Multiple sites of highly amplified DNA sequences detected by molecular cytogenetic analysis in HS-RMS-2, a new pleomorphic rhabdomyosarcoma cell line

Division of Human Health and Medical Science, Graduate School of Kuroshio Science, Kochi University, Nankoku, Kochi 783-8505, Japan.
American Journal of Cancer Research (Impact Factor: 4.17). 01/2011; 2(2):141-52.
Source: PubMed


A molecular cytogenetic analysis was performed on HS-RMS-2, a cell line established in this laboratory from a rare pleomorphic type of rhabdomyosarcoma. G-banding and multicolor-FISH analyses revealed that the cells have a complex chromosomal composition. Comparative genomic in situ hybridization (CGH) detected eight highly amplified regions at 1p36.1-p36.2, 1p31-p32, 1q21-q31, 8q12-q21, 8q24-qter, 11q12-q13, 12q13-q14 and 18q12-q22, suggesting the co-existence of multiple amplified oncogenes in these tumor cells. Reverse chromosome painting, using a probe regenerated by microdissection of a long marker chromosome, revealed the native location of three of eight possible genes to be on chromosomes 1p31-32, 12q14 and 18q21. FISH using BAC and cosmid probes revealed amplification of JUN (1p31), MYC (8q24), CCND1 (11q13), INT2 (11q13.3), MDM2 (12q14.3-q15) and MALT (18q21). These findings indicate that at least eight amplified oncogenes may contribute to the pathogenesis of a rare pleomorphic type of rhabdomyosarcoma. This new cell line should prove useful for in vitro preclinical studies of molecularly targeted therapies.

Download full-text


Available from: Takahiro Taguchi
  • Source
    • "UK). Random prime labeling of the probe DNA from PCR products was performed with fluorescein-12-dUTP (F-dUTP) or cyanine-3-duTP (Cy3-dUTP) in accordance with the kit protocol (Fig. 2) (Invitrogen, Tokyo, |apan). FISH analysis was carried out as previously reported (Takaoka et a1.,2012), with slight modifications. Briefly, the metaphase preparations were denatured in 70o/o formamide/2xSSC (saline-sodium citrate) at73"C for 3 min. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Molecular cytogenetic investigation based on fluorescence in situ hybridization (FISH) of the scleractinian coral, Echinophyllia aspera Ellis and Solander 1788, which is commonly found along temperate coasts in Japan (30-35°N) and in coastal waters in the Indian and Pacific oceans, was performed. FISH was applied in the study of this coral, using E. aspera embryo (about 9-12 hours after artificial fertilization; prawn chip stage). By genomic DNA hybridization (GDH) using DNA that was extracted from E. aspera embryos, we have succeeded in displaying the characteristic and distinct heterochromatin distribution, especially on telomeric regions of chromosomes, which may facilitate the classification of corals. FISH mapping of rRNA genes (rDNAs) was successfully carried out with the probe generated by PCR amplification using rRNA gene primers and revealed that extraordinary amplification of rDNA occurred in one of the homologous chromosomes similar to that in a homogeneous staining region (hsr) that is sometimes seen in human cancer cells. The presence of telomere sequences, (TTAGGG)n, in all chromosomes was visualized and demonstrated that this coral had the same sequences as humans. Based on these results obtained by FISH, we proposed the karyotype of this coral (2n = 28). Furthermore, we found that the telomeric heterochromatin in this coral contained the human satellite III DNA motif sequence (TTCCA)n, which is located on human chromosome 9 centromere. Taken together, these data suggest that karyotyping, rRNA gene mapping and heterochromatin motif sequences are useful tools for exploring the process of chromosome evolution, and phylogenetics of scleractinian coral.
    Full-text · Article · Jan 2014
  • [Show abstract] [Hide abstract]
    ABSTRACT: Rare diseases typically affect fewer than 200 000 patients annually, yet because thousands of rare diseases exist, the cumulative impact is millions of patients worldwide. Every form of childhood cancer qualifies as a rare disease-including the childhood muscle cancer, rhabdomyosarcoma (RMS). The next few years promise to be an exceptionally good era of opportunity for public-private collaboration for rare and childhood cancers. Not only do certain governmental regulation advantages exist, but these advantages are being made permanent with special incentives for pediatric orphan drug-product development. Coupled with a growing understanding of sarcoma tumor biology, synergy with pharmaceutical muscle disease drug-development programs, and emerging publically available preclinical and clinical tools, the outlook for academic-community-industry partnerships in RMS drug development looks promising.Oncogene advance online publication, 13 May 2013; doi:10.1038/onc.2013.129.
    No preview · Article · May 2013 · Oncogene
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence. Despite intergroup clinical trials conducted in Europe and North America, outcomes for high risk patients with this disease have not significantly improved in the last several decades, and survival of metastatic or relapsed disease remains extremely poor. Accrual into new clinical trials is slow and difficult, so in vitro cell-line research and in vivo xenograft models present an attractive alternative for preclinical research for this cancer type. Currently, 30 commonly used human RMS cell lines exist, with differing origins, karyotypes, histologies, and methods of validation. Selecting an appropriate cell line for RMS research has important implications for outcomes. There are also potential pitfalls in using certain cell lines including contamination with murine stromal cells, cross-contamination between cell lines, discordance between the cell line and its associated original tumor, imposter cell lines, and nomenclature errors that result in the circulation of two or more presumed unique cell lines that are actually from the same origin. These pitfalls can be avoided by testing for species-specific isoenzymes, microarray analysis, assays for subtype-specific fusion products, and short tandem repeat analysis.
    Full-text · Article · Jul 2013 · Frontiers in Oncology
Show more

We use cookies to give you the best possible experience on ResearchGate. Read our cookies policy to learn more.