Telomerase activity in primary and secondary glioblastomas multiforme as a novel molecular tumor marker

Kunyu Harada M.D., Ph.D. 1 , Kaoru Kurisu M.D., Ph.D. 1 , Hidetoshi Tahara Ph.D. 1 , Eiji Tahara Ph.D. 1 , Toshinori Ide Ph.D. 1 , and Eiichi Tahara M.D., Ph.D. 1
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  • 1 Department of Neurosurgery, Kanbara Hospital; and Departments of Neurosurgery and Cellular and Molecular Biology, and First Department of Pathology, School of Medicine, Hiroshima University, Hiroshima, Japan
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Object. Telomerase activity is responsible for cell immortality. To examine the role of telomerase in the carcinogenesis of human glioblastomas multiforme (GBMs), the authors studied telomerase activity, telomerase component expression, and telomere lengths in 42 GBM samples.

Methods. In all samples, EGFR and MDM2 amplifications and overexpressions were examined using Southern and Northern blot analyses. The p53 mutation was analyzed using polymerase chain reaction—single strand conformational polymorphism and by direct sequence analysis. Specimens of tissues were immunostained with p53, EGFR, and MDM2 antibodies. Allelic loss on chromosomes 17p and 10 was assessed by loss of heterozygosity (LOH) assays. Telomerase activity, expression of its components (human telomerase reverse transcriptase [hTERT], human telomerase RNA component [hTERC], and telomerase-associated protein [TEP1]), and telomere lengths were analyzed using the telomeric repeat amplification protocol (TRAP)—hybridization protection assay, reverse transcription—polymerase chain reaction, and Southern blot analysis. According to the results of assessments of EGFR and MDM2 amplifications, p53 mutation, LOHs in chromosomes 17p and 10, and the clinical course of the disease, the 42 samples were classified into 22 primary and 20 secondary glioblastomas.

Twenty-six (61.9%) of all 42 samples demonstrated detectable telomerase activity during the TRAP assay. Secondary GBMs displayed significantly higher levels of telomerase activity and hTERT expression than primary GBMs. Tumors with a p53 gene mutation demonstrated significantly higher telomerase activity than those without a p53 mutation. Four samples with a codon 175 mutation demonstrated an exceptionally high amount of telomerase activity. In secondary GBMs, the increase in telomerase activity and the hTERT expression level correlated with the increased frequency of p53 mutations. There was no significant difference in telomere length between primary and secondary GBMs.

Conclusions. These results suggest that telomerase activity and p53 mutations both play important roles in the multistep carcinogenesis of GBMs. Telomerase activity and hTERT expression may be considered as novel distinctive factors in human GBMs.

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Contributor Notes

Address reprint requests to: Kunyu Harada, M.D., Ph.D., Department of Neurosurgery, Kanbara Hospital, Akasaka-cho, Akasaka 1313, Fukuyama-City, Hiroshima, 720–0843 Japan. email: neuro007@urban.ne.jp.
  • 1.

    Arnold LJ Jr, , Hammond PW, & Wiese WA, et al: Assay formats involving acridinium-ester-labeled DNA probes. Clin Chem 35:15891594, 1989 Arnold LJ Jr, Hammond PW, Wiese WA, et al: Assay formats involving acridinium-ester-labeled DNA probes. Clin Chem 35:1589–1594, 1989

    • Search Google Scholar
    • Export Citation
  • 2.

    Brown T, , Aldous W, & Lance R, et al: The association between telomerase, p53, and clinical staging in colorectal cancer. Am J Surg 175:364366, 1998 Brown T, Aldous W, Lance R, et al: The association between telomerase, p53, and clinical staging in colorectal cancer. Am J Surg 175:364–366, 1998

    • Search Google Scholar
    • Export Citation
  • 3.

    Chin L, , Artandi SE, & Shen Q, et al: p53 deficiency rescues the adverse effects of telomere loss and cooperates with telomere dysfunction to accelerate carcinogenesis. Cell 97:527538, 1999 Chin L, Artandi SE, Shen Q, et al: p53 deficiency rescues the adverse effects of telomere loss and cooperates with telomere dysfunction to accelerate carcinogenesis. Cell 97:527–538, 1999

    • Search Google Scholar
    • Export Citation
  • 4.

    Counter CM, , Avilion AA, & LeFeuvre CE, et al: Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:19211929, 1992 Counter CM, Avilion AA, LeFeuvre CE, et al: Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:1921–1929, 1992

    • Search Google Scholar
    • Export Citation
  • 5.

    Counter CM, , Gupta J, & Harley CB, et al: Telomerase activity in normal leukocytes and in hematologic malignancies. Blood 85:23152320, 1995 Counter CM, Gupta J, Harley CB, et al: Telomerase activity in normal leukocytes and in hematologic malignancies. Blood 85:2315–2320, 1995

    • Search Google Scholar
    • Export Citation
  • 6.

    Feng J, , Funk WD, & Wang SS, et al: The RNA component of human telomerase. Science 269:12361241, 1995 Feng J, Funk WD, Wang SS, et al: The RNA component of human telomerase. Science 269:1236–1241, 1995

    • Search Google Scholar
    • Export Citation
  • 7.

    Gaidano G, , Volpe G, & Pastore C, et al: Detection of BCL-6 rearrangements and p53 mutations in malt-lymphomas. Am J Hematol 56:206213, 1997 Gaidano G, Volpe G, Pastore C, et al: Detection of BCL-6 rearrangements and p53 mutations in malt-lymphomas. Am J Hematol 56:206–213, 1997

    • Search Google Scholar
    • Export Citation
  • 8.

    Goh HS, , Yao J, & Smith DR: p53 point mutation and survival in colorectal cancer patients. Cancer Res 55:52175221, 1995 Goh HS, Yao J, Smith DR: p53 point mutation and survival in colorectal cancer patients. Cancer Res 55:5217–5221, 1995

    • Search Google Scholar
    • Export Citation
  • 9.

    Harada K, , Kurisu K, & Arita K, et al: Telomerase activity in central nervous system malignant lymphoma. Cancer 86:10501055, 1999 Harada K, Kurisu K, Arita K, et al: Telomerase activity in central nervous system malignant lymphoma. Cancer 86:1050–1055, 1999

    • Search Google Scholar
    • Export Citation
  • 10.

    Harada K, , Kurisu K, & Sadatomo T, et al: Telomerase activity in human brain tumors, in International Congress of Neurological Surgery (ed): 11th International Congress of Neurological Surgery. Bologna: Monduzzi Editore, 1997, pp 17871791 Harada K, Kurisu K, Sadatomo T, et al: Telomerase activity in human brain tumors, in International Congress of Neurological Surgery (ed): 11th International Congress of Neurological Surgery. Bologna: Monduzzi Editore, 1997, pp 1787–1791

    • Search Google Scholar
    • Export Citation
  • 11.

    Harley CB, , Futcher AB, & Greider CW: Telomeres shorten during ageing of human fibroblasts. Nature 345:458460, 1990 Harley CB, Futcher AB, Greider CW: Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460, 1990

    • Search Google Scholar
    • Export Citation
  • 12.

    Harley CB, & Villeponteau B: Telomeres and telomerase in aging and cancer. Curr Opin Genet Dev 5:249255, 1995 Harley CB, Villeponteau B: Telomeres and telomerase in aging and cancer. Curr Opin Genet Dev 5:249–255, 1995

    • Search Google Scholar
    • Export Citation
  • 13.

    Hiraga S, , Ohnishi T, & Izumoto S, et al: Telomerase activity and alterations in telomere length in human brain tumors. Cancer Res 58:21172125, 1998 Hiraga S, Ohnishi T, Izumoto S, et al: Telomerase activity and alterations in telomere length in human brain tumors. Cancer Res 58:2117–2125, 1998

    • Search Google Scholar
    • Export Citation
  • 14.

    Hirose M, , Abe-Hashimoto J, & Ogura K, et al: A rapid, useful and quantative method to measure telomerase activity by hybridization protection assay connected with a telomeric repeat amplification protocol. J Cancer Res Clin Oncol 123:337344, 1997 Hirose M, Abe-Hashimoto J, Ogura K, et al: A rapid, useful and quantative method to measure telomerase activity by hybridization protection assay connected with a telomeric repeat amplification protocol. J Cancer Res Clin Oncol 123:337–344, 1997

    • Search Google Scholar
    • Export Citation
  • 15.

    Hiyama E, , Gollahon H, & Kataoka T, et al: Telomerase activity in human breast tumors. J Natl Cancer Inst 88:116122, 1996 Hiyama E, Gollahon H, Kataoka T, et al: Telomerase activity in human breast tumors. J Natl Cancer Inst 88:116–122, 1996

    • Search Google Scholar
    • Export Citation
  • 16.

    Hiyama E, , Hiyama K, & Yokoyama T, et al: Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med 1:249255, 1995 Hiyama E, Hiyama K, Yokoyama T, et al: Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med 1:249–255, 1995

    • Search Google Scholar
    • Export Citation
  • 17.

    Holt SE, & Shay JW: Role of telomerase in cellular proliferation and cancer. J Cell Physiol 180:1018, 1999 Holt SE, Shay JW: Role of telomerase in cellular proliferation and cancer. J Cell Physiol 180:10–18, 1999

    • Search Google Scholar
    • Export Citation
  • 18.

    Ishikawa F: Telomere crisis, the driving force in cancer cell evolution. Biochem Biophys Res Commun 230:16, 1997 Ishikawa F: Telomere crisis, the driving force in cancer cell evolution. Biochem Biophys Res Commun 230:1–6, 1997

    • Search Google Scholar
    • Export Citation
  • 19.

    Iwadate Y, , Fujimoto S, & Tagawa M, et al: Association of p53 gene mutation with decreased chemosensitivity in human malignant gliomas. Int J Cancer 69:236240, 1996 Iwadate Y, Fujimoto S, Tagawa M, et al: Association of p53 gene mutation with decreased chemosensitivity in human malignant gliomas. Int J Cancer 69:236–240, 1996

    • Search Google Scholar
    • Export Citation
  • 20.

    Kersemaekers AMF, , Fleuren GJ, & Kenter GG, et al: Oncogene alterations in carcinomas of the uterine cervix: overexpression of the epidermal growth factor receptor is associated with poor prognosis. Clin Cancer Res 5:577586, 1999 Kersemaekers AMF, Fleuren GJ, Kenter GG, et al: Oncogene alterations in carcinomas of the uterine cervix: overexpression of the epidermal growth factor receptor is associated with poor prognosis. Clin Cancer Res 5:577–586, 1999

    • Search Google Scholar
    • Export Citation
  • 21.

    Kim NW, , Piatyszek MA, & Prowse KR, et al: Specific association of human telomerase activity with immortal cells and cancer. Science 266:20112015, 1994 Kim NW, Piatyszek MA, Prowse KR, et al: Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015, 1994

    • Search Google Scholar
    • Export Citation
  • 22.

    Kleihues P, , Burger PC, & Scheithauer BW, et al: Histological Typing of Tumours of the Central Nervous System, ed 2. Berlin: Springer-Verlag, 1993 Kleihues P, Burger PC, Scheithauer BW, et al: Histological Typing of Tumours of the Central Nervous System, ed 2. Berlin: Springer-Verlag, 1993

    • Search Google Scholar
    • Export Citation
  • 23.

    Kleihues P, & Cavenee WK: Pathology and Genetics of Tumours of the Nervous System. Lyon: International Agency for Research on Cancer, 1997, pp 1624 Kleihues P, Cavenee WK: Pathology and Genetics of Tumours of the Nervous System. Lyon: International Agency for Research on Cancer, 1997, pp 16–24

    • Search Google Scholar
    • Export Citation
  • 24.

    Kleihues P, & Ohgaki H: Genetics of glioma progression and the definition of primary and secondary glioblastoma. Brain Pathol 7:11311136, 1997 Kleihues P, Ohgaki H: Genetics of glioma progression and the definition of primary and secondary glioblastoma. Brain Pathol 7:1131–1136, 1997

    • Search Google Scholar
    • Export Citation
  • 25.

    Kleinschmidt-DeMasters BK, , Hashizumi TL, & Sze CI, et al: Telomerase expression shows differences across multiple regions of oligodendroglioma versus high grade astrocytomas but shows correlation with Mib-1 labeling. J Clin Pathol 51:284293, 1998 Kleinschmidt-DeMasters BK, Hashizumi TL, Sze CI, et al: Telomerase expression shows differences across multiple regions of oligodendroglioma versus high grade astrocytomas but shows correlation with Mib-1 labeling. J Clin Pathol 51:284–293, 1998

    • Search Google Scholar
    • Export Citation
  • 26.

    Land H, , Parada LF, & Weinberg RA: Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304:596602, 1983 Land H, Parada LF, Weinberg RA: Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304:596–602, 1983

    • Search Google Scholar
    • Export Citation
  • 27.

    Lang FF, , Miller DC, & Koslow M, et al: Pathways leading to glioblastoma multiforme: a molecular analysis of genetic alterations in 65 astrocytic tumors. J Neurosurg 81:427436, 1994 Lang FF, Miller DC, Koslow M, et al: Pathways leading to glioblastoma multiforme: a molecular analysis of genetic alterations in 65 astrocytic tumors. J Neurosurg 81:427–436, 1994

    • Search Google Scholar
    • Export Citation
  • 28.

    Langford LA, , Piatyszek MA, & Xu R, et al: Telomerase activity in human brain tumours. Lancet 346:12671268, 1995 Langford LA, Piatyszek MA, Xu R, et al: Telomerase activity in human brain tumours. Lancet 346:1267–1268, 1995

    • Search Google Scholar
    • Export Citation
  • 29.

    Marchetti A, , Buttitta F, & Girlando S, et al: mdm2 gene alterations and mdm2 protein expression in breast carcinomas. J Pathol 175:3138, 1995 Marchetti A, Buttitta F, Girlando S, et al: mdm2 gene alterations and mdm2 protein expression in breast carcinomas. J Pathol 175:31–38, 1995

    • Search Google Scholar
    • Export Citation
  • 30.

    Meyerson M, , Counter CM, & Eaton EN, et al: hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 90:785795, 1997 Meyerson M, Counter CM, Eaton EN, et al: hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization. Cell 90:785–795, 1997

    • Search Google Scholar
    • Export Citation
  • 31.

    Nakamura TM, , Morin GB, & Chapman KB, et al: Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955959, 1997 Nakamura TM, Morin GB, Chapman KB, et al: Telomerase catalytic subunit homologs from fission yeast and human. Science 277:955–959, 1997

    • Search Google Scholar
    • Export Citation
  • 32.

    Nakayama J, , Saito M, & Nakamura H, et al: TLP1: a gene encoding a protein component of mammalian telomerase is a novel member of WD repeats family. Cell 88:875884, 1997 Nakayama J, Saito M, Nakamura H, et al: TLP1: a gene encoding a protein component of mammalian telomerase is a novel member of WD repeats family. Cell 88:875–884, 1997

    • Search Google Scholar
    • Export Citation
  • 33.

    Nakayama J, , Tahara H, & Tahara E, et al: Telomerase activation by hTRT in human normal fibroblast and hepatocellular carcinomas. Nat Genet 18:6568, 1998 Nakayama J, Tahara H, Tahara E, et al: Telomerase activation by hTRT in human normal fibroblast and hepatocellular carcinomas. Nat Genet 18:65–68, 1998

    • Search Google Scholar
    • Export Citation
  • 34.

    Reissmann PT, , Koga H, & Figlin RA, et al: Amplification and overexpression of the cyclin D1 and epidermal growth factor receptor genes in non-small-cell lung cancer. Lung Cancer Study Group. J Cancer Res Clin Oncol 125:6170, 1999 Reissmann PT, Koga H, Figlin RA, et al: Amplification and overexpression of the cyclin D1 and epidermal growth factor receptor genes in non-small-cell lung cancer. Lung Cancer Study Group. J Cancer Res Clin Oncol 125:61–70, 1999

    • Search Google Scholar
    • Export Citation
  • 35.

    Seliger B, , Papadileris S, & Vogel D, et al: Analysis of the p53 and MDM-2 gene in acute myeloid leukemia. Eur J Haematol 57:230240, 1996 Seliger B, Papadileris S, Vogel D, et al: Analysis of the p53 and MDM-2 gene in acute myeloid leukemia. Eur J Haematol 57:230–240, 1996

    • Search Google Scholar
    • Export Citation
  • 36.

    Shay JW: Telomerase in human development and cancer. J Cell Physiol 173:266270, 1997 Shay JW: Telomerase in human development and cancer. J Cell Physiol 173:266–270, 1997

    • Search Google Scholar
    • Export Citation
  • 37.

    Shay JW, , Werbin H, & Wright WE: Telomere shortening may contribute to aging and cancer: a perspective. Mol Cell Differ 2:121, 1994 Shay JW, Werbin H, Wright WE: Telomere shortening may contribute to aging and cancer: a perspective. Mol Cell Differ 2:1–21, 1994

    • Search Google Scholar
    • Export Citation
  • 38.

    Shay JW, , Wright WE, & Werbin H: Defining the molecular mechanisms of human cell immortalization. Biochim Biophys Acta 1072:17, 1991 Shay JW, Wright WE, Werbin H: Defining the molecular mechanisms of human cell immortalization. Biochim Biophys Acta 1072:1–7, 1991

    • Search Google Scholar
    • Export Citation
  • 39.

    Tahara H, , Kuniyasu H, & Yasui W, et al: Telomerase activity in gastric and colorectal carcinomas correlates well with malignant progression. Clin Cancer Res Rapid Commun 1:12451251, 1995 Tahara H, Kuniyasu H, Yasui W, et al: Telomerase activity in gastric and colorectal carcinomas correlates well with malignant progression. Clin Cancer Res Rapid Commun 1:1245–1251, 1995

    • Search Google Scholar
    • Export Citation
  • 40.

    Tahara H, , Nakanishi T, & Kitamoto M, et al: Telomerase activity in human liver tissues: comparison between chronic liver disease and hepatocellular carcinomas. Cancer Res 55:27342736, 1995 Tahara H, Nakanishi T, Kitamoto M, et al: Telomerase activity in human liver tissues: comparison between chronic liver disease and hepatocellular carcinomas. Cancer Res 55:2734–2736, 1995

    • Search Google Scholar
    • Export Citation
  • 41.

    Tahara H, , Tokutake Y, & Maeda S: Abnormal telomere dynamics of B-lymphoblastoid cell strains from Werner's syndrome patients transformed by Epstein-Barr virus. Oncogene 15:19111920, 1997 Tahara H, Tokutake Y, Maeda S: Abnormal telomere dynamics of B-lymphoblastoid cell strains from Werner's syndrome patients transformed by Epstein-Barr virus. Oncogene 15:1911–1920, 1997

    • Search Google Scholar
    • Export Citation
  • 42.

    Ullrich A, , Coussens L, & Hayflick JS, et al: Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells. Nature 309:418425, 1984 Ullrich A, Coussens L, Hayflick JS, et al: Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells. Nature 309:418–425, 1984

    • Search Google Scholar
    • Export Citation
  • 43.

    Vogelstein B, & Kinzler KW: The multistep nature of cancer. Trends Genet 9:138141, 1993 Vogelstein B, Kinzler KW: The multistep nature of cancer. Trends Genet 9:138–141, 1993

    • Search Google Scholar
    • Export Citation
  • 44.

    Watanabe K, , Sato K, & Biernat W, et al: Incidence and timing of p53 mutations during astrocytoma progression in patients with multiple biopsies. Clin Cancer Res 3:523530, 1997 Watanabe K, Sato K, Biernat W, et al: Incidence and timing of p53 mutations during astrocytoma progression in patients with multiple biopsies. Clin Cancer Res 3:523–530, 1997

    • Search Google Scholar
    • Export Citation
  • 45.

    Watanabe K, , Tachibana O, & Sato K, et al: Overexpression of the EGF receptor and p53 mutations are mutually exclusive in the evolution of primary and secondary glioblastomas. Brain Pathol 6:217224, 1996 Watanabe K, Tachibana O, Sato K, et al: Overexpression of the EGF receptor and p53 mutations are mutually exclusive in the evolution of primary and secondary glioblastomas. Brain Pathol 6:217–224, 1996

    • Search Google Scholar
    • Export Citation

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