Enhancement of the Anti‐tumor Effect of 5′‐Deoxy‐5‐fluorouridine by Transfection of Thymidine Phosphorylase Gene into Human Colon Cancer Cells

Kanyama, Hiroki; Tomita, Naohiro; Yamano, Tomoki; Miyoshi, Yasuo; Ohue, Masayuki; Fujiwara, Yoshiyuki; Sekimoto, Mitsugu; Sakita, Isao; Tamaki, Yasuhiro; Monden, Morito

doi:10.1111/j.1349-7006.1999.tb00769.x

Cited by 15 publications

(11 citation statements)

References 21 publications

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“…Correlations between TP expression level and, on one hand the in vitro fluoropyrimidine cytotoxicity (Schwartz et al, 1995;Evrard et al, 1999aEvrard et al, , 1999b, on the other hand the clinical outcome of fluoropyrimidine-based therapy (Koizumi et al, 1999;Saito et al, 1999) have been established. Thus, TP was proposed as a potential suicide gene for cancer gene therapy with 5′DFUR (Haraguchi et al, 1993;Patterson et al, 1995;Kanyama et al, 1999) or with 5-FU (Evrard et al, 1999a(Evrard et al, , 1999b (FUrd, FUMP, FUDP and FUTP). These data were in agreement with previous reports (Kufe and Major, 1981;Peter et al, 1986;Patterson et al, 1995;Ackland and Peters, 1999) showing that MCF-7 cells can activate 5-FU through both RNA (initiated by TP) and DNA (initiated by FU conversion to FUMP by orotate phosphoribosyl transferase) pathways.…”

Section: Discussionmentioning

confidence: 99%

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

Cuq¹,

Rouquet²,

Evrard³

et al. 2001

Br J Cancer

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The pyrimidine nucleoside phosphorylases (PyNP), uridine (UP; EC 2.4.2.3) and thymidine (TP; EC 2.4.2.4) phosphorylase, are involved in the salvage pathway of pyrimidine nucleosides, but also in the activation of fluoropyrimidines. They are responsible for the conversion of the pro-drug 5′-deoxy-5-fluorouridine (5′-DFUR, doxifluridine) to its active form 5-fluorouracil (5-FU) (Armstrong and Diasio, 1980). Because of their reversible phosphorolytic activity, UP and TP also assume the conversion of 5-FU to its anabolites, respectively 5-fluorouridine (5-FUrd) and 5-fluoro-2′-deoxyuridine (5-FdUrd) (Ishitsuka et al, 1980;Peters et al, 1986).PyNP gene transfer into cancer cells in order to increase the metabolic activation of fluoropyrimidines and thus the sensitivity of transfected cells to these anti-metabolites could be a new strategy in cancer gene therapy. Encouraging data have been obtained with TP gene transfer (Patterson et al, 1995;Schwartz et al, 1995;Kato et al, 1997;Evrard et al, 1999aEvrard et al, , 1999b. Unlike TP, the data concerning UP are not numerous. NIH-3T3-derived cells expressing high levels of UP are more sensitive to fluoropyrimidines than parental cells . The increase of 5′DFUR antitumor activity by an extract from calf thymus (thymosin fraction 5) was attributed to an increase of UP activity (Ikemoto et al, 1999). In this paper, we studied the effects of UP overexpression by gene transfer in human breast cancer cells on the metabolic activation and on the cytotoxicity of fluoropyrimidines. MATERIALS AND METHODS Chemicals Cell linesMCF-7 human breast adenocarcinoma cells (ATCC number HTB-22) were cultured at 37˚C in a fully humidified 5% CO 2 atmosphere in RPMI 1640 medium supplemented with 10% fetal calf serum (FCS) and 2 mM glutamine. COS-7 cells (ATCC number CRL-1651) were cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 5% FCS and 2 mM glutamine. UP overexpression in COS-7 and MCF-7 cellsThe pUP-SG5 plasmid, containing the full length human UP cDNA (Watanabe and Uchida, 1995), was kindly provided by Dr SH Watanabe (Nippon Roche Research Center, Japan). The labelling of UP cDNA by a short myc sequence was performed by PCR using pUP-SG5, pFU polymerase (Stratagene, Amsterdam, The Netherlands) and specific primers. The myc/UP cDNA was ligated into the mammalian expression vector pcDNA3 (Invitrogen, NV Leek, The Netherlands) to produce pcmyc/UP. Cells were transfected with pcmyc/UP, pUP-SG5 or pcDNA3 (control) using Lipofectamine™ (Life Technologies, CergyPontoise, France). COS-7 cells were lysed after 48 h incubation. Stable MCF-7 transfectants were selected for 250 µg ml -1 geneticin (Life Technologies, Cergy-Pontoise, France) resistance. ImmunoblottingProteins from cell lysates were electrophoresed on SDS-polyaclamide gel, electroblotted onto nitrocellulose membrane, labelled with an anti-myc antibody (Invitrogen, France) and visualized using X-ray films after incubation with ECL reagents as previously described (Evrard et al, 1999b). Summary The relationship between uri...

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Section: Discussionmentioning

confidence: 99%

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

Cuq¹,

Rouquet²,

Evrard³

et al. 2001

Br J Cancer

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“…injection of 5′-DFUR compared with that in control mice that received phosphate-buffered saline (PBS) treatment. 55) A role for TP in regulating 5′-DFUR conversion to 5-FU in tumors has also been implied, based on a study of the interplay between TP and dihydropyrimidine dehydrogenase (DPD), an enzyme that degrades 5-FU to inactive molecules. Measurement of the TP/ DPD ratio in tumor tissue obtained from surgically resected samples from 93 patients with primary gastric cancer showed a significant correlation with 5′-DFUR sensitivity.…”

Section: Correlation Between Tp Activity and Sensitivity To Chemotherapymentioning

confidence: 99%

“…53) Transfection of TP cDNA into other cultured cell lines, such as MCF-7, C26 and SW480, also enhanced their sensitivity to the prodrug 5′-DFUR. 52,54,55) The in vivo growth of the s.c. transplanted SW480/TP tumor in nude mice was significantly suppressed by i.p. injection of 5′-DFUR compared with that in control mice that received phosphate-buffered saline (PBS) treatment.…”

Section: Correlation Between Tp Activity and Sensitivity To Chemotherapymentioning

confidence: 99%

The role of thymidine phosphorylase, an angiogenic enzyme, in tumor progression

et al. 2004

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“…29 This is of interest given the relationship between colon cancer drug resistance and invasive/metastatic ability. 30 The nanomedicine inhibited cell proliferation in a dose-dependent manner with a much lower IC 50 (concentrations of the drug needed to reduce population growth by 50% in vitro) value (0.1 µM) than that reported previously for SW480 cells (IC 50 5-FU: 4 µM), 31 showing that the antitumor effect of 5-FU-loaded PCL NPs in SW480 was 40-fold higher than that for the free drug ( Figure 3B). …”

Section: Antitumor Activity Of 5-fu-loaded Pcl Nps On Sw480 Cellsmentioning

confidence: 72%

5-Fluorouracil-loaded poly(ε-caprolactone) nanoparticles combined with phage E gene therapy as a new strategy against colon cancer

Ortíz¹,

Prados²,

Melguizo³

et al. 2012

IJN

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This work aimed to develop a new therapeutic approach to increase the efficacy of 5-fluorouracil (5-FU) in the treatment of advanced or recurrent colon cancer. 5-FU-loaded biodegradable poly(ε-caprolactone) nanoparticles (PCL NPs) were combined with the cytotoxic suicide gene E (combined therapy). The SW480 human cancer cell line was used to assay the combined therapeutic strategy. This cell line was established from a primary adenocarcinoma of the colon and is characterized by an intrinsically high resistance to apoptosis that correlates with its resistance to 5-FU. 5-FU was absorbed into the matrix of the PCL NPs during synthesis using the interfacial polymer disposition method. The antitumor activity of gene E from the phage φX174 was tested by generating a stable clone (SW480/12/E). In addition, the localization of E protein and its activity in mitochondria were analyzed. We found that the incorporation of 5-FU into PCL NPs (which show no cytotoxicity alone), significantly improved the drug's anticancer activity, reducing the proliferation rate of colon cancer cells by up to 40-fold when compared with the nonincorporated drug alone. Furthermore, E gene expression sensitized colon cancer cells to the cytotoxic action of the 5-FU-based nanomedicine. Our findings demonstrate that despite the inherent resistance of SW480 to apoptosis, E gene activity is mediated by an apoptotic phenomenon that includes modulation of caspase-9 and caspase-3 expression and intense mitochondrial damage. Finally, a strongly synergistic antiproliferative effect was observed in colon cancer cells when E gene expression was combined with the activity of the 5-FU-loaded PCL NPs, thereby indicating the potential therapeutic value of the combined therapy.

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Enhancement of the Anti‐tumor Effect of 5′‐Deoxy‐5‐fluorouridine by Transfection of Thymidine Phosphorylase Gene into Human Colon Cancer Cells

Cited by 15 publications

References 21 publications

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

The role of thymidine phosphorylase, an angiogenic enzyme, in tumor progression

5-Fluorouracil-loaded poly(ε-caprolactone) nanoparticles combined with phage E gene therapy as a new strategy against colon cancer

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Enhancement of the Anti‐tumor Effect of 5′‐Deoxy‐5‐fluorouridine by Transfection of Thymidine Phosphorylase Gene into Human Colon Cancer Cells

Cited by 15 publications

References 21 publications

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase

The role of thymidine phosphorylase, an angiogenic enzyme, in tumor progression

5-Fluorouracil-loaded poly(&epsilon;-caprolactone) nanoparticles combined with phage E gene therapy as a new strategy against colon cancer

Contact Info

Product

Resources

About

5-Fluorouracil-loaded poly(ε-caprolactone) nanoparticles combined with phage E gene therapy as a new strategy against colon cancer