aneuploidy in stage iv clear cell renal cell carcinoma revealed by comparative genomic hybridization and Spectral Karyotyping
Genes Chromosomes and Cancer, 2003Co-Authors: Christian P Pavlovich, Thomas Ried, Hesed Padillanash, Danny Wangsa, Michael L Nickerson, Vera Y Matrosova, Marston W Linehan, John PhillipsAbstract:We report the use of Spectral Karyotyping (SKY) and comparative genomic hybridization (CGH) to describe the numerous genomic imbalances characteristic of stage IV clear cell renal cell carcinoma (CCRCC). SKY and CGH were performed on 10 cell lines established from nephrectomy specimens, and CGH on uncultured material from five of the primary renal tumors. The mutational status of VHL (3p25) and MET (7q31), genes implicated in renal carcinogenesis, were determined for each case. Each case showed marked aneuploidy, with an average number of copy alterations of 14.6 (+/-2.7) in the primary tumors and 19.3 (+/-4.6) in the cell lines. Both whole-chromosome and chromosome-segment imbalances were noted by CGH: consistent losses or gains included +5q23-->ter (100%), -3p14-->ter (80%), and +7 (70%). All VHL mutations and 83% of the genomic imbalances found in the primary tumors were also found in the cell lines derived from them. SKY showed many complex structural rearrangements that were undetected by conventional banding analysis in these solid tumors. All cases with VHL inactivation had 3p loss and 5q gain related primarily to unbalanced translocations between 3p and 5q. In contrast, gains of chromosome 7 resulted primarily from whole-chromosome gains and were not associated with mutations of MET. SKY and CGH demonstrated that genomic imbalances in advanced RCC were the result of either segregation errors [i.e., whole chromosomal gains and losses (7.8/case)] or chromosomal rearrangements (10.7/case), of which the majority were unbalanced translocations.
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analysis of a de novo complex chromosome rearrangement involving chromosomes 4 11 12 and 13 and eight breakpoints by conventional cytogenetic fluorescence in situ hybridization and Spectral Karyotyping
Prenatal Diagnosis, 1999Co-Authors: B Peschka, Thomas Ried, Evelin Schrock, J Leygraaf, Dagmar Hansmann, M Hansmann, Hartmut Engels, G Schwanitz, R SchubertAbstract:A complex chromosome rearrangement (CCR) with eight breakpoints resulting in four derivative chromosomes (4, 11, 12 and 13) was detected prenatally in a male fetus of a twin pregnancy. The karyotype of the female second fetus was normal. The apparently balanced de novo CCR was identified by classical cytogenetic methods and fluorescence in situ hybridization (FISH). We compared these findings with results from Spectral Karyotyping (SKY).
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prenatal diagnosis of a mosaic extra structurally abnormal chromosome by Spectral Karyotyping
Prenatal Diagnosis, 1999Co-Authors: Yi Ning, Evelin Schrock, Caroline H Laundon, Philip D Buchanan, Thomas RiedAbstract:A de novo mosaic extra structurally abnormal chromosome (ESAC) was detected in 33 per cent of cultured amniotic fluid cells from a pregnant woman. Neither Q-banding nor fluorescence in situ hybridization (FISH) employing a DNA probe for nucleolar organizer region demonstrated the presence of satellites on the ESAC. Spectral Karyotyping (SKY) was performed in this prenatal case and led to a quick and accurate determination of the ESAC as chromosome 14 in origin. The SKY finding was confirmed by conventional FISH analysis using a chromosome 14 specific painting probe. Subsequent hybridizations with a centromeric probe and a 14q subtelomeric probe were also performed to further characterize the ESAC. Absence of (TTAGGG)n sequence on the ESAC, determined postnatally, suggested it is a ring chromosome 14. Genetic counselling concerning these findings was provided to the parents who chose to continue the pregnancy. The male infant had no apparent abnormal phenotype at birth.
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molecular cytogenetic analysis of the bladder carcinoma cell line bk 10 by Spectral Karyotyping
Genes Chromosomes and Cancer, 1999Co-Authors: Hesed Padillanash, Evelin Schrock, Merryn Macville, William Nash, George M Padilla, Kathryn M Roberson, Cary N Robertson, Thomas RiedAbstract:The bladder cancer cell line BK-10 was established from a grade III–IV transitional cell carcinoma (TCC). BK-10 is near-tetraploid (±4n) and consists of two subclones with 20–25 structural aberrations. Here we report the cytogenetic analysis of BK-10 by G-banding, Spectral Karyotyping (SKY), and FISH. SKY refers to the hybridization of 24 differentially labeled chromosome painting probes and the simultaneous visualization of all human chromosomes using Spectral imaging. SKY enabled us to confirm 12 markers in BK-10 previously described by G-banding, redefine 11 aberrations, and detect 4 hidden chromosomal rearrangements, 2 of which had been identified as normal or deleted copies of chromosome 20 and 1 as a normal chromosome 3. Twenty out of 21 translocations identified were unbalanced. FISH analysis of BK-10 using chromosome arm-specific paints, centromere probes, and oncogene/tumor suppressor gene-specific probes revealed a deletion of CDKN2A (p16) in all copies of chromosome 9, a low-level amplification of MYC (five copies), and loss of one copy of TP53; detected the presence of the Y chromosome in a hidden translocation; and detected four copies of ERBB-2. A probe set for BCR and ABL verified breakpoints for all translocations involving chromosomes 9 and 22. A new karyotype presentation, “SKY-gram,” is introduced by combining data from G-banding, SKY, and FISH analysis. This study demonstrates the approach of combining molecular cytogenetic techniques to characterize fully the multiple complex chromosomal rearrangements found in the bladder cancer cell line BK-10, and to refine the chromosomal breakpoints for all translocations. Genes Chromosomes Cancer 25:53–59, 1999 Published 1999 Wiley-Liss, Inc.
Evelin Schrock - One of the best experts on this subject based on the ideXlab platform.
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Spectral Karyotyping of human mouse rat and ape chromosomes applications for genetic diagnostics and research
Cytogenetic and Genome Research, 2006Co-Authors: Evelin Schrock, P Zschieschang, Peter Obrien, Anne Helmrich, Tanja Hardt, Anja Matthaei, Karen StoutweiderAbstract:Spectral Karyotyping (SKY) is a widely used methodology to identify genetic aberrations. Multicolor fluorescence in situ hybridization using chromosome painting probes in individual colors for all metaphase chromosomes at once is combined with a unique Spectral measurement and analysis system to automatically classify normal and aberrant chromosomes. Based on countless studies and investigations in many laboratories worldwide, numerous new chromosome translocations and other aberrations have been identified in clinical and tumor cytogenetics. Thus, gene identification studies have been facilitated resulting in the dissection of tumor development and progression. For example, different translocation partners of the TEL/ETV6 transcription factor that is specially required for hematopoiesis within the bone marrow were identified. Also, the correct classification of complex karyotypes of solid tumors supports the prognostication of cancer patients. Important accomplishments for patients with genetic diseases, leukemias and lymphomas, mesenchymal tumors and solid cancers are summarized and exemplified. Furthermore, studies of disease mechanisms such as centromeric DNA breakage, DNA double strand break repair, telomere shortening and radiation-induced neoplastic transformation have been accompanied by SKY analyses. Besides the hybridization of human chromosomes, mouse Karyotyping has also contributed to the comprehensive characterization of mouse models of human disease and for gene therapy studies.
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Spectral Karyotyping and multicolor fluorescence in situ hybridization reveal new tumor specific chromosomal aberrations
Seminars in Hematology, 2000Co-Authors: Evelin Schrock, Hesed PadillanashAbstract:Abstract Spectral Karyotyping (SKY), multiple fluorescence in situ hybridization (M-FISH), cross-species color banding (Rx-FISH), multicolor chromosome banding, and other labeling techniques and strategies have been recent comprehensive technical developments in the field of molecular cytogenetics. The immediate goals of these methods are (1) to reliably characterize complex chromosomal rearrangements present in tumor karyotypes; (2) to screen for new tumor-specific chromosomal aberrations; (3) to improve genetic classification systems of different tumor types in correlation with clinical data, treatment regimens, detection of minimal residual disease, and prognosis; and (4) to identify new target regions for gene identification strategies. We present a brief overview of the different methods, including summaries of numerous published and submitted papers detailing specific cytogenetic aberrations associated with lekemias and lymphomas. To date, 640 tumor cases have been analyzed by SKY, including 410 hematologic malignancies, 146 solid tumors, and 45 mouse tumors.
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analysis of a de novo complex chromosome rearrangement involving chromosomes 4 11 12 and 13 and eight breakpoints by conventional cytogenetic fluorescence in situ hybridization and Spectral Karyotyping
Prenatal Diagnosis, 1999Co-Authors: B Peschka, Thomas Ried, Evelin Schrock, J Leygraaf, Dagmar Hansmann, M Hansmann, Hartmut Engels, G Schwanitz, R SchubertAbstract:A complex chromosome rearrangement (CCR) with eight breakpoints resulting in four derivative chromosomes (4, 11, 12 and 13) was detected prenatally in a male fetus of a twin pregnancy. The karyotype of the female second fetus was normal. The apparently balanced de novo CCR was identified by classical cytogenetic methods and fluorescence in situ hybridization (FISH). We compared these findings with results from Spectral Karyotyping (SKY).
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prenatal diagnosis of a mosaic extra structurally abnormal chromosome by Spectral Karyotyping
Prenatal Diagnosis, 1999Co-Authors: Yi Ning, Evelin Schrock, Caroline H Laundon, Philip D Buchanan, Thomas RiedAbstract:A de novo mosaic extra structurally abnormal chromosome (ESAC) was detected in 33 per cent of cultured amniotic fluid cells from a pregnant woman. Neither Q-banding nor fluorescence in situ hybridization (FISH) employing a DNA probe for nucleolar organizer region demonstrated the presence of satellites on the ESAC. Spectral Karyotyping (SKY) was performed in this prenatal case and led to a quick and accurate determination of the ESAC as chromosome 14 in origin. The SKY finding was confirmed by conventional FISH analysis using a chromosome 14 specific painting probe. Subsequent hybridizations with a centromeric probe and a 14q subtelomeric probe were also performed to further characterize the ESAC. Absence of (TTAGGG)n sequence on the ESAC, determined postnatally, suggested it is a ring chromosome 14. Genetic counselling concerning these findings was provided to the parents who chose to continue the pregnancy. The male infant had no apparent abnormal phenotype at birth.
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molecular cytogenetic analysis of the bladder carcinoma cell line bk 10 by Spectral Karyotyping
Genes Chromosomes and Cancer, 1999Co-Authors: Hesed Padillanash, Evelin Schrock, Merryn Macville, William Nash, George M Padilla, Kathryn M Roberson, Cary N Robertson, Thomas RiedAbstract:The bladder cancer cell line BK-10 was established from a grade III–IV transitional cell carcinoma (TCC). BK-10 is near-tetraploid (±4n) and consists of two subclones with 20–25 structural aberrations. Here we report the cytogenetic analysis of BK-10 by G-banding, Spectral Karyotyping (SKY), and FISH. SKY refers to the hybridization of 24 differentially labeled chromosome painting probes and the simultaneous visualization of all human chromosomes using Spectral imaging. SKY enabled us to confirm 12 markers in BK-10 previously described by G-banding, redefine 11 aberrations, and detect 4 hidden chromosomal rearrangements, 2 of which had been identified as normal or deleted copies of chromosome 20 and 1 as a normal chromosome 3. Twenty out of 21 translocations identified were unbalanced. FISH analysis of BK-10 using chromosome arm-specific paints, centromere probes, and oncogene/tumor suppressor gene-specific probes revealed a deletion of CDKN2A (p16) in all copies of chromosome 9, a low-level amplification of MYC (five copies), and loss of one copy of TP53; detected the presence of the Y chromosome in a hidden translocation; and detected four copies of ERBB-2. A probe set for BCR and ABL verified breakpoints for all translocations involving chromosomes 9 and 22. A new karyotype presentation, “SKY-gram,” is introduced by combining data from G-banding, SKY, and FISH analysis. This study demonstrates the approach of combining molecular cytogenetic techniques to characterize fully the multiple complex chromosomal rearrangements found in the bladder cancer cell line BK-10, and to refine the chromosomal breakpoints for all translocations. Genes Chromosomes Cancer 25:53–59, 1999 Published 1999 Wiley-Liss, Inc.
Hesed Padillanash - One of the best experts on this subject based on the ideXlab platform.
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Spectral Karyotyping analysis of human and mouse chromosomes
Nature Protocols, 2006Co-Authors: Hesed Padillanash, Linda Barenboimstapleton, Michael J Difilippantonio, Thomas RiedAbstract:Classical banding methods provide basic information about the identities and structures of chromosomes on the basis of their unique banding patterns. Spectral Karyotyping (SKY), and the related multiplex fluorescence in situ hybridization (M-FISH), are chromosome-specific multicolor FISH techniques that augment cytogenetic evaluations of malignant disease by providing additional information and improved characterization of aberrant chromosomes that contain DNA sequences not identifiable using conventional banding methods. SKY is based on cohybridization of combinatorially labeled chromosome-painting probes with unique fluorochrome signatures onto human or mouse metaphase chromosome preparations. Image acquisition and analysis use a specialized imaging system, combining Sagnac interferometer and CCD camera images to reconstruct Spectral information at each pixel. Here we present a protocol for SKY analysis using commercially available SkyPaint probes, including procedures for metaphase chromosome preparation, slide pretreatment and probe hybridization and detection. SKY analysis requires approximately 6 d.
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patterns of aneuploidy in stage iv clear cell renal cell carcinoma revealed by comparative genomic hybridization and Spectral Karyotyping
Genes Chromosomes and Cancer, 2003Co-Authors: Christian P Pavlovich, Thomas Ried, Hesed Padillanash, Danny Wangsa, Michael L Nickerson, Vera Y Matrosova, Marston W Linehan, John PhillipsAbstract:We report the use of Spectral Karyotyping (SKY) and comparative genomic hybridization (CGH) to describe the numerous genomic imbalances characteristic of stage IV clear cell renal cell carcinoma (CCRCC). SKY and CGH were performed on 10 cell lines established from nephrectomy specimens, and CGH on uncultured material from five of the primary renal tumors. The mutational status of VHL (3p25) and MET (7q31), genes implicated in renal carcinogenesis, were determined for each case. Each case showed marked aneuploidy, with an average number of copy alterations of 14.6 (+/-2.7) in the primary tumors and 19.3 (+/-4.6) in the cell lines. Both whole-chromosome and chromosome-segment imbalances were noted by CGH: consistent losses or gains included +5q23-->ter (100%), -3p14-->ter (80%), and +7 (70%). All VHL mutations and 83% of the genomic imbalances found in the primary tumors were also found in the cell lines derived from them. SKY showed many complex structural rearrangements that were undetected by conventional banding analysis in these solid tumors. All cases with VHL inactivation had 3p loss and 5q gain related primarily to unbalanced translocations between 3p and 5q. In contrast, gains of chromosome 7 resulted primarily from whole-chromosome gains and were not associated with mutations of MET. SKY and CGH demonstrated that genomic imbalances in advanced RCC were the result of either segregation errors [i.e., whole chromosomal gains and losses (7.8/case)] or chromosomal rearrangements (10.7/case), of which the majority were unbalanced translocations.
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Spectral Karyotyping and multicolor fluorescence in situ hybridization reveal new tumor specific chromosomal aberrations
Seminars in Hematology, 2000Co-Authors: Evelin Schrock, Hesed PadillanashAbstract:Abstract Spectral Karyotyping (SKY), multiple fluorescence in situ hybridization (M-FISH), cross-species color banding (Rx-FISH), multicolor chromosome banding, and other labeling techniques and strategies have been recent comprehensive technical developments in the field of molecular cytogenetics. The immediate goals of these methods are (1) to reliably characterize complex chromosomal rearrangements present in tumor karyotypes; (2) to screen for new tumor-specific chromosomal aberrations; (3) to improve genetic classification systems of different tumor types in correlation with clinical data, treatment regimens, detection of minimal residual disease, and prognosis; and (4) to identify new target regions for gene identification strategies. We present a brief overview of the different methods, including summaries of numerous published and submitted papers detailing specific cytogenetic aberrations associated with lekemias and lymphomas. To date, 640 tumor cases have been analyzed by SKY, including 410 hematologic malignancies, 146 solid tumors, and 45 mouse tumors.
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molecular cytogenetic analysis of the bladder carcinoma cell line bk 10 by Spectral Karyotyping
Genes Chromosomes and Cancer, 1999Co-Authors: Hesed Padillanash, Evelin Schrock, Merryn Macville, William Nash, George M Padilla, Kathryn M Roberson, Cary N Robertson, Thomas RiedAbstract:The bladder cancer cell line BK-10 was established from a grade III–IV transitional cell carcinoma (TCC). BK-10 is near-tetraploid (±4n) and consists of two subclones with 20–25 structural aberrations. Here we report the cytogenetic analysis of BK-10 by G-banding, Spectral Karyotyping (SKY), and FISH. SKY refers to the hybridization of 24 differentially labeled chromosome painting probes and the simultaneous visualization of all human chromosomes using Spectral imaging. SKY enabled us to confirm 12 markers in BK-10 previously described by G-banding, redefine 11 aberrations, and detect 4 hidden chromosomal rearrangements, 2 of which had been identified as normal or deleted copies of chromosome 20 and 1 as a normal chromosome 3. Twenty out of 21 translocations identified were unbalanced. FISH analysis of BK-10 using chromosome arm-specific paints, centromere probes, and oncogene/tumor suppressor gene-specific probes revealed a deletion of CDKN2A (p16) in all copies of chromosome 9, a low-level amplification of MYC (five copies), and loss of one copy of TP53; detected the presence of the Y chromosome in a hidden translocation; and detected four copies of ERBB-2. A probe set for BCR and ABL verified breakpoints for all translocations involving chromosomes 9 and 22. A new karyotype presentation, “SKY-gram,” is introduced by combining data from G-banding, SKY, and FISH analysis. This study demonstrates the approach of combining molecular cytogenetic techniques to characterize fully the multiple complex chromosomal rearrangements found in the bladder cancer cell line BK-10, and to refine the chromosomal breakpoints for all translocations. Genes Chromosomes Cancer 25:53–59, 1999 Published 1999 Wiley-Liss, Inc.
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analysis of b cell neoplasias by Spectral Karyotyping sky
Current Topics in Microbiology and Immunology, 1999Co-Authors: Hesed Padillanash, Evelin Schrock, Eva Hilgenfeld, Thomas RiedAbstract:B-cell neoplasias represent a heterogeneous group of diseases, including acute lymphocytic leukemia (ALL) and the broad spectrum of non-Hodgkin’s lymphomas (NHL). Conventional cytogenetic analysis has revealed specific chromosomal aberrations in ALL as well as in NHL. Spectral Karyotyping (SKY) is a novel molecular cytogenetic technique which allows the visualization of all human chromosomes in different colors, therefore greatly facilitating the recognition of chromosomal aberrations. The potential of SKY is exemplified by the fact that in our experience, 70% of the cases analyzed resulted in karyotypes where the majority of aberrations were either refined or new aberrations were detected when compared to their G-banding karyotypes. This also applies to the analysis of B-cell neoplasias. In hematologic malignancies, especially acute leukemias, specific chromosomal aberrations are of etiologic as well as diagnostic and prognostic importance. The identification of new recurrent chromosomal aberrations could therefore lead to a better characterization of disease entities or subgroups in ALL and NHL and further improve diagnosis, treatment stratification and ultimately prognosis. Interestingly, the comparison of the pattern of chromosomal aberrations in hematological neoplasias and carcinomas revealed striking differences. While about 50% of the aberrations in hematological malignancies are balanced translocations, such aberrations are exceedingly rare in epithelial cancers in which unbalanced structural and numerical aberrations prevail.
Ann Nordgren - One of the best experts on this subject based on the ideXlab platform.
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interphase fluorescence in situ hybridization and Spectral Karyotyping reveals hidden genetic aberrations in children with acute lymphoblastic leukaemia and a normal banded karyotype
British Journal of Haematology, 2001Co-Authors: Ann Nordgren, Jacqueline Schoumans, Stefan Soderhall, Magnus Nordenskjold, Elisabeth BlennowAbstract:Twenty-two cases of childhood acute lymphoblastic leukaemia (ALL) with normal G- or Q-banded karyotypes were studied by interphase fluorescence in situ hybridization (FISH) and Spectral Karyotyping. Probes detecting MLL, BCR/ABL and TEL/AML1 rearrangements were used for the interphase studies, along with centromere-specific probes from chromosomes 17 and X. In 10 patients (45%), previously undetected aberrations were demonstrable. Specific gene rearrangements and structural changes were found in six cases and numerical changes in five. Five of these aberrations have previously been reported to have an impact on prognosis. Three cases were massively hyperdiploid and, in one, the prognostically important BCR/ABL fusion was detected. In addition, a near-haploid karyotype with 27 chromosomes was found in one patient and TEL/AML1 rearrangements were detected in two cases. This study indicates that about half of childhood ALL cases with apparently normal karyotypes harbour genetic aberrations that may be detected using interphase FISH and Spectral Karyotyping.
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identification of numerical and structural chromosome aberrations in 15 high hyperdiploid childhood acute lymphoblastic leukemias using Spectral Karyotyping
European Journal of Haematology, 2001Co-Authors: Ann Nordgren, Stefan Soderhall, Magnus Nordenskjold, Filip Farnebo, Catharina Larsson, Bertil Johansson, Gosta Holmgren, Erik Forestier, Elisabeth BlennowAbstract:Spectral Karyotyping (SKY) on metaphase spreads from 15 high hyperdiploid (>51 chromosomes) childhood acute lymphoblastic leukemias (ALL), which typically display a poor chromosome morphology, was performed in order to investigate the pattern of numerical abnormalities, reveal the chromosomal origin of marker chromosomes, and identify translocations and other interchromosomal rearrangements not detected by G-banding analysis. In all cases the numerical changes could be fully characterized, and a non-random pattern of chromosomal gain was identified, with chromosomes X, 21, 14, 17, 6, 18, 4, and 10 being most frequently gained. The numerical changes had been partly misinterpreted in 12 of the 15 ALL patients using G-banding, and the present study hence emphasizes the importance of SKY in identifying such anomalies, some of which, i.e. +4 and +10, have been suggested to be prognostically important. The chromosomal origin of all marker chromosomes and of seven structural rearrangements, one of which was the prognostically important Philadelphia chromosome, could be identified. Five rearrangements [der(1)t(1;14)(q32;q21), der(2)t(2;8)(q36;?), der(3)t(2;3)(q21;?), der(8)t(8;14)(?;?), and t(9;21)(q12;q22)] have previously not been reported in ALL, emphasizing the value of SKY in identifying novel chromosomal rearrangements.
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chromosomal alterations in 15 breast cancer cell lines by comparative genomic hybridization and Spectral Karyotyping
Genes Chromosomes and Cancer, 2000Co-Authors: Soili Kytola, Ann Nordgren, Filip Farnebo, Jorma Isola, Jaana Rummukainen, Ritva Karhu, Catharina LarssonAbstract:Breast cancer cell lines have been widely used as models in functional and therapeutical studies, but their chromosomal alterations are not well known. We characterized the chromosomal aberrations in 15 commonly used human breast carcinoma cell lines (BT-474, BT-549, CAMA-1, DU4475, MCF7, MDA-MB-134, MDA-MB-157, MDA-MB-361, MDA-MB-436, MPE600, SK-BR-3, T-47D, UACC-812, UACC-893, and ZR-75-1) by comparative genomic hybridization (CGH) and Spectral Karyotyping (SKY). By CGH the most frequent gains were detected at 1q, 8q, 20q, 7, 11q13, 17q, 9q, and 16p, whereas losses were most common at 8p, 11q14–qter, 18q, and Xq. SKY revealed a multitude of structural and numerical chromosomal aberrations. Simple translocations, typically consisting of entire translocated chromosome arms, were the most common structural aberrations. Complex marker chromosomes included material from up to seven different chromosomes. Evidence for a cytogenetic aberration not previously described in breast cancer, the isoderivative chromosome, was found in two cell lines. Translocations t(8;11), t(12;16), t(1;16), and t(15;17) were frequently found, although the resulting derivative chromosomes and their breakpoints were strikingly dissimilar. The chromosomes most frequently involved in translocations were 8, 1, 17, 16, and 20. An excellent correlation was found between the number of translocation events found by SKY in the individual cell lines, and the copy number gains and losses detected by CGH, indicating that the majority of translocations are unbalanced. Genes Chromosomes Cancer 28:308–317, 2000. © 2000 Wiley-Liss, Inc.
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chromosomal alterations in 15 breast cancer cell lines by comparative genomic hybridization and Spectral Karyotyping
Genes Chromosomes and Cancer, 2000Co-Authors: Soili Kytola, Ann Nordgren, Filip Farnebo, Jorma Isola, Jaana Rummukainen, Ritva Karhu, Catharina LarssonAbstract:Breast cancer cell lines have been widely used as models in functional and therapeutical studies, but their chromosomal alterations are not well known. We characterized the chromosomal aberrations in 15 commonly used human breast carcinoma cell lines (BT-474, BT-549, CAMA-1, DU4475, MCF7, MDA-MB-134, MDA-MB-157, MDA-MB-361, MDA-MB-436, MPE600, SK-BR-3, T-47D, UACC-812, UACC-893, and ZR-75-1) by comparative genomic hybridization (CGH) and Spectral Karyotyping (SKY). By CGH the most frequent gains were detected at 1q, 8q, 20q, 7, 11q13, 17q, 9q, and 16p, whereas losses were most common at 8p, 11q14-qter, 18q, and Xq. SKY revealed a multitude of structural and numerical chromosomal aberrations. Simple translocations, typically consisting of entire translocated chromosome arms, were the most common structural aberrations. Complex marker chromosomes included material from up to seven different chromosomes. Evidence for a cytogenetic aberration not previously described in breast cancer, the isoderivative chromosome, was found in two cell lines. Translocations t(8;11), t(12;16), t(1;16), and t(15;17) were frequently found, although the resulting derivative chromosomes and their breakpoints were strikingly dissimilar. The chromosomes most frequently involved in translocations were 8, 1, 17, 16, and 20. An excellent correlation was found between the number of translocation events found by SKY in the individual cell lines, and the copy number gains and losses detected by CGH, indicating that the majority of translocations are unbalanced. Genes Chromosomes Cancer 28:308-317, 2000.
Elisabeth Blennow - One of the best experts on this subject based on the ideXlab platform.
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interphase fluorescence in situ hybridization and Spectral Karyotyping reveals hidden genetic aberrations in children with acute lymphoblastic leukaemia and a normal banded karyotype
British Journal of Haematology, 2001Co-Authors: Ann Nordgren, Jacqueline Schoumans, Stefan Soderhall, Magnus Nordenskjold, Elisabeth BlennowAbstract:Twenty-two cases of childhood acute lymphoblastic leukaemia (ALL) with normal G- or Q-banded karyotypes were studied by interphase fluorescence in situ hybridization (FISH) and Spectral Karyotyping. Probes detecting MLL, BCR/ABL and TEL/AML1 rearrangements were used for the interphase studies, along with centromere-specific probes from chromosomes 17 and X. In 10 patients (45%), previously undetected aberrations were demonstrable. Specific gene rearrangements and structural changes were found in six cases and numerical changes in five. Five of these aberrations have previously been reported to have an impact on prognosis. Three cases were massively hyperdiploid and, in one, the prognostically important BCR/ABL fusion was detected. In addition, a near-haploid karyotype with 27 chromosomes was found in one patient and TEL/AML1 rearrangements were detected in two cases. This study indicates that about half of childhood ALL cases with apparently normal karyotypes harbour genetic aberrations that may be detected using interphase FISH and Spectral Karyotyping.
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identification of numerical and structural chromosome aberrations in 15 high hyperdiploid childhood acute lymphoblastic leukemias using Spectral Karyotyping
European Journal of Haematology, 2001Co-Authors: Ann Nordgren, Stefan Soderhall, Magnus Nordenskjold, Filip Farnebo, Catharina Larsson, Bertil Johansson, Gosta Holmgren, Erik Forestier, Elisabeth BlennowAbstract:Spectral Karyotyping (SKY) on metaphase spreads from 15 high hyperdiploid (>51 chromosomes) childhood acute lymphoblastic leukemias (ALL), which typically display a poor chromosome morphology, was performed in order to investigate the pattern of numerical abnormalities, reveal the chromosomal origin of marker chromosomes, and identify translocations and other interchromosomal rearrangements not detected by G-banding analysis. In all cases the numerical changes could be fully characterized, and a non-random pattern of chromosomal gain was identified, with chromosomes X, 21, 14, 17, 6, 18, 4, and 10 being most frequently gained. The numerical changes had been partly misinterpreted in 12 of the 15 ALL patients using G-banding, and the present study hence emphasizes the importance of SKY in identifying such anomalies, some of which, i.e. +4 and +10, have been suggested to be prognostically important. The chromosomal origin of all marker chromosomes and of seven structural rearrangements, one of which was the prognostically important Philadelphia chromosome, could be identified. Five rearrangements [der(1)t(1;14)(q32;q21), der(2)t(2;8)(q36;?), der(3)t(2;3)(q21;?), der(8)t(8;14)(?;?), and t(9;21)(q12;q22)] have previously not been reported in ALL, emphasizing the value of SKY in identifying novel chromosomal rearrangements.
