Development and application of a multiplex PCR procedure for the detection of DNA methylation in colorectal cancer

  • Authors:
    • Charles Anacleto
    • Benedito Rossi
    • Ademar Lopes
    • Fernando A. Soares
    • José Cláudio C. Rocha
    • Otávia Caballero
    • Anamaria A. Camargo
    • Andrew J.G. Simpson
    • Sérgio D.J. Pena
  • View Affiliations

  • Published online on: February 1, 2005     https://doi.org/10.3892/or.13.2.325
  • Pages: 325-328
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Abstract

Cancers often exhibit aberrant methylation of gene promoter regions associated with loss of tumor suppressor and/or DNA repair gene function. Such methylation constitutes an excellent marker for the molecular detection of micro-metastases and the diagnosis of tumor recurrences. We have developed a multiplex methylation-specific PCR (MSP) procedure for rapid and simultaneous assessment of the methylation of 5 loci: the tumor suppressor genes p16INK4a, death-associated protein kinase (DAPK) and p14ARF, and the DNA repair genes hMLH1 and O6-methylguanine-DNA-methyltransferase (MGMT). This multiplex test uses one single PCR reaction and only one electrophoretic run. In 98 samples of colorectal cancer studied, methylation of MGMT, DAPK, p16, hMLH1 and p14 was present in 31, 20, 17, 16 and 14% of tumors, respectively. In 58% of the tumors at least one methylated gene was found. This multiplex MSP constitutes a simple and inexpensive method for screening of molecular signatures in colorectal cancer and can be used profitably before employing more expensive and complex techniques such as microarray testing.

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February 2005
Volume 13 Issue 2

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Anacleto C, Rossi B, Lopes A, Soares FA, Rocha JC, Caballero O, Camargo AA, Simpson AJ and Pena SD: Development and application of a multiplex PCR procedure for the detection of DNA methylation in colorectal cancer. Oncol Rep 13: 325-328, 2005
APA
Anacleto, C., Rossi, B., Lopes, A., Soares, F.A., Rocha, J.C., Caballero, O. ... Pena, S.D. (2005). Development and application of a multiplex PCR procedure for the detection of DNA methylation in colorectal cancer. Oncology Reports, 13, 325-328. https://doi.org/10.3892/or.13.2.325
MLA
Anacleto, C., Rossi, B., Lopes, A., Soares, F. A., Rocha, J. C., Caballero, O., Camargo, A. A., Simpson, A. J., Pena, S. D."Development and application of a multiplex PCR procedure for the detection of DNA methylation in colorectal cancer". Oncology Reports 13.2 (2005): 325-328.
Chicago
Anacleto, C., Rossi, B., Lopes, A., Soares, F. A., Rocha, J. C., Caballero, O., Camargo, A. A., Simpson, A. J., Pena, S. D."Development and application of a multiplex PCR procedure for the detection of DNA methylation in colorectal cancer". Oncology Reports 13, no. 2 (2005): 325-328. https://doi.org/10.3892/or.13.2.325