A new, simple method for quantifying gemcitabine triphosphate in cancer cells using isocratic high-performance liquid chromatography

スポンサーリンク

概要

• 論文の詳細を見る

• A deoxycytidine analog, gemcitabine (dFdC), is effective for treating solid tumors and hematologic malignancies. After being transported into cancer cells, dFdC is phosphorylated to dFdC triphosphate (dFdCTP), which is subsequently incorporated into the DNA strand, thereby inhibiting DNA synthesis. Intracellular dFdCTP is the critical determinant for dFdC cytotoxicity, so therapeutic drug monitoring or in vitro testing of the capability of cancer cells to accumulate dFdCTP may be informative for optimizing dFdC administration. We have developed a new isocratic-elution HPLC method for quantifying dFdCTP in cancer cells. Samples (500 μl) were eluted isocratically using 0.06 M Na2HPO4 (pH = 6.9) containing 20% acetonitrile, at a constant flow rate of 0.7 ml/min and at ambient temperature. Separation was performed using an anion-exchange column, TSK gel DEAE-2SW (250 mm x 4.6 mm inside diameter, particle size 5 μl, TOSOH Corp.), and monitored at 254 nm. The standard curve was linear with low within-day and inter-day variability. The lower detection limit (20 pmol) was as sensitive as that of the previous gradient-elution method. dFdCTP was well separated from other nucleoside triphosphates. The method could measure dFdCTP in cultured or primary leukemic cells treated in vitro with dFdC. The method was also applicable to simultaneous determination of dFdCTP and cytarabine triphosphate, the results of which demonstrated the ara-CTP production augmented by the dFdC pretreatment. Thus, our isocratic HPLC assay method will be of great use because of its sensitivity and simplicity as well as its applicability to biological materials.

• 2006-11-10

著者

• Yamauchi Takahiro

  Division Of Hematology And Oncology Faculty Of Medical Sciences University Of Fukui

• Ueda Takanori

  Division Of Hematology And Oncology Faculty Of Medical Sciences University Of Fukui

• NISHI Rie

  Division of Hematology and Oncology, Faculty of Medical Sciences, University of Fukui

• Nishi Rie

  Division Of Hematology And Oncology Faculty Of Medical Sciences University Of Fukui

関連論文

• Binding of monosodium urate crystals with idiotype protein efficiently promote dendritic cells to induce cytotoxic T cells
• Multiple inflammatory cytokine-productive ThyL-6 cell line established from a patient with thymic carcinoma
• Frequent detection of multidrug-resistant pneumonia-causing bacteria in the pneumonia lung tissues of patients with hematological malignancies
• A new, simple method for quantifying gemcitabine triphosphate in cancer cells using isocratic high-performance liquid chromatography
• Surveillance of the Serum Candida Antigen Titer for Initiation of Antifungal Therapy After Postremission Chemotherapy in Patients with Acute Leukemia

スポンサーリンク