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Signaling and Regulation

A Simple Epigenetic Method for the Diagnosis and Classification of Brain Tumors¹

¹ Department of Neurosurgery and Neurotraumatology, Karol Marcinkowski University School of Medical Sciences, Poznan, Poland; ² Faculty of Chemistry, Adam Mickiewicz University, Poznan, Poland; ³ Institute of Bioorganic Chemistry of the Polish Academy of Sciences, Poznan, Poland; and ⁴ Institut de Biologie Moléculaire et Cellulaire du Centre National de la Recherche Scientifique, Strasbourg, France

Requests for reprints: Miroslawa Z. Barciszewska, Institute of Bioorganic Chemistry of the Polish Academy of Sciences, Noskowskiego 12, 61-704 Poznan, Poland. Phone: 48-61-852-85-03; Fax: 48-61-852-05-32. E-mail: mbarcisz{at}ibch.poznan.pl

The new, simple, and reliable method for the diagnosis of brain tumors is described. It is based on a TLC quantitative determination of 5-methylcytosine (m⁵C) in relation to its damage products of DNA from tumor tissue. Currently, there is evidence that oxidative stress through reactive oxygen species (ROS) plays an important role in the etiology and progression of several human diseases. Oxidative damage of DNA, lipids, and proteins is deleterious for the cell. m⁵C, along with other basic components of DNA, is the target for ROS, which results in the appearance of new modified nucleic acid bases. If so, m⁵C residue constitutes a mutational hotspot position, whether it occurs within a nucleotide sequence of a structural gene or a regulatory region. Here, we show the results of the analysis of 82 DNA samples taken from brain tumor tissues. DNA was isolated and hydrolyzed into nucleotides, which, after labeling with [-³²P]ATP, were separated on TLC. Chromatograms were evaluated using PhosphorImager and the amounts of 5-methyldeoxycytosine (m⁵dC) were calculated as a ratio (R) of m⁵dC to m⁵dC + deoxycytosine + deoxythymidine spot intensities. The R value could not only be a good diagnostic marker for brain tumors but also a factor differentiating low-grade and high-grade gliomas. Therefore, DNA methylation pattern might be a useful tool to give a primary diagnosis of a brain tumor or as a marker for the early detection of the relapse of the disease. This method has several advantages over those existing nowadays.

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