Quantitative real time PCR of deoxycytidine kinase mRNA by Light Cycler PCR; in relation to enzyme activity.
Deoxycytidine kinase (dCK) is essential for the phosphorylation of several deoxyribonucleosides and various analogues such as gemcitabine (2',2'-difluorodeoxycytidine). We developed and optimized a sensitive real time Light Cycler (LC) PCR assay for dCK with SYBR green detection. The enzymatic activity measured in the same human xenografts of dCK correlated excellently with dCK mRNA expression levels measured by the LC. This assay can be used for evaluation of dCK expression in tumors.
Sigmond
J
J
Department of Medical Oncology, VU University Medical Center, Amsterdam, The Netherlands.
Loves
W J
WJ
Kroep
J R
JR
Peters
G J
GJ
D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D003842:Deoxycytidine Kinase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009368:Neoplasm Transplantation; D010766:Phosphorylation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D012313:RNA; D003842:Deoxycytidine Kinase