Doxifluridine is a pyrimidine derivative and is activated to 5-fluorouracil by pyrimidine phosphorylase. Multiple acral hyperpigmented macules have been reported in patients treated with systemic 5-fluorouracil or some of its prodrugs. However, there have been no reports of this adverse event being induced by doxifluridine. Herein we present a 42-year-old woman with multiple pigmented lesions on the palm and sole after chemotherapy with oral doxifluridine.
Adult ; Female ; Floxuridine ; Fluorouracil ; Humans ; Hyperpigmentation ; Immunosuppressive Agents ; Prodrugs ; Pyrimidines

OBJECTIVETo investigate the inhibiting impact of 5'-deoxy-5-fluorouridine (5'-DFUR) on human colon carcinoma cell line LOVO after transfection of thymidine phosphorylase (TP) cDNA.

METHODSTP cDNA was transfected into human colon carcinoma cell line LOVO with lentiviral vector pLenti6.3_MCS_IRES2-EGFP, and the transfection efficiency was analyzed by flow cytometry. TP mRNA and protein expressions were detected by RT-PCR and Western blotting respectively. The IC50 of 5'-DFUR on TP-transfected LOVO and parental cell were evaluated by MTT assay. The volumes of 5-FU converted from 5'-DFUR in media, where TP-transfected and parental LOVO were cultured, were detected by HPLC.

RESULTSThe stable transfectants passed 5 generations were obtained and the transfection rate was 95%. Compared with parental cell, the RQ values of mRNA expression in TP-transfected LOVO was (282.5±86.8) folds higher significantly (P<0.01), also the TP protein expression of TP-transfected LOVO was obviously up-regulated as compared to parental cells. The IC50 value of 5'-DFUR of TP-transfectants was (1087.7±89.1) μmol/L, less than (1607.3±56.8) μmol/L of parental cells significantly (P<0.01), while there was no significant difference between parental cells and vector-transfectants [(1699.5±38.7) μmol/L, P>0.05]. HPLC revealed that when medium was added with 0, 500, 1000, and 2000 μmol/L of 5'-DFUR respectively, 0, 2.10, 3.13, and 7.19 μmol/L of 5-FU was found in the parental cells culture, while 0, 22.16, 30.94 and 40.02 μmol/L of 5-FU was found in TP-transfectants culture, but no 5-FU was found in the vector-transfectants culture.

CONCLUSIONTP cDNA transfection into LOVO can up-regulate the TP mRNA and protein expressions, increase the 5-FU converted from 5'-DFUR, and enhance the cytotoxic effect of 5'-DFUR on the LOVO cells.

Cell Line, Tumor ; Colonic Neoplasms ; pathology ; DNA, Complementary ; genetics ; Floxuridine ; pharmacology ; Humans ; Thymidine Phosphorylase ; genetics ; Transfection

Several experimental and clinical studies have shown that leucovorin(LV) potentiate the antitumor effect of 5-fluorouracil(SFU) by biochemical modulation. LV increases the intracellular reduced folate, which stabilizes the 5FU metabolite 5-fluorodeoxyuridine monophosphate (FdUMP):thymidylate synthase(TS) complex, thus increasing the block in the DNA synthesis pathway. This present study was designed to analyze the cytotoxic effects of the combination of 5FU and LV in bladder cancer cell lines. We used two human bladder transitional cell cancer cell lines (DU4184 and DU4284): In vitro cytotoxicity was assessed by the colorimetric assay. Them was a substantial difference of 5FU cytotoxicity between two cell lines. DU4284 cells were 2-fold less sensitive to 5FU than DU4184 cells (mean IC50 : 0.43M+0.02 vs. 0.21+0.0l.uM). Enhanced cytotoxic effect of 5FU by coadministration of LV was seen in both cell lines with the greater degree of potentiation being observed in DU4284 cells (enhancement factor: 7.2 vs. 2.7 fold, respectively). It is concluded that, in this study, 1) bladder cancer cells reveal various patterns of cytotoxic responses to 5FU only, 2) LV significantly enhances the cytotoxicity of 5FU in bladder cancer cells including the 5FU- less sensitive cells. These data support the use of LV in combination with 5FU as adjuvant treatment of patients with advanced metastatic bladder cancer.
Cell Line* ; DNA ; Floxuridine ; Fluorouracil ; Folic Acid ; Humans* ; Inhibitory Concentration 50 ; Leucovorin* ; Urinary Bladder Neoplasms* ; Urinary Bladder*

Several experimental and clinical studies have shown that leucovorin(LV) potentiate the antitumor effect of 5-fluorouracil(SFU) by biochemical modulation. LV increases the intracellular reduced folate, which stabilizes the 5FU metabolite 5-fluorodeoxyuridine monophosphate (FdUMP):thymidylate synthase(TS) complex, thus increasing the block in the DNA synthesis pathway. This present study was designed to analyze the cytotoxic effects of the combination of 5FU and LV in bladder cancer cell lines. We used two human bladder transitional cell cancer cell lines (DU4184 and DU4284): In vitro cytotoxicity was assessed by the colorimetric assay. Them was a substantial difference of 5FU cytotoxicity between two cell lines. DU4284 cells were 2-fold less sensitive to 5FU than DU4184 cells (mean IC50 : 0.43M+0.02 vs. 0.21+0.0l.uM). Enhanced cytotoxic effect of 5FU by coadministration of LV was seen in both cell lines with the greater degree of potentiation being observed in DU4284 cells (enhancement factor: 7.2 vs. 2.7 fold, respectively). It is concluded that, in this study, 1) bladder cancer cells reveal various patterns of cytotoxic responses to 5FU only, 2) LV significantly enhances the cytotoxicity of 5FU in bladder cancer cells including the 5FU- less sensitive cells. These data support the use of LV in combination with 5FU as adjuvant treatment of patients with advanced metastatic bladder cancer.
Cell Line* ; DNA ; Floxuridine ; Fluorouracil ; Folic Acid ; Humans* ; Inhibitory Concentration 50 ; Leucovorin* ; Urinary Bladder Neoplasms* ; Urinary Bladder*

For the study on the teratogenic effects and its mechanism of FUDR on the developing rat fetuses, Sprague -Dawley rats as experimental animal and 5 -fluoro -2 '-deoxyuridine (Sigma chemicals) as FUDR were used respectively. On the day of gestation 10.5, 60 mg/kg, 65 mg/kg and 70 mg/kg of FUDR was injected intraperitoneally, and sacrificed under ether anesthesia on the day of gestation 17.5. External congenital malformations such as hydrocephalus, lens defect, cleft palate, short tail, forelimb and hindlimb malformations were observed under stereoscope, and compared each other with control group. The results were as follows; 1. Congenital anomalies induced by FUDR were hydrocephalus, lens defect, cleft palate, short tail, and defects of extremities such as phocomelia, loss of first and second digits, undergrowth of digits, and syndactyly. 2. There is close relation between the rate of occurance of congenital anomalies and doses of FUDR in some organs. 3. There is significant differences between forelimbs and hindlimbs and between right forelimb and left forelimb in the rate of occurance of congenital anomalies of the extremities. According to the above results, it is considered that there is significant differences between the types and occurance rates of congenital anomalies induced by FUDR and the doses of FUDR. But distinct mechanism of action during teratogenesis of FUDR still remains unclear.
Anesthesia ; Animals ; Cleft Palate ; Ectromelia ; Ether ; Extremities ; Fetus ; Floxuridine* ; Forelimb ; Hindlimb ; Hydrocephalus ; Pregnancy ; Rats* ; Syndactyly ; Tail ; Teratogenesis

OBJECTIVESTo study the change of ability to transform from 5'-deoxy-fluorouracil monophosphate (5'-DFUR) to fluorouracil (5-FU) in human colon cancer cell lines SW480 and LOVO which transfected with thymidine phosphorylase (TP) gene. And to discuss the anti-cancer activity of 5'-DFUR to SW480 and LOVO cells.

METHODSTP cDNA were transfected into human colorectal cancer cell lines SW480 and LOVO with the lentiviral vector, pLenti6.3_MCS_IRES2-EGFP. The transfection efficiency was analyzed by flow cytometer, the mRNA expression of TP was detected by RT-PCR, and the TP protein expression was detected by Western blot, and the volumes of 5-FU converted from 5'-DFUR both in 2 cells and medium were detected by high performance liquid chromatography (HPLC). The 50% inhibitory concentration (IC50) of 5'-DFUR on these 2 colon cancer cell lines both wild type and TP-transfected cells were evaluated by MTT assay.

RESULTSThe colorectal cancer cell lines SW480 and LOVO transfected with human TP cDNA were monitored 5 generations, and the transfections efficiency rate wea about 95%. Compared with wild type cell SW480 and LOVO, the RQ values of mRNA expression of SW480-TP and LOVO-TP were (695 ± 171) folds (t = -7.00, P = 0.002) and (282 ± 87) folds (t = -5.61, P = 0.030), respectively. Also TP protein expression in SW480-TP and LOVO-TP were higher than their parent cells shown by Western blot. The volume of 5-FU converted from 5'-DFUR in the medium cultured SW480-TP and LOVO-TP were increased compared with their parent cells, respectively (t = 19.406-66.921, P < 0.01), whereas few of 5-FU was detected both in wild, and TP-transfected cells. After transfected with TP cDNA, the IC50 of 5'-DFUR on SW480-TP and LOVO-TP were (587 ± 17) µmol/L and (1088 ± 89) µmol/L respectively, and there were significantly less than their parent cells (t = -32.59 and -8.52, P < 0.01).

CONCLUSIONSThe stabilized transfections of SW480 and LOVO with higher TP expression could be built with lentiviral vector. Transfected TP cDNA into SW480 and LOVO, could improve the expression both of TP mRNA and TP protein, increase the volume of 5-FU converted from 5'-DFUR in medium, and result in an enhancement of anticancer effect on these 2 cells.

Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Floxuridine ; metabolism ; Fluorouracil ; metabolism ; Humans ; Thymidine Phosphorylase ; genetics ; Transcription, Genetic ; Transfection

OBJECTIVETo detect the effect of interferon-α2a(IFN-α2a) on thymidine phosphorylase(TP) mRNA expression levels and the anticancer activity of 5-fluorouracil(5-FU) and 5'-deoxy-fluorouridine(5'-DFUR) in human colon carcinoma cell lines LOVO and SW480.

METHODSTwo human colon cancer cell lines LOVO and SW480 were cultured and treated with IFN-α2a at a series of dosage, and fluorescence quantitative PCR was carried out to detect the TP mRNA expression levels in these 2 cell lines. Then MTT assay and software Templet were used to determine the change of 50% inhibition concentration of 5-FU or 5'-DFUR combined with IFN-α2a on the two cell lines.

RESULTSThe TP mRNA expressions were up-regulated significantly by IFN-α2a at the doses of 500 U/ml and 5000 U/ml in LOVO(P<0.01). Compared with untreated cells(IFN-α2a 0 U/ml), no significance was found for TP mRNA expression levels in LOVO and SW480 treated by IFN-α2a at the dose of 50 U/ml (P>0.05). There was no significant difference for TP mRNA expression in SW480 between the dose of 0 U/ml and 500 U/ml of IFN-α2a(P>0.05), while a significant increace was detected at the dose of 5000 U/ml (P<0.01). No significant difference was found for the IC50 values after treatment of 5-FU combined with IFN-α2a (20 U/ml) on LOVO and SW480 compared with 5-FU alone, while the IC50 values after treatment of 5'-DFUR combined with IFN-α2a decreased significantly compared with 5'-DFUR alone(P<0.05).

CONCLUSIONThere is no direct inhibition effect of IFN-α2a on LOVO and SW480 in vitro, while it can up-regulate TP mRNA expression levels both in LOVO and SW480, and enhance the anticancer effect of 5'-DFUR on these 2 cell lines.

Cell Line, Tumor ; Colonic Neoplasms ; enzymology ; pathology ; Floxuridine ; pharmacology ; Humans ; Interferon-alpha ; pharmacology ; Recombinant Proteins ; pharmacology ; Thymidine Phosphorylase ; metabolism

A 45-year-old man was found to have advanced cancer of the gastric antrum and lower body with multiple liver metastases. A palliative subtotal gastrectomy was performed, and multiple hepatic lesions were treated by hepatic arterial infusion therapy with floxuridine (FUdR) 3 weeks after the operation. This therapy was given for 14 days every 3 weeks. He received 4 cycles of the therapy. A systemic combination of chemotherapy with 5-FU and cisplatin was also perfomed. These two courses of intraarterial infusion therapy produced marked regression of liver metastases and necrosis. The effect was, thus, rated as a partial response. However, after the 4th course of the therapy, the patient dropped out for personal reasons. A brain metastasis was found 4 months later, and this intraarterial infusion therapy could no longer be performed. This case indicates that intraarterial infusion chemotherapy with FUdR may be useful in treating multiple liver metastases from gastric cancer.
Brain ; Cisplatin ; Drug Therapy ; Floxuridine* ; Fluorouracil ; Gastrectomy ; Humans ; Infusions, Intra-Arterial ; Liver* ; Middle Aged ; Necrosis ; Neoplasm Metastasis ; Pyloric Antrum ; Stomach Neoplasms*

Exogenous factors, including environmental substances and drugs, are known to induce scleroderma-like reactions. Various scleroderma-like reactions induced by anti-cancer drugs have recently been reported. This is the first report that doxifluridine (Didox), an oral prodrug of the antineoplastic agent 5-fluorouracil (5-FU), induced a localized sclerderma-like reaction. A 51-year-old woman was referred to our clinic with multiple pearly, shiny patches on both her breasts, her pelvis and her back. After surgical excision and radiation therapy due to her right breast cancer, she took Didox for 7 months. A skin biopsy specimen revealed that the dermal collagen thickening extended even to the subcutaneous tissue. The routine laboratory tests were within the normal ranges and the tests for antinuclear antibody (ANA), anti SS-A antibody, anti SS-B antibody and anti U1-RNP antibody were all negative. After discontinuation of Didox, the lesions gradually improved. Based on these finding, we diagnosed this case as a localized scleroderma-like reaction induced by doxifluridine and we should pay attention to detect this adverse effect of the long term use of doxifluridine.
Antibodies, Antinuclear ; Biopsy ; Breast ; Breast Neoplasms ; Collagen ; Female ; Floxuridine ; Fluorouracil ; Humans ; Hydroxamic Acids ; Middle Aged ; Pelvis ; Reference Values ; Skin ; Subcutaneous Tissue

OBJECTIVEPrimary to search the rule of pre-operative chemotherapy and suitable duration for it by investigating the changes of calpain content and activity after 5'-deoxy-5-fluorouridine (5'-DFUR) per oral administered pre-operatively in different time. Further to investigate the mechanism of chemotherapy.

METHODSSeventy-three patients with gastrointestinal malignant tumors were divided into 4 groups by the time of 5'-DFUR (600 approximately 1200 mg/d) by oral administration before operation, group A, 3 days, 27 cases; group B, 1 week, 22 cases; group C, 2 weeks, 15 cases; group D, 2 months, 9 cases. And group E, control group, had 24 patients with gastrointestinal malignant tumors at the same term. The patients above all had not received the other chemotherapy and radiotherapy. Western blot and immunoelectron microscopy were employed to detect the expressing levels and activities of calpain in tumor tissues of different groups.

RESULTSElectronic microscopic examination showed gold granula mainly on the membranes of mitochondria of tumor cells to groups after chemotherapy. And the tumor cells of group A were mildly damaged. Besides that, serious injury for tumor cells of group B could be seen, and the phenomena were common in group C. But the damages to tumor cells of group D were mainly about mildness. The results of immunoelectron microscopy revealed that the contents of calpain increased following the time of chemotherapy prolonging, and peaked in group C. Still more, there was no significant difference for the results between group C and group D. The changes of calpain activities observed by western blot had the same tendency as the results from immunoelectron microscopy (r = 0.86, P < 0.0001).

CONCLUSIONS5'-DFUR via oral administered pre-operation could have anti-cancer effect through calpain. And the effect might be strongest in 2 weeks also after chemotherapy.

Administration, Oral ; Adult ; Aged ; Antimetabolites, Antineoplastic ; therapeutic use ; Calpain ; metabolism ; Chemotherapy, Adjuvant ; Female ; Floxuridine ; therapeutic use ; Gastrointestinal Neoplasms ; drug therapy ; enzymology ; ultrastructure ; Humans ; Male ; Middle Aged ; Time Factors