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Article

Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme

  • Authors:
    • Angelantonio Notarangelo
    • Domenico Trombetta
    • Vincenzo D'Angelo
    • Paola Parrella
    • Orazio Palumbo
    • Clelia Tiziana Storlazzi
    • Luciana Impera
    • Lucia Anna Muscarella
    • Antonella La Torre
    • Andrea Affuso
    • Vito Michele Fazio
    • Massimo Carella
    • Leopoldo Zelante
  • View Affiliations / Copyright

    Affiliations: Medical Genetics Unit, IRCCS Casa Sollievo della Sofferenza Hospital, I-71013 San Giovanni Rotondo (FG), Italy, Laboratory of Oncology, IRCCS Casa Sollievo della Sofferenza Hospital, I-71013 San Giovanni Rotondo (FG), Italy, Department of Neurosurgery, IRCCS Casa Sollievo della Sofferenza Hospital, I-71013 San Giovanni Rotondo (FG), Italy, Department of Biology, University of Bari Aldo Moro, I-70125 Bari, Italy, Biogem IRGS, I-83031 Ariano Irpino (AV), Italy
  • Pages: 717-724
    |
    Published online on: December 23, 2013
       https://doi.org/10.3892/ijo.2013.2224
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Abstract

Glioblastoma multiforme (World Health Organ­ization, grade IV astrocytoma) is the most common and most aggressive malignant primary brain tumor. We report a novel cell line, designated as ANGM-CSS, which was established from a 56-year-old male patient with a surgically removed glioblastoma multiforme. The ANGM-CSS cell line was established in vitro and characterized using histological and immunohistochemical staining, classical and molecular cytogenetic analyses, molecular studies and functional assays using a xenograft model in immunodeficient animals. ANGM-CSS was positive for CD133, nestin and vimentin proteins, whereas GFAP showed staining only in a fraction of the cells. Cytogenetic and molecular cytogenetic analysis revealed a near-tetraploid karyotype, with a modal chromosome number from 88 to 91, and additional cytogenetic abnormalities, such as the t(6;14)(p12;q11.2), t(8;10)(q24.2;q21.1) and t(5;9)(q34;p21) unbalanced translocations. Moreover, ANGM-CSS showed amplification of the MET and EGFR genes whose overexpression was observed at the mRNA level. Interestingly, ANGM-CSS is tumorigenic when implanted in immunodeficient mice, and the cells obtained from the xenografts showed the same morphology and karyotype in vitro as the original cell line. ANGM-CSS represents a biologically relevant cell line to be used to investigate the molecular pathology of glioblastoma multiforme, also to evaluate the efficacy of novel therapeutic drugs in vitro.
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1. 

Furnari FB, Fenton T, Bachoo RM, et al: Malignant astrocytic glioma: genetics, biology, and paths to treatment. Genes Dev. 21:2683–26710. 2007. View Article : Google Scholar : PubMed/NCBI

2. 

Porter K, McCarthy B, Berbaum M, et al: Conditional survival of all primary brain tumor patients by age, behavior, and histology. Neuroepidemiology. 36:230–239. 2011. View Article : Google Scholar : PubMed/NCBI

3. 

Preusser M, de Ribaupierre S, Wöhrer A, et al: Current concepts and management of glioblastoma. Ann Neurol. 70:9–21. 2011. View Article : Google Scholar

4. 

Cheng L, Wu Q, Guryanova O, et al: Elevated invasive potential of glioblastoma stem cells. Biochem Biophys Res Commun. 406:643–648. 2011. View Article : Google Scholar : PubMed/NCBI

5. 

Louis DN, Ohgaki H, Wiestler OD, et al: The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 114:97–109. 2007. View Article : Google Scholar : PubMed/NCBI

6. 

Stupp R, Mason WP, van den Bent MJ, et al: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 352:987–996. 2005. View Article : Google Scholar : PubMed/NCBI

7. 

Stupp R, Hegi ME, Mason WP, et al: Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol. 10:459–466. 2009.

8. 

Wen PY and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI

9. 

Bakir A, Gezen F, Yildiz O, et al: Establishment and characterization of a human glioblastoma multiforme cell line. Cancer Genet Cytogenet. 103:46–51. 1998. View Article : Google Scholar : PubMed/NCBI

10. 

Nuki Y, Uchinokura S, Miyata S, et al: Establishment and characterization of a new human glioblastoma cell line, NYGM. Hum Cell. 17:145–150. 2004. View Article : Google Scholar : PubMed/NCBI

11. 

Wang J, Wang X, Jiang S, et al: Establishment of a new human glioblastoma multiforme cell line (WJ1) and its partial characterization. Cell Mol Neurobiol. 27:831–843. 2007. View Article : Google Scholar : PubMed/NCBI

12. 

Fomchenko EI and Holland EC: Mouse models of brain tumors and their applications in preclinical trials. Clin Cancer Res. 12:5288–5297. 2006. View Article : Google Scholar : PubMed/NCBI

13. 

Palumbo O, Palumbo P, Palladino T, et al: A novel deletion in 2q24.1q24.2 in a girl with mental retardation and generalized hypotonia: a case report. Mol Cytogenet. 5:12012. View Article : Google Scholar : PubMed/NCBI

14. 

Macchia G, Trombetta D, Möller E, et al: FOSL1 as a candidate target gene for 11q12 rearrangements in desmoplastic fibroblastoma. Lab Invest. 92:735–743. 2012. View Article : Google Scholar : PubMed/NCBI

15. 

Scintu M, Vitale R, Prencipe M, et al: Genomic instability and increased expression of BUB1B and MAD2L1 genes in ductal breast carcinoma. Cancer Lett. 254:298–307. 2012. View Article : Google Scholar : PubMed/NCBI

16. 

Parrella P, la Torre A, Copetti M, et al: High specificity of quantitative methylation-specific PCR analysis for MGMT promoter hypermethylation detection in gliomas. J Biomed Biotechnol. 2009:5316922009. View Article : Google Scholar : PubMed/NCBI

17. 

Hibi K, Robinson CR, Booker S, et al: Molecular detection of genetic alterations in the serum of colorectal cancer patients. Cancer Res. 58:1405–1407. 1998.PubMed/NCBI

18. 

Gu TL, Deng X, Huang F, et al: Survey of tyrosine kinase signaling reveals ROS kinase fusions in human cholangiocarcinoma. PLoS One. 6:e156402011. View Article : Google Scholar : PubMed/NCBI

19. 

Singh D, Chan JM, Zoppoli P, et al: Transforming fusions of FGFR and TACC genes in human glioblastoma. Science. 337:1231–1235. 2012. View Article : Google Scholar : PubMed/NCBI

20. 

Charest A, Lane K, McMahon K, et al: Fusion of FIG to the receptor tyrosine kinase ROS in a glioblastoma with an interstitial del(6) (q21q21). Genes Chromosomes Cancer. 37:58–71. 2003. View Article : Google Scholar : PubMed/NCBI

21. 

Burfeind P, Chernicky CL, Rininsland F, et al: Antisense RNA to the type I insulin-like growth factor receptor suppresses tumor growth and prevents invasion by rat prostate cancer cells in vivo. Proc Natl Acad Sci USA. 93:7263–7268. 1996. View Article : Google Scholar : PubMed/NCBI

22. 

Shaffer LG, Slovak ML and Campbell LJ: ISCN 2009: An International System for Human Cytogenetic Nomenclature. S. Karger; Basel: 2009

23. 

Lolait SJ, Harmer JH, Auteri G, et al: Expression of glial fibrillary acidic protein, actin, fibronectin and factor VIII antigen in human astrocytomas. Pathology. 15:373–378. 1983. View Article : Google Scholar : PubMed/NCBI

24. 

Lipsky RH and Silverman SJ: Effects of mycophenolic acid on detection of glial filaments in human and rat astrocytoma cultures. Cancer Res. 47:4900–4904. 1987.PubMed/NCBI

25. 

Bocchini V, Casalone R, Collini P, et al: Changes in glial fibrillary acidic protein and karyotype during culturing of two cell lines established from human glioblastoma multiforme. Cell Tissue Res. 265:73–81. 1991. View Article : Google Scholar

26. 

Veselska R, Kuglik P, Cejpek P, et al: Nestin expression in the cell lines derived from glioblastoma multiforme. BMC Cancer. 6:322003. View Article : Google Scholar : PubMed/NCBI

27. 

Jiang Y and Uhrbom L: On the origin of glioma. Ups J Med Sci. 117:113–121. 2012. View Article : Google Scholar

28. 

Krupkova O, Loja T, Zambo I, et al: Nestin expression in human tumors and tumor cell lines. Neoplasma. 57:291–298. 2010. View Article : Google Scholar : PubMed/NCBI

29. 

Bigner DD, Bigner SH, Ponten J, et al: Heterogeneity of Genotypic and phenotypic characteristics of fifteen permanent cell lines derived from human gliomas. J Neuropathol Exp Neurol. 40:201–229. 1981. View Article : Google Scholar : PubMed/NCBI

30. 

Roversi G, Pfundt R, Moroni RF, et al: Identification of novel genomic markers related to progression to glioblastoma through genomic profiling of 25 primary glioma cell lines. Oncogene. 25:1571–1583. 2006. View Article : Google Scholar : PubMed/NCBI

31. 

Dahlback HS, Brandal P, Meling TR, et al: Genomic aberrations in 80 cases of primary glioblastoma multiforme: Pathogenetic heterogeneity and putative cytogenetic pathways. Genes Chromosomes Cancer. 48:908–924. 2009. View Article : Google Scholar

32. 

Rey JA, Bello MJ, de Campos JM, et al: Chromosomal patterns in human malignant astrocytomas. Cancer Genet Cytogenet. 29:201–221. 1987. View Article : Google Scholar : PubMed/NCBI

33. 

Bigner SH, Mark J and Bigner DD: Cytogenetics of human brain tumors. Cancer Genet Cytogenet. 47:141–154. 1990. View Article : Google Scholar : PubMed/NCBI

34. 

Kruse CA, Mitchell DH, Kleinschmidt-DeMasters BK, et al: Characterization of a continuous human glioma cell line DBTRG-05MG: growth kinetics, karyotype, receptor expression, and tumor suppressor gene analyses. In Vitro Cell Dev Biol. 28A:609–614. 1992. View Article : Google Scholar : PubMed/NCBI

35. 

Gjerset RA, Fakhrai H, Shawler DL, et al: Characterization of a new human glioblastoma cell line that expresses mutant p53 and lacks activation of the PDGF pathway. In Vitro Cell Dev Biol Anim. 31:207–214. 1995. View Article : Google Scholar : PubMed/NCBI

36. 

Cancer Genome Atlas Research Network: Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 455:1061–1068. 2008. View Article : Google Scholar : PubMed/NCBI

37. 

Uhrinova S, Uhrin D, Powers H, et al: Structure of free MDM2 N-terminal domain reveals conformational adjustments that accompany p53-binding. J Mol Biol. 350:587–598. 2005. View Article : Google Scholar : PubMed/NCBI

38. 

Esteller M, Garcia-Foncillas J, Andion E, et al: Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. N Engl J Med. 343:1350–1354. 2000. View Article : Google Scholar

39. 

Hegi ME, Diserens AC, Gorlia T, et al: MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med. 352:997–1003. 2005. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Notarangelo A, Trombetta D, D'Angelo V, Parrella P, Palumbo O, Storlazzi CT, Impera L, Muscarella LA, La Torre A, Affuso A, Affuso A, et al: Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme. Int J Oncol 44: 717-724, 2014.
APA
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C.T. ... Zelante, L. (2014). Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme. International Journal of Oncology, 44, 717-724. https://doi.org/10.3892/ijo.2013.2224
MLA
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C. T., Impera, L., Muscarella, L. A., La Torre, A., Affuso, A., Fazio, V. M., Carella, M., Zelante, L."Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme". International Journal of Oncology 44.3 (2014): 717-724.
Chicago
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C. T., Impera, L., Muscarella, L. A., La Torre, A., Affuso, A., Fazio, V. M., Carella, M., Zelante, L."Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme". International Journal of Oncology 44, no. 3 (2014): 717-724. https://doi.org/10.3892/ijo.2013.2224
Copy and paste a formatted citation
x
Spandidos Publications style
Notarangelo A, Trombetta D, D'Angelo V, Parrella P, Palumbo O, Storlazzi CT, Impera L, Muscarella LA, La Torre A, Affuso A, Affuso A, et al: Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme. Int J Oncol 44: 717-724, 2014.
APA
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C.T. ... Zelante, L. (2014). Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme. International Journal of Oncology, 44, 717-724. https://doi.org/10.3892/ijo.2013.2224
MLA
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C. T., Impera, L., Muscarella, L. A., La Torre, A., Affuso, A., Fazio, V. M., Carella, M., Zelante, L."Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme". International Journal of Oncology 44.3 (2014): 717-724.
Chicago
Notarangelo, A., Trombetta, D., D'Angelo, V., Parrella, P., Palumbo, O., Storlazzi, C. T., Impera, L., Muscarella, L. A., La Torre, A., Affuso, A., Fazio, V. M., Carella, M., Zelante, L."Establishment and genetic characterization of ANGM-CSS, a novel, immortal cell line derived from a human glioblastoma multiforme". International Journal of Oncology 44, no. 3 (2014): 717-724. https://doi.org/10.3892/ijo.2013.2224
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