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  • Title: Quantitative real time PCR of deoxycytidine kinase mRNA by Light Cycler PCR in relation to enzyme activity and gemcitabine sensitivity.
    Author: Sigmond J, Kroep JR, Loves W, Codacci-Pisanelli G, Peters GJ.
    Journal: Cancer Lett; 2004 Sep 30; 213(2):173-9. PubMed ID: 15327832.
    Abstract:
    Deoxycytidine kinase (dCK) is essential for the phosphorylation of gemcitabine and can predict response to gemcitabine in vivo. Conventional Competitive Template-Reverse Transcriptase-Polymerase Chain Reaction (CT-RT-PCR) was correlated with real time PCR using a Light Cycler (LC) with SYBR-Green detection to enable rapid and sensitive detection of dCK mRNA expression. We used cDNA from human xenografts to establish a relation between dCK activity and gemcitabine sensitivity. A significant correlation of LC-PCR was found with CT-RT-PCR (Pearson: r = 0.956; P < 0.0001), enzyme activity (Pearson: r = 0.972; P = 0.003) and gemcitabine sensitivity (Pearson: r = 0.695; P = 0.048). The LC-PCR was also applied to needle biopsy specimens. In bladder tumors a similar correlation was found, while esophageal tumors with a high dCK expression responded to gemcitabine treatment. The LC is a rapid and reliable method for quantitation of dCK mRNA levels in tumors to predict clinical gemcitabine sensitivity.
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