Skip to main content
NCBI home page
Cellular and Molecular Neurobiology logoLink to Cellular and Molecular Neurobiology
. 2005 Sep;25(6):1067–1071. doi: 10.1007/s10571-005-8475-0

Quantitation of O6-Methylguanine-DNA Methyltransferase Gene Messenger RNA in Gliomas by Means of Real-Time RT-PCR and Clinical Response to Nitrosoureas

Satoshi Tanaka 1,✉, Hidehiro Oka 2, Kiyotaka Fujii 2, Kaoru Watanabe 3, Kumi Nagao 3, Atsushi Kakimoto 3
PMCID: PMC11529508  PMID: 16392037

Abstract

  1. O6-methylguanine-DNA methyltransferase (MGMT) mRNA was measured in 50 malignant gliomas that had received 1-(4-amino-2-methyl-5-pyrimidynyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU) after the resection of the tumor by real-time reverse transcription-polymerase chain reaction (RT-PCR) using TaqMan probe.

  2. The mean absolute value of MGMTmRNA normalized to the level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for 50 tumors was 1.29 × 104± 1.28 × 104 copy/μg RNA (mean ± SD). The amount of MGMTmRNA less than 6 × 103 copy/μg RNA was the most significant factor in predicting the initial effect of treatment with ACNU by multi-variant regression analysis (p = 0.0157).

  3. These results suggest that quantitation of MGMTmRNA is the excellent method for predicting for the effect of ACNU in glioma therapy.

Key Words: MGMT, Real-Time RT-PCR, ACNU, glioma

References

  1. Aldea, C., Alvarez, C. P., Folgueira, L., Delgado, R., and Otero, J. R. (2002). Rapid detection of herpes simplex virus DNA in genital ulcers by real-time PCR using SYBR green I dye as the detection signal. J. Clin. Microbiol.40:1060–1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arcicasa, M., Roncadin, M., Bortolus, R., Bassignano, G., Boz, G., Franchin, G., De Paoli, A., and Trovo, M. G. (1994). Results of three consecutive combined treatments for malignant gliomas. Ten-year experience at a single institution. Am. J. Clin. Oncol.17:437–443. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski, P., and Sacchi, N. (1987). Single-step methods of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159. [DOI] [PubMed] [Google Scholar]
  4. Heid, C. A., Stevens, J., Livak, K. J., and Williams, P. M. (1996). Real time quantitative PCR. Genome Res.6:986–994. [DOI] [PubMed] [Google Scholar]
  5. Mineura, K., Yanagisawa, T., Watanabe, K., Kowada, M., and Yasui, N. (1996). Human brain tumor O(6)-methylguanine-DNA methyltransferase mRNA and its significance as an indicator of selective chloroethylnitrosourea chemotherapy. Int. J. Cancer69:420–425. [DOI] [PubMed] [Google Scholar]
  6. Mullah, B., Livak, K., Andrus, A., and Kenney, P. (1998). Efficient synthesis of double dye-labeled oligodeoxyribonucleotide probes and their application in a real time PCR assay. Nucleic Acids Res. 26:1026–1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Nagane, M., Shibui, S., Oyama, H., Asai, A., Kuchino, Y., and Nomura, K. (1995). Investigation of chemoresistance-related genes mRNA expression for selecting anticancer agents in successful adjuvant chemotherapy for a case of recurrent glioblastoma. Surg. Neurol.44:462–468. [DOI] [PubMed] [Google Scholar]
  8. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A. (1988). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science239:487–491. [DOI] [PubMed] [Google Scholar]
  9. Silber, J. R., Mueller, B. A., Ewers, T. G., and Berger, M. S. (1993). Comparison of O6-methylguanine-DNA methyltransferase activity in brain tumors and adjacent normal brain. Cancer Res.53:3416–3420. [PubMed] [Google Scholar]
  10. Tanaka, S., Kamitani, H., Amin, M. R., Watanabe, T., Oka, H., Fujii, K., Nagashima, T., and Hori, T. (2000). Preliminary individual adjuvant therapy for gliomas based on the results of molecular biological analyzes for drug-resistance genes. J. Neuro-Oncol.46:157–171. [DOI] [PubMed] [Google Scholar]
  11. Tanaka, S., Kobayashi, I., Oka, H., Fujii, K., Watanabe, T., Nagashima, T., and Hori, T. (2003). O6-methylguanine-DNA methyltransferase gene expression in gliomas by quantitative real-time RT-PCR and clinical response to nitrosoureas. Int. J. Cancer103:67–72. [DOI] [PubMed] [Google Scholar]
  12. Tano, K., Shiota, S., Collier, J., Foote, R. S., and Mitra, S. (1990). Isolation and structural characterization of a cDNA clone encoding the human DNA repair protein for O6-alkylguanine. Proc. Natl. Acad. Sci. U.S.A.87:686–690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Yoshida, J., Kajita, Y., Wakabayashi, T., and Sugita, K. (1994). Long-term follow up results of 175 patients with malignant glioma: Importance of radical tumor resection and postoperative adjuvant therapy with interferon, ACNU and radiation. Acta. Neurochir. (Wien) 127:55–59. [DOI] [PubMed] [Google Scholar]

Articles from Cellular and Molecular Neurobiology are provided here courtesy of Springer

RESOURCES