Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma

Eiso Hiyama, Keiko Hiyama, Takashi Yokoyama, Ikuko Fukuba, Hiroaki Yamaoka, Jerry W. Shay, Yuichiro Matsuura

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Amplification of the MYCN gene and high telomerase activity predict a poor prognosis for the patients with neuroblastoma. We used PCR techniques for rapid detection of MYCN gene amplification and human telomerase reverse transcriptase (hTERT) expression in neuroblastoma specimens. The detection of MYCN gene amplification is based on differential PCR in which three primer pairs were used to coamplify a 178-bp fragment of target MYCN gene with two reference gene fragments, a 237-bp of p53 exon 7 and a 120-bp of β-globin exon 3, in a single tube of 40 surgically resected tumor samples. MYCN amplification was identified by this differential PCR in all 10 samples carrying more than 10 copies (already known to have MYCN gene amplification by Southern blot analysis). There were no false-negative or false-positive cases, and the relative intensity of MYCN bands in the differential PCR correlated significantly with the copy number determined by Southern blot analysis (γ = 0.99, P < 0.0001). This protocol was also applicable in the biopsy or aspirated samples, as well as the paraffin-embedded tissues, and in detecting intratumoral heterogeneity. Using RT-PCR procedures, hTERT mRNA expression was detectable in all 13 tumors with high telomerase activity. These nonradioisotopic PCR-based protocols for detecting MYCN gene amplification and hTERT mRNA expression are rapid and reliable and are likely to be useful to determine the biological behavior of neuroblastoma.

Original languageEnglish (US)
Pages (from-to)601-609
Number of pages9
JournalClinical Cancer Research
Volume5
Issue number3
StatePublished - Mar 1999

Fingerprint

Gene Amplification
Telomerase
Neuroblastoma
Polymerase Chain Reaction
Southern Blotting
Exons
Messenger RNA
Globins
Paraffin
Genes
Neoplasms
Biopsy
human TERT protein

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Hiyama, E., Hiyama, K., Yokoyama, T., Fukuba, I., Yamaoka, H., Shay, J. W., & Matsuura, Y. (1999). Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma. Clinical Cancer Research, 5(3), 601-609.

Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma. / Hiyama, Eiso; Hiyama, Keiko; Yokoyama, Takashi; Fukuba, Ikuko; Yamaoka, Hiroaki; Shay, Jerry W.; Matsuura, Yuichiro.

In: Clinical Cancer Research, Vol. 5, No. 3, 03.1999, p. 601-609.

Research output: Contribution to journalArticle

Hiyama, E, Hiyama, K, Yokoyama, T, Fukuba, I, Yamaoka, H, Shay, JW & Matsuura, Y 1999, 'Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma', Clinical Cancer Research, vol. 5, no. 3, pp. 601-609.
Hiyama E, Hiyama K, Yokoyama T, Fukuba I, Yamaoka H, Shay JW et al. Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma. Clinical Cancer Research. 1999 Mar;5(3):601-609.
Hiyama, Eiso ; Hiyama, Keiko ; Yokoyama, Takashi ; Fukuba, Ikuko ; Yamaoka, Hiroaki ; Shay, Jerry W. ; Matsuura, Yuichiro. / Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma. In: Clinical Cancer Research. 1999 ; Vol. 5, No. 3. pp. 601-609.
@article{4a8ddfb6d56441548891ab799fea8b10,
title = "Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma",
abstract = "Amplification of the MYCN gene and high telomerase activity predict a poor prognosis for the patients with neuroblastoma. We used PCR techniques for rapid detection of MYCN gene amplification and human telomerase reverse transcriptase (hTERT) expression in neuroblastoma specimens. The detection of MYCN gene amplification is based on differential PCR in which three primer pairs were used to coamplify a 178-bp fragment of target MYCN gene with two reference gene fragments, a 237-bp of p53 exon 7 and a 120-bp of β-globin exon 3, in a single tube of 40 surgically resected tumor samples. MYCN amplification was identified by this differential PCR in all 10 samples carrying more than 10 copies (already known to have MYCN gene amplification by Southern blot analysis). There were no false-negative or false-positive cases, and the relative intensity of MYCN bands in the differential PCR correlated significantly with the copy number determined by Southern blot analysis (γ = 0.99, P < 0.0001). This protocol was also applicable in the biopsy or aspirated samples, as well as the paraffin-embedded tissues, and in detecting intratumoral heterogeneity. Using RT-PCR procedures, hTERT mRNA expression was detectable in all 13 tumors with high telomerase activity. These nonradioisotopic PCR-based protocols for detecting MYCN gene amplification and hTERT mRNA expression are rapid and reliable and are likely to be useful to determine the biological behavior of neuroblastoma.",
author = "Eiso Hiyama and Keiko Hiyama and Takashi Yokoyama and Ikuko Fukuba and Hiroaki Yamaoka and Shay, {Jerry W.} and Yuichiro Matsuura",
year = "1999",
month = "3",
language = "English (US)",
volume = "5",
pages = "601--609",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "3",

}

TY - JOUR

T1 - Rapid detection of MYCN gene amplification and telomerase expression in neuroblastoma

AU - Hiyama, Eiso

AU - Hiyama, Keiko

AU - Yokoyama, Takashi

AU - Fukuba, Ikuko

AU - Yamaoka, Hiroaki

AU - Shay, Jerry W.

AU - Matsuura, Yuichiro

PY - 1999/3

Y1 - 1999/3

N2 - Amplification of the MYCN gene and high telomerase activity predict a poor prognosis for the patients with neuroblastoma. We used PCR techniques for rapid detection of MYCN gene amplification and human telomerase reverse transcriptase (hTERT) expression in neuroblastoma specimens. The detection of MYCN gene amplification is based on differential PCR in which three primer pairs were used to coamplify a 178-bp fragment of target MYCN gene with two reference gene fragments, a 237-bp of p53 exon 7 and a 120-bp of β-globin exon 3, in a single tube of 40 surgically resected tumor samples. MYCN amplification was identified by this differential PCR in all 10 samples carrying more than 10 copies (already known to have MYCN gene amplification by Southern blot analysis). There were no false-negative or false-positive cases, and the relative intensity of MYCN bands in the differential PCR correlated significantly with the copy number determined by Southern blot analysis (γ = 0.99, P < 0.0001). This protocol was also applicable in the biopsy or aspirated samples, as well as the paraffin-embedded tissues, and in detecting intratumoral heterogeneity. Using RT-PCR procedures, hTERT mRNA expression was detectable in all 13 tumors with high telomerase activity. These nonradioisotopic PCR-based protocols for detecting MYCN gene amplification and hTERT mRNA expression are rapid and reliable and are likely to be useful to determine the biological behavior of neuroblastoma.

AB - Amplification of the MYCN gene and high telomerase activity predict a poor prognosis for the patients with neuroblastoma. We used PCR techniques for rapid detection of MYCN gene amplification and human telomerase reverse transcriptase (hTERT) expression in neuroblastoma specimens. The detection of MYCN gene amplification is based on differential PCR in which three primer pairs were used to coamplify a 178-bp fragment of target MYCN gene with two reference gene fragments, a 237-bp of p53 exon 7 and a 120-bp of β-globin exon 3, in a single tube of 40 surgically resected tumor samples. MYCN amplification was identified by this differential PCR in all 10 samples carrying more than 10 copies (already known to have MYCN gene amplification by Southern blot analysis). There were no false-negative or false-positive cases, and the relative intensity of MYCN bands in the differential PCR correlated significantly with the copy number determined by Southern blot analysis (γ = 0.99, P < 0.0001). This protocol was also applicable in the biopsy or aspirated samples, as well as the paraffin-embedded tissues, and in detecting intratumoral heterogeneity. Using RT-PCR procedures, hTERT mRNA expression was detectable in all 13 tumors with high telomerase activity. These nonradioisotopic PCR-based protocols for detecting MYCN gene amplification and hTERT mRNA expression are rapid and reliable and are likely to be useful to determine the biological behavior of neuroblastoma.

UR - http://www.scopus.com/inward/record.url?scp=0033037407&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033037407&partnerID=8YFLogxK

M3 - Article

C2 - 10100712

AN - SCOPUS:0033037407

VL - 5

SP - 601

EP - 609

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 3

ER -