TY - JOUR
T1 - Methylation specific PCR to characterize methylation of the promoter of deoxycytidine kinase
AU - Peters, G. J.
AU - Hodzic, J.
AU - Ortega, B.
AU - Giovannetti, E.
AU - Adema, A. D.
AU - Broekhuizen, R.
AU - Kaspers, G. J.
AU - Hubeek, I.
PY - 2010/6
Y1 - 2010/6
N2 - Deoxycytidine kinase (dCK) is essential for phosphorylation of natural deoxynucleosides and analogs, such as gemcitabine and cytarabine, two widely used anticancer compounds. We hypothesized that DNA methylation of SP1 binding sites in the dCK promoter region might affect dCK expression. Using methylation specific PCR (MSP), methylation was detected in one of the SP1 binding sites of the dCK promoter, in most tested cancer cell lines and in patient samples from brain tumors and leukemia. This SP1 site is a 3'GC box, which upon hypomethylation negatively regulates dCK mRNA expression. In conclusion, we developed a new MSP method showing methylation of the 3' GC-box in the dCK promoter region in tumor cells and patient samples. Methylation might therefore regulate transcription of dCK, and should be studied further to understand its role in influencing gemcitabine and cytarabine activity.
AB - Deoxycytidine kinase (dCK) is essential for phosphorylation of natural deoxynucleosides and analogs, such as gemcitabine and cytarabine, two widely used anticancer compounds. We hypothesized that DNA methylation of SP1 binding sites in the dCK promoter region might affect dCK expression. Using methylation specific PCR (MSP), methylation was detected in one of the SP1 binding sites of the dCK promoter, in most tested cancer cell lines and in patient samples from brain tumors and leukemia. This SP1 site is a 3'GC box, which upon hypomethylation negatively regulates dCK mRNA expression. In conclusion, we developed a new MSP method showing methylation of the 3' GC-box in the dCK promoter region in tumor cells and patient samples. Methylation might therefore regulate transcription of dCK, and should be studied further to understand its role in influencing gemcitabine and cytarabine activity.
KW - deoxycytidine kinase
KW - Gemcitabine
KW - methylation specific PCR
KW - promoter methylation
UR - http://www.scopus.com/inward/record.url?scp=77953697072&partnerID=8YFLogxK
U2 - 10.1080/15257771003730078
DO - 10.1080/15257771003730078
M3 - Article
C2 - 20544528
AN - SCOPUS:77953697072
SN - 1525-7770
VL - 29
SP - 408
EP - 413
JO - Nucleosides, Nucleotides and Nucleic Acids
JF - Nucleosides, Nucleotides and Nucleic Acids
IS - 4-6
ER -