Metaphase Chromosome Anomaly: Association with Drug Resistance and Cell-Specific Products

Biedler, June L.; Spengler, Barbara A.

doi:10.1126/science.942798

Cited by 386 publications

(126 citation statements)

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“…A number of human tumors (1)(2)(3), in particular neuroblastomas (4)(5)(6), exhibit homogeneously staining chromosomal regions (HSRs) and extrachromosomal double minute objects (DMs) which are probably cytological manifestations of amplified genes (see ref. 7 for a review).…”

mentioning

confidence: 99%

Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes.

Kanda¹,

Schreck²,

Alt³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

103

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authors request that the following correction be noted. All hybrids described in Fig. 5 contained at least part of the human X chromosome, and the box for each hybrid in the column corresponding to the human X should have been darkened. This clerical error does not alter any conclusions in the paper.Proc. Natt Acad. Sci. USA Vol. 80, pp. A number of human tumors (1-3), in particular neuroblastomas (4-6), exhibit homogeneously staining chromosomal regions (HSRs) and extrachromosomal double minute objects (DMs) which are probably cytological manifestations of amplified genes (see ref. 7 for a review). These H-SRs and DMs are often observed in tumor tissue removed prior to institution of therapy, and it seems reasonable to suggest that they may contain genetic information responsible for some of the phenotypic properties of the tumor cells. This hypothesis can be tested by isolation and characterization of DNA within the amplified chromosomal regions. In one study (8,9), DMs of a rat adrenal tumor cell line were isolated by differential centrifugation and cloned in a A bacteriophage. We have utilized a different approach to isolate sequences specific for the HSRs of a human neuroblastoma cell, exploiting the increased size imparted to metaphase chromosomes by these HSRs.The human neuroblastoma cell line IMR-32, which was established from tissue of a patient who had not been exposed to any chemotherapy (10), constitutes an excellent system for analysis of HSR-specific sequences. IMR-32 cells, which exhibit some karyotypic heterogeneity, possess, in addition to a normal chromosome 1, two abnormal chromosomes 1 in which an HSR is inserted in the short arm (in the middle of band lp3). The HSR-containing chromosome 1 is 50% larger than the normal chromosome 1 (11) and hence is easily separable from it and all other, smaller, human chromosomes by fluorescence-activated sorting of isolated IMR-32 metaphase chromosomes after they have been stained with a DNA-specific fluorochrome such as 33258 Hoechst (12).From the DNA of a few million metaphase chromosomes enriched for the HSR-containing chromosome I by flow sorting, we have prepared recombinant libraries in the Charon phage 21A (13) from which cloned sequences specific for the HSR have been identified both by DNA blotting (14) and by in situ hybridization (15, 16). These cloned sequences provide a unique opportunity to study the composition, formation, and functional significance of the amplified sequences in this and other neuroblastoma cell lines.MATERIAL AND METHODS IMR-32 neuroblastoma cells, originally obtained from the American Type Tissue Culture Collection and maintained in culture for several months, were used in these experiments. Cells were cultured in Ham F10 medium supplemented with 10% fetal calf serum. Cytological metaphase chromosome preparations were stained with quinacrine or chromomycin A3 plus methyl green as described (17).Nonconfluent cells were trapped at metaphase by treatment with Colcemid (45 ng/ml) for 20 hr. Cells were detached by shaking, d...

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mentioning

confidence: 99%

Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes.

Kanda¹,

Schreck²,

Alt³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

103

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“…The cytogenetic analysis of the 17 cell cultures derived from the tumors showed that about one-third of the tumor lines exhibited sizeable chromosome regions with a stereotype, light staining, usually referred to as HSR (homogeneously staining regions) and indicative of increased copy numbers of chromatin segments (`gene ampli®cation'; Biedler and Spengler, 1976;Levan et al, 1976;Nunberg et al, 1978). Information about ampli®cation, the location of the HSR regions, and other characteristic cytogenetic aberrations is given in Table 1.…”

Section: Cytogenetic Analysis and Cgh-analysis Of The Lb32t Tumormentioning

confidence: 99%

Amplification and overexpression of the hepatocyte growth factor receptor (HGFR/MET) in rat DMBA sarcomas

et al. 1999

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In the present study subcutaneous ®brosarcomas were induced by the carcinogen 7,12-dimethylbenz(a)anthracene (DMBA) in rats from F1 generation cross breedings of two dierent inbred strains. Comparative genomic hybridization (CGH) analysis, which allows detection of DNA sequence copy changes, was applied to one of the tumors and it was found that there were increased copy numbers of sequences at chromosome 4q12-q21 in this tumor. We have previously determined that the loci for the hepatocyte growth factor (Hgf) and hepatocyte growth factor receptor (Hgfr/Met), a protooncogene, are situated in this particular chromosome region. Using probes for the two genes in FISH (¯uorescence in situ hybridization) and in Southern blots we found that the Hgfr/Met gene was ampli®ed in ®ve of the 19 sarcomas studied, and that the Hgf gene was coampli®ed in two of them. Northern and Western blots and tyrosine phosphorylation analysis showed that the HGF receptor was overexpressed and functional in all ®ve tumors, as well as in two additional tumors. In summary, both ampli®cation and overexpression of the Hgfr/Met gene was found in about 25% of DMBAinduced experimental rat sarcomas, and HGF receptor overexpression alone was seen in two additional tumors. Possibly this re¯ects an involvement in paracrine or autocrine stimulation of growth and invasiveness by HGF. Our ®nding could provide a rodent model system to increased knowledge about causality and therapy, which may be applicable to the sizeable fraction of human musculoskeletal tumors displaying MET overexpression.

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“…Lan-1 and Lan-6 were obtained from Dr Seeger (Childrens Hospital Los Angeles, Los Angeles, USA). SK-N-BE(2) were obtained from Dr Biedler (Biedler and Spengler, 1976) and IMR32 were purchased from American Type Culture Collection (Rockville, MD, USA). Cells were grown in Dulbecco's modified eagle's medium (DMEM) (Irvine Scientific, Irvine, CA, USA) supplemented with 10% fetal bovine serum, glutamine and penicillin/streptomycin (GIBCO/BRL, Grand Island, NY, USA).…”

Section: Cell Culturementioning

confidence: 99%

The RET and TRKA pathways collaborate to regulate neuroblastoma differentiation

Peterson

Bogenmann

2004

Oncogene

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Neuroblastoma (NB) is a childhood cancer that arises in the adrenal gland and often shows differentiated neuronal and glial elements. The RET receptor signal pathway is functional in most NB, while loss of nerve growth factor (NGF) receptor (trkA) gene expression correlates with an aggressive phenotype. Thus, we hypothesized that the RET and TRKA signal pathways collaborate to instruct NB differentiation, reminiscent of normal neuronal maturation. Here, we demonstrate that activation of the RET receptor by glial cell line-derived neurotrophic factor (GDNF) increases expression of the RET receptor complex in a panel of malignant human NB cell lines, indicative of a positive feedback mechanism. GDNF also induces growth cessation concomitant with an arrest of cells in the G 0 /G 1 phase of the cell cycle. Furthermore, GDNF synergizes with ciliary neurotrophic factor (CNTF) to enhance TRKA receptor expression, thereby strengthening the NGF-mediated differentiation signal. Differentiated NB cells downregulate expression of the amplified N-myc gene, concurrent with the arrest of cell proliferation, while expressing neuron-specific markers (i.e., SCG10). Interestingly, maintenance of differentiated NB cells in culture is independent of the trophic activity of GDNF, but depends on TRKA signaling, thereby reenacting the differentiation of normal sympathoadrenal (SA) progenitor cells.

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Metaphase Chromosome Anomaly: Association with Drug Resistance and Cell-Specific Products

Cited by 386 publications

References 8 publications

Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes.

Isolation of amplified DNA sequences from IMR-32 human neuroblastoma cells: facilitation by fluorescence-activated flow sorting of metaphase chromosomes.

Amplification and overexpression of the hepatocyte growth factor receptor (HGFR/MET) in rat DMBA sarcomas

The RET and TRKA pathways collaborate to regulate neuroblastoma differentiation

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