• Journal of Pharmaceutical Investigation
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• v.34 no.3
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• pp.161-168
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• 2004

5-Fluorouracil (5-FU) is an antimetabolite anticancer agent active against many types of solid tumors. Tegafur (TF), a prodrug of 5-FU, is frequently used in combination with uracil as dihydropyrimidine dehydrogenase (DPD) inhibitory fluoropyrimidine. We studied the stability of 5-FU and TF in biological fluids of rats and determined their bioavailability (BA) and excretion into bile, and urine. The drug concentrations were analyzed by an HPLC method. At room temperature, there was a 14-30% decrease in the concentration of 5-FU and TF in bile, urine, and plasma specimen at 10 and $100\;{\mu}g/ml$ over 240 min. No significant difference was noted among the sample types or between two different concentrations of 10 and $100{\mu}g/ml$. The decrease in drug concentration was significantly less in samples kept on ice (6-12%) for both drugs. These data indicate that biological fluid samples containing 5-FU or TF in plasma, urine, or bile should be placed on ice during the sample collection. Following these storage guidelines, samples were collected after administration 50 mg/kg of each drug via i.v. or oral route. BA was 1.5 folds greater for TF (60%) than that of 5-FU (42%). Approximately 0.52 and 3.3% of the i.v. doses of 5-FU and TF was excreted into bile, respectively. Renal clearance of 5-FU was about 16% of its total body clearance. These results suggest that instability of 5-FU and TF in biological fluids should be considered in pharmacokinetic or pharmacogenomic studies.

• The Korean Journal of Physiology and Pharmacology
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• v.16 no.1
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• pp.11-16
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• 2012

Cancer stem cells (CSCs) are often characterized by the elevated expression of drug-resistance related stem-cell surface markers, such as CD133 and ABCG2. Recently, we reported that CSCs have a high level of expression of the IL-6 receptor (IL-6R). The purpose of this study was to investigate the effect of anticancer drugs on the expression of the drug resistance-related cancer stem cell markers, ABCG2, IL-6R, and CD133 in non-small cell lung cancer (NSCLC) cell lines. A549, H460, and H23 NSCLC cell lines were treated with the anticancer drugs 5-fluorouracil (5-FU; $25{\mu}g/ml$) and methotrexate (MTX; $50{\mu}g/ml$), and the expression of putative CSC markers was analyzed by fluorescent activated cell sorter (FACS) and the gene expression level of abcg2, il-6r and cd133 by reverse transcriptase-polymerase chain reaction (RT-PCR). We found that the fraction of ABCG2-positive(+) cells was significantly increased by treatment with both 5-FU and MTX in NSCLC cells, and the elevation of abcg2, il-6r and cd133 expressions in response to these drugs was also confirmed using RT-PCR. Also, the number of IL-6R(+) cells was increased by MTX in the 3 cell lines mentioned and increased by 5-FU in the H460 cell line. The number of CD133(+) cells was also significantly increased by both 5-FU and MTX treatment in all of the cell lines tested. These results indicate that 5-FU and MTX considerably enhance the expression of drug-resistance related CSC markers in NSCLC cell lines. Thus, we suggest that antimetabolite cancer drugs, such as 5-FU and MTX, can lead to the propagation of CSCs through altering the expression of CSC markers.

• Tuberculosis and Respiratory Diseases
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• v.53 no.3
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• pp.275-284
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• 2002

Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.

• Applied Biological Chemistry
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• v.11
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• pp.151-166
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• 1969

I. Fffects of nitrogen supplying level and culture condition on the top growth aod tubers formation of Ipomoea Batatas. 1) The low level nitrogen (A plot) 3 Milliequivalent per liter of nutrient solution stimulated tuber formation while the high level nitrogen ($B_1\;and\;B_2$ plot) of 10 milliequivalent per liter failed to form tuber though fibrous roots were seen much activated. The suppressive effect of nitrogen on tuber formation in presumed to result from the direct suppressive effect of nitrogen or a certain biocatalystic effect rather than from any indirect effect through the stimulation to growth of tops or the competition with carbohydrates. 2) The addition of milligram urea to nutrient solution stimulated the growth and increased fresh weight and dry weight of the aerial part while suppressed, a little, plant length. 3) The water culture method, which this experiment newly adopted, stimulated plant growth more than the gravel Culture method. And the treatment of low level nitrogen (A plot) in this water culture also saw a considerable degree of tuber formation, as in the case of gravel culture. 4) The foliar application of growth retardant B-nine suppressed the plant length only, with no other recognizable effect. II. Fffects of urea supplying level on the growth of IPOMOEA BATATAS. 1) The higher level of urea which was absorbed tby roots through nutrient solution suppressed top growth, such as plant length, number of leaves and fresh weight. And this can be attributed to the direct absorption of urea which was not ammonificated. 2) Although the higher level of nitrate nitrogen (B plot) made no tuber formation in previous experiment (Report-1), the higher level of urea nitrogen (A plot) made tuber formation possible in this experiment. The ratio of tuber to top was, however, less in higher level of urea than in lower level of urea, and the suppressing effect was larger on tuber than on top. 3) The foliar application of urea stimulated top growth while the higher level of urea absorbed by roots suppressed it, though the amounts of urea supplied in two experiments were same. Ratio of top to roots was larger in foliar application of urea (C plot) and less in root absorption of urea both of higher (B plot) and lower urea levels (A plot). III. Fffects of growth retardant etc. on the growth of IPOMOEA BATATAS in relation to urea application. 1) B-nine (N-dimethyl amino-succinamic acid) is recognized as a growth retardant, suppressed the plant length irrespective of urea levels. The treatment of gibberellin stimulated distinctly plant length, and the combined treatment of gibberellin and B-nine recovered completely the plant length which had been suppressed by B-nine. 2) B-nine increased fresh weight, especially, fresh weight of top both in lower and higher level of The degree of fresh weight increase varied according to concentrations of B-nine, of which the 0.15% of B-nine ($B_1$ plot) was the effective in higher level of urea. The effect of B-nine for increasing fresh weight was the largest in top next in tuber, and the least in fibrous roots. The ratio of fibrous roots to top was always decreased by B-nine application, which the ratio of tuber to top was contrary increased by B-nine in higher level of urea though decreased in lower level of urea. 3) Gibberellin treatment also increased fresh weight but the combined treatment ($B_3$+GA plot) of gibberellin and B-nine was even more effective than any of single treatments. Gibberellin and B-nine proved to be synergistic with fresh weight while reverse with plant length. 4) Considerable influences were abserved mainly in the length of plants and their fresh weight after B-nine treatment. So that B-nine may be reguraded as a metabolic controller rather than as an antimetabolite. 5) The surpressed growth of plants cause by higher level of urea was normalized by B-nine treatment. This fact suggested a further study on the applicability for practical use.