Increased sensitivity to the prodrug 5'-deoxy-5-fluorouridine and modulation of 5-fluoro-2'-deoxyuridine sensitivity in MCF-7 cells transfected with thymidine phosphorylase

Patterson, Adam V.; Zhang, Haodong; Moghaddam, Amir; Bicknell, Roy; Talbot, D C; Stratford, I J; Harris, Adrian L.

doi:10.1038/bjc.1995.392

Cited by 115 publications

(90 citation statements)

References 8 publications

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“…Using strong virus promoters, it has been reported that the 5-FU derived from 5 0 -DFUR can diffuse from the transfected cancer cells to exert bystander killing effect on their neighbour cells in a gap junction-independent manner (Patterson et al, 1995;Kato et al, 1997). In this study, we also compared the bystander effect of pFBD/KCT-and pcDNA3/TP-transfected cancer cell lines.…”

Section: Discussionmentioning

confidence: 96%

“…Low transfection rate is still one of the major barriers for the success of cancer GDEPT in the clinic. It has been reported that the small portion of CMV promoter-driven TP-transfected cells can effectively convert 5 0 -DFUR into 5-FU, which can be released and performed cytotoxic effect onto the neighbouring cells (bystander effect) (Patterson et al, 1995). Figure 5C and D show that when pcDNA3-TP-or pFBD/KCT-transfected RKO or H630 cells composed only 5% of the whole-cell population, the sensitivity of H630 and RKO cell lines to 5 0 -DFUR was significantly enhanced.…”

Section: The Bystander Effect Of Pfastbacdualdpromoter/jb4-cea205-tp-mentioning

confidence: 99%

“…Consequently, insufficient conversion of 5 0 -DFUR into 5-FU in tumour tissues becomes the major limitation of CAP-based chemotherapy. Strong promoters such as CMV have been used to increase CD and TP expression in cancer cells and demonstrate enhanced cytotoxicity of 5 0 -DFCR and 5 0 -DFUR in vitro and in vivo (Huber et al, 1994;Patterson et al, 1995;Evrard et al, 1999;Ciccolini et al, 2001;Manome et al, 2001). Yet, the lack of specificity of these promoters limits the clinical development of GDEPT strategies.…”

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confidence: 99%

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In vitro evaluation of cancer-specific NF-κB-CEA enhancer–promoter system for 5-fluorouracil prodrug gene therapy in colon cancer cell lines

et al. 2007

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Nuclear factor-kappa B (NF-kB) is a transcription factor with high transcriptional activity in cancer cells. In this study, we developed a novel enhancer -promoter system, kB4-CEA205, in which the basal carcinoembryonic antigen (CEA) promoter sequence (CEA205) was placed downstream of the four tandem-linked NF-kB DNA-binding sites (kB4). In combination with a kB4 enhancer, the transcriptional activity of the CEA promoter was significantly enhanced (three-to eight-fold) in cancer cell lines but not in normal cells. In cancer cell lines, the transcriptional activity of kB4-CEA205 was comparable with that of the SV40 promoter. We also constructed vectors in which the thymidine phosphorylase (TP) cDNA was under the control of CEA205, kB4, kB4-CEA205 and CMV promoters, respectively. TP protein and enzyme activity were detected at comparable levels in kB4-CEA205-and CMV-driven TP cDNAtransfected cancer cell lines (H630 and RKO). The kB4-TP and CEA205-TP-transfected cell lines, respectively, only demonstrated negligible and low levels of TP protein and enzyme activity. Both CMV-and kB4-CEA205-driven TP cDNA transiently transfected cells were 8-to 10-fold sensitised to 5-fluorouracil (5-FU) prodrug, 5 0 -deoxy-5-fluorouradine (5 0 -DFUR), in contrast to only 1.5-to 2-fold sensitised by the kB4-and CEA205-driven TP cDNA-transfected cells. The bystander killing effect of CMV-and kB4-CEA205-driven TP cDNA-transfected cells was comparable. This is the first report that indicates that the NF-kB DNA-binding site could be used as a novel cancer-specific enhancer to improve cancer-specific promoter activity in gene-directed enzyme prodrug therapy. Lack of selectivity for malignant cells leading to dose-limiting toxicity is one of the major obstacles for the success of cancer chemotherapy. Gene-directed enzyme prodrug therapy (GDEPT) is potentially an elegant way to improve the therapeutic index of active anticancer drugs, but one of the major limitations for the clinical application of this approach is the lack of strong cancerspecific promoters (Aghi et al, 2000;Rooseboom et al, 2004).Carcinoembryonic antigen (CEA) is not expressed in normal and overexpressed in epithelial cancer cells (Shively and Beatty, 1985). Four cis-acting DNA-binding elements mapped on the CEA promoter region within 300 bp upstream of CEA translation starting point, especially, the first three elements are essential for specific CEA transcription (Hauck and Stanners, 1995;Richards et al, 1995a). Utilisation of the CEA promoter in an adenovirus vector for cytosine deaminase (CD) gene expression improves the selectivity of 5 0 -deoxy-5-fluorocytidine (5-DFCR)/5-fluorouracil (5-FU) conversion in CEA-expressing tumours (Richards et al, 1995a;Lan et al, 1996a). However, the transcriptional activity of CEA in most cancer cell lines is 10-to 300-fold lower than that of CMV and RSV viral promoters (Lan et al, 1996b;Ueda et al, 2001).Even when using adenovirus to deliver CEA promoter-driven suicide gene in vivo, few reports have demonstrated sufficient antitumou...

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Section: Discussionmentioning

confidence: 96%

Section: The Bystander Effect Of Pfastbacdualdpromoter/jb4-cea205-tp-mentioning

confidence: 99%

mentioning

confidence: 99%

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In vitro evaluation of cancer-specific NF-κB-CEA enhancer–promoter system for 5-fluorouracil prodrug gene therapy in colon cancer cell lines

et al. 2007

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“…Not only did TP promote endothelial cell migration, but cancer cells overexpressing the factor had a significantly faster growth rate . TP enzyme activity is also essential for the metabolism of fluoropyrimidines, which suggests that TP-overexpressing tumours may be sensitive to antimetabolite chemotherapy (Patterson et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

Different patterns of stromal and cancer cell thymidine phosphorylase reactivity in non-small-cell lung cancer: impact on tumour neoangiogenesis and survival

Koukourakis

Giatromanolaki²,

Kakolyris³

et al. 1998

Br J Cancer

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113

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Summary Angiogenesis is recognized as an important step in tumour pathogenesis that is related to invasion and metastatic spread and which consequently results in poor clinical outcome. In this study, we have examined the role of tumour stroma-activated fibroblasts and macrophage infiltration in the development of the angiogenic and metastatic phenotype in non-small-cell lung cancer (NSCLC). A total of 141 cases of early stage I-Il NSCLC treated with surgery alone were analysed. The JC-70 (anti-CD31) MAb was used for the assessment of vascular grade. The P-GF.44C MAb was used to assess thymidine phosphorylase (TP) reactivity in cancer cells, stromal fibroblasts and macrophages. Cancer cell TP overexpression related to high vascular grade and to advanced T stage (P = 0.0004 and P = 0.02). Expression of TP in stromal fibroblasts also correlated with high angiogenesis (P = 0.01), but was independent of cancer cell expression. Fibroblast TP overexpression was related to abundant stroma (P = 0.003), suggesting that TP may be a marker of active stroma. Moreover, intense macrophage infiltration was associated with fibroblast TP reactivity, regardless of the amount of stroma, suggesting that macrophages may be a major contributor to TP expression in stroma. Survival analysis showed that cancer cell TP overexpression was related to poor prognosis (P = 0.005). Although stroma TP is related to angiogenesis, in the low vascular grade group it defined a group of patients with better prognosis (P = 0.02). It may be that fibroblast TP reactivity is an indirect marker of tumour infiltration by functional macrophages, which have an antitumour effect. We conclude that stromal macrophage and fibroblast TP reactivity may have an important role in non-small-cell lung cancer behaviour. Understanding the role of stromal fibroblasts and inflammatory cells and their interaction with oncoprotein expression is essential for the elucidation of lung cancer pathogenesis.

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“…TP has been shown to exhibit a chemotactic and mitogenic capacity on endothelial cells in several angiogenic model systems, and its expression in human breast cancer cells has been shown to correlate with microvessel density in some studies (Fajardo et al, 1992;Folkman, 1996;Fox et al, 1996). Moreover, TP expression is 260-fold higher in invasive bladder cancer (O'Brien et al, 1995) and 27-fold higher in invasive breast carcinoma than normal tissue (Patterson et al, 1995). In ovarian carcinomas, areas of increased expression of TP have been associated with high blood velocity as measured by colour Doppler imaging (Reynolds et al, 1994).…”

mentioning

confidence: 99%

Association of tumour necrosis factor alpha and its receptors with thymidine phosphorylase expression in invasive breast carcinoma

Leek

Landers²,

Fox³

et al. 1998

Br J Cancer

Self Cite

192

118

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Summary Angiogenesis is an essential requirement for tumour growth and metastasis and is regulated by a complex network of factors produced by both stromal cells and neoplastic cells within solid tumours. The cytokine tumour necrosis factor alpha (TNF-ca) and the enzyme thymidine phosphorylase (TP) are two factors known to promote tumour angiogenesis. We have demonstrated recently that high numbers of tumour-associated macrophages (TAMs) are significantly associated with increased tumour angiogenesis and poor prognosis in invasive carcinoma of the breast. We have also shown that TAMs are a major source of TNF-a in invasive breast carcinomas, and that macrophagelike stromal cells as well as tumour cells synthesize TP in such tumours. However, little is known of the factors that regulate the production or activity of these factors in the tumour microenvironment. As TNF-a has been shown to up-regulate TP expression in tumour cells in vitro we performed an immunohistochemical study to investigate the possibility that TNF-a may be involved in the regulation of TP expression by malignant breast epithelial cells in vivo. To do this, we used a cocktail of non-neutralizing monoclonal anti-TNF-a antibodies to visualize both TNF-a-expressing macrophages and TNF-a bound to its receptors on tumour cells and endothelial cells in a series of 93 invasive carcinomas of the breast. A semiquantitative grading system was then used to compare these staining patterns with that for TP in the same biopsies. TNF-a immunoreactivity was also compared with various important tumour variables known to relate to outcome in this disease (microvessel density, node status, grade, stage, receptor status and macrophage infiltration), as well as relapse-free and overall survival data for these patients. Our data show significant positive correlations between TNF-a bound to its receptors on tumour cells and: (1) TP protein production by tumour cells, and (2) axillary lymph node status (i.e. metastasis). These results suggest that tumour cell responsiveness to TNF-a produced by neighbouring TAMs may play a part in the regulation of TP expression by tumour cells as well as their metastatic behaviour. This may explain, in part, the relationship between increased macrophage infiltration and angiogenesis in breast cancer, and further supports the contention that TAMs may represent an important target for future anti-angiogenic therapies.

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Increased sensitivity to the prodrug 5'-deoxy-5-fluorouridine and modulation of 5-fluoro-2'-deoxyuridine sensitivity in MCF-7 cells transfected with thymidine phosphorylase

Cited by 115 publications

References 8 publications

In vitro evaluation of cancer-specific NF-κB-CEA enhancer–promoter system for 5-fluorouracil prodrug gene therapy in colon cancer cell lines

In vitro evaluation of cancer-specific NF-κB-CEA enhancer–promoter system for 5-fluorouracil prodrug gene therapy in colon cancer cell lines

Different patterns of stromal and cancer cell thymidine phosphorylase reactivity in non-small-cell lung cancer: impact on tumour neoangiogenesis and survival

Association of tumour necrosis factor alpha and its receptors with thymidine phosphorylase expression in invasive breast carcinoma

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