This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Macville, M. Articles by Ried, T. Search for Related Content PubMed PubMed Citation Articles by Macville, M. Articles by Ried, T.

Comprehensive and Definitive Molecular Cytogenetic Characterization of HeLa Cells by Spectral Karyotyping

Genome Technology Branch, National Human Genome Research Institute [M. M., E. S., H. P-N., B. M. G., T. R.], and Laboratory of Experimental Carcinogenesis, National Cancer Institute [C. K., D. Z., N. P.], NIH, Bethesda, Maryland 20892

We revisited the cytogenetic alterations of the cervical adenocarcinoma cell line HeLa through the use of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH). SKY analysis unequivocally characterized all abnormal chromosomes. Chromosomal breakpoints were primarily assigned by simultaneous assessment of SKY painted chromosomes and inverted 4,6-diamidino-2-phenylindole banding from the same cell. Twenty clonally abnormal chromosomes were found. Comparison with previously reported HeLa G-banding karyotypes revealed a remarkably stable cytogenetic constitution because 18 of 20 markers that were found were present before. The classification of 12 markers was refined in this study. Our assignment of the remaining six markers was consistent with those described in the literature.

The CGH map of chromosomal copy number gains and losses strikingly matched the SKY results and was, in a few instances, decisive for assigning breakpoints. The combined use of molecular cytogenetic methods SKY, CGH, and FISH with site-specific probes, in addition to inverted 4,6-diamidino-2-phenylindole or conventional G-banding analysis, provides the means to fully assess the genomic abnormalities in cancer cells.

Human papillomaviruses (HPVs) are frequently integrated into the cellular DNA in cervical cancers. We mapped by FISH five HPV18 integration sites: three on normal chromosomes 8 at 8q24 and two on derivative chromosomes, der(5)t(5;22;8)(q11;q11q13;q24) and der(22)t(8;22)(q24;q13), which have chromosome 8q24 material. An 8q24 copy number increase was detected by CGH. Dual-color FISH with a c-MYC probe mapping to 8q24 revealed colocalization with HPV18 at all integration sites, indicating that dispersion and amplification of the c-MYC gene sequences occurred after and was most likely triggered by the viral insertion at a single integration site. Numerical and structural chromosomal aberrations identified by SKY, genomic imbalances detected by CGH, as well as FISH localization of HPV18 integration at the c-MYC locus in HeLa cells are common and representative for advanced stage cervical cell carcinomas. The HeLa genome has been remarkably stable after years of continuous cultivation; therefore, the genetic alterations detected may have been present in the primary tumor and reflect events that are relevant to the development of cervical cancer.

This article has been cited by other articles:

				S. F. Jabbar, L. Abrams, A. Glick, and P. F. Lambert Persistence of High-Grade Cervical Dysplasia and Cervical Cancer Requires the Continuous Expression of the Human Papillomavirus Type 16 E7 Oncogene Cancer Res., May 15, 2009; 69(10): 4407 - 4414. [Abstract] [Full Text] [PDF]

				A. K. Linnemann and S. A. Krawetz Silencing by nuclear matrix attachment distinguishes cell-type specificity: association with increased proliferation capacity Nucleic Acids Res., May 1, 2009; 37(9): 2779 - 2788. [Abstract] [Full Text] [PDF]

				A. K. Linnemann, A. E. Platts, and S. A. Krawetz Differential nuclear scaffold/matrix attachment marks expressed genes Hum. Mol. Genet., February 15, 2009; 18(4): 645 - 654. [Abstract] [Full Text] [PDF]

				L. G. Acevedo, M. Bieda, R. Green, and P. J. Farnham Analysis of the Mechanisms Mediating Tumor-Specific Changes in Gene Expression in Human Liver Tumors Cancer Res., April 15, 2008; 68(8): 2641 - 2651. [Abstract] [Full Text] [PDF]

				M. Sabatino, Y. Zhao, S. Voiculescu, A. Monaco, P. Robbins, L. Karai, B. J. Nickoloff, M. Maio, S. Selleri, F. M. Marincola, et al. Conservation of Genetic Alterations in Recurrent Melanoma Supports the Melanoma Stem Cell Hypothesis Cancer Res., January 1, 2008; 68(1): 122 - 131. [Abstract] [Full Text] [PDF]

				M. L. Nguyen, R. M. Kraft, M. Aubert, E. Goodwin, D. DiMaio, and J. A. Blaho p53 and hTERT Determine Sensitivity to Viral Apoptosis J. Virol., December 1, 2007; 81(23): 12985 - 12995. [Abstract] [Full Text] [PDF]

				I. Sokolova, A. Algeciras-Schimnich, M. Song, S. Sitailo, F. Policht, B. R. Kipp, J. S. Voss, K. C. Halling, A. Ruth, W. King, et al. Chromosomal Biomarkers for Detection of Human Papillomavirus Associated Genomic Instability in Epithelial Cells of Cervical Cytology Specimens J. Mol. Diagn., November 1, 2007; 9(5): 604 - 611. [Abstract] [Full Text] [PDF]

				S. Goetze, J. Mateos-Langerak, H. J. Gierman, W. de Leeuw, O. Giromus, M. H. G. Indemans, J. Koster, V. Ondrej, R. Versteeg, and R. van Driel The Three-Dimensional Structure of Human Interphase Chromosomes Is Related to the Transcriptome Map Mol. Cell. Biol., June 15, 2007; 27(12): 4475 - 4487. [Abstract] [Full Text] [PDF]

				S. M. Gorisch, M. Wachsmuth, K. F. Toth, P. Lichter, and K. Rippe Histone acetylation increases chromatin accessibility J. Cell Sci., December 15, 2005; 118(24): 5825 - 5834. [Abstract] [Full Text] [PDF]

				S. Bergmann, B. Royer-Pokora, E. Fietze, K. Jurchott, B. Hildebrandt, D. Trost, F. Leenders, J.-C. Claude, F. Theuring, R. Bargou, et al. YB-1 Provokes Breast Cancer through the Induction of Chromosomal Instability That Emerges from Mitotic Failure and Centrosome Amplification Cancer Res., May 15, 2005; 65(10): 4078 - 4087. [Abstract] [Full Text] [PDF]

				T. Dang, S. Bao, and X.-F. Wang Human Rad9 is required for the activation of S-phase checkpoint and the maintenance of chromosomal stability Genes Cells, April 1, 2005; 10(4): 287 - 295. [Abstract] [Full Text] [PDF]

				H. S. Soifer, A. Zaragoza, M. Peyvan, M. A. Behlke, and J. J. Rossi A potential role for RNA interference in controlling the activity of the human LINE-1 retrotransposon Nucleic Acids Res., February 8, 2005; 33(3): 846 - 856. [Abstract] [Full Text] [PDF]

				A. Narezkina, K. D. Taganov, S. Litwin, R. Stoyanova, J. Hayashi, C. Seeger, A. M. Skalka, and R. A. Katz Genome-Wide Analyses of Avian Sarcoma Virus Integration Sites J. Virol., November 1, 2004; 78(21): 11656 - 11663. [Abstract] [Full Text] [PDF]

				H. Hamilton, J. Gomos, K. I. Berns, and E. Falck-Pedersen Adeno-Associated Virus Site-Specific Integration and AAVS1 Disruption J. Virol., August 1, 2004; 78(15): 7874 - 7882. [Abstract] [Full Text] [PDF]

				B. A. Van Tine, L. D. Dao, S.-Y. Wu, T. M. Sonbuchner, B. Y. Lin, N. Zou, C.-M. Chiang, T. R. Broker, and L. T. Chow Human papillomavirus (HPV) origin-binding protein associates with mitotic spindles to enable viral DNA partitioning PNAS, March 23, 2004; 101(12): 4030 - 4035. [Abstract] [Full Text] [PDF]

				A. V. Roschke, G. Tonon, K. S. Gehlhaus, N. McTyre, K. J. Bussey, S. Lababidi, D. A. Scudiero, J. N. Weinstein, and I. R. Kirsch Karyotypic Complexity of the NCI-60 Drug-Screening Panel Cancer Res., December 15, 2003; 63(24): 8634 - 8647. [Abstract] [Full Text] [PDF]

				T. Isaka, A. L. Nestor, T. Takada, and D. C. Allison Chromosomal Variations Within Aneuploid Cancer Lines J. Histochem. Cytochem., October 1, 2003; 51(10): 1343 - 1353. [Abstract] [Full Text] [PDF]

				D. Huser, S. Weger, and R. Heilbronn Packaging of Human Chromosome 19-Specific Adeno-Associated Virus (AAV) Integration Sites in AAV Virions during AAV Wild-Type and Recombinant AAV Vector Production J. Virol., April 15, 2003; 77(8): 4881 - 4887. [Abstract] [Full Text] [PDF]

				D. Huser and R. Heilbronn Adeno-associated virus integrates site-specifically into human chromosome 19 in either orientation and with equal kinetics and frequency J. Gen. Virol., January 1, 2003; 84(1): 133 - 137. [Abstract] [Full Text] [PDF]

				M. A. Harding, K. C. Arden, J. W. Gildea, J. J. Gildea, E. J. Perlman, C. Viars, and D. Theodorescu Functional Genomic Comparison of Lineage-related Human Bladder Cancer Cell Lines with Differing Tumorigenic and Metastatic Potentials by Spectral Karyotyping, Comparative Genomic Hybridization, and a Novel Method of Positional Expression Profiling Cancer Res., December 1, 2002; 62(23): 6981 - 6989. [Abstract] [Full Text] [PDF]

				D. Huser, S. Weger, and R. Heilbronn Kinetics and Frequency of Adeno-Associated Virus Site-Specific Integration into Human Chromosome 19 Monitored by Quantitative Real-Time PCR J. Virol., June 27, 2002; 76(15): 7554 - 7559. [Abstract] [Full Text] [PDF]

				P. Platzer, M. B. Upender, K. Wilson, J. Willis, J. Lutterbaugh, A. Nosrati, J. K. V. Willson, D. Mack, T. Ried, and S. Markowitz Silence of Chromosomal Amplifications in Colon Cancer Cancer Res., February 1, 2002; 62(4): 1134 - 1138. [Abstract] [Full Text] [PDF]

				X.-b. Zhong, P. M. Lizardi, X.-h. Huang, P. L. Bray-Ward, and D. C. Ward Visualization of oligonucleotide probes and point mutations in interphase nuclei and DNA fibers using rolling circle DNA amplification PNAS, March 27, 2001; 98(7): 3940 - 3945. [Abstract] [Full Text] [PDF]

				X. Sastre-Garau, M. Favre, J. Couturier, and G. Orth Distinct patterns of alteration of myc genes associated with integration of human papillomavirus type 16 or type 45 DNA in two genital tumours J. Gen. Virol., August 1, 2000; 81(8): 1983 - 1993. [Abstract] [Full Text]

				H. Tsurui, H. Nishimura, S. Hattori, S. Hirose, K. Okumura, and T. Shirai Seven-color Fluorescence Imaging of Tissue Samples Based on Fourier Spectroscopy and Singular Value Decomposition J. Histochem. Cytochem., May 1, 2000; 48(5): 653 - 662. [Abstract] [Full Text]

				H. M.J. Kerstens, J. C.M. Robben, P. J. Poddighe, W. J.G. Melchers, H. Boonstra, P. C.M. de Wilde, M. V.E. Macville, and A. G.J.M. Hanselaar AgarCyto: A Novel Cell-processing Method for Multiple Molecular Diagnostic Analyses of the Uterine Cervix J. Histochem. Cytochem., May 1, 2000; 48(5): 709 - 718. [Abstract] [Full Text]

				L. A. Koopman, K. Szuhai, J. D. H. van Eendenburg, V. Bezrookove, G. G. Kenter, E. Schuuring, H. Tanke, and G. Jan Fleuren Recurrent Integration of Human Papillomaviruses 16, 45, and 67 Near Translocation Breakpoints in New Cervical Cancer Cell Lines Cancer Res., November 1, 1999; 59(21): 5615 - 5624. [Abstract] [Full Text] [PDF]