Platelet-derived endothelial cell growth factor/thymidine phosphorylase expression correlated with tumor angiogenesis and macrophage infiltration in colorectal cancer

doi:10.1016/S0304-3835(98)00051-2

Cancer Letters

Volume 128, Issue 1, 5 June 1998, Pages 55-63

https://doi.org/10.1016/S0304-3835(98)00051-2 Get rights and content

Abstract

To clarify whether platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) expression in both tumor cells and stromal cells has independent or synergistic effects on tumor angiogenesis and progression and to explore a possible regulator for PD-ECGF/TP expression, immunohistochemical staining was conducted on 148 specimens of colorectal cancer. The microvessel count was significantly correlated with the extent of PD-ECGF/TP expression. Macrophage infiltration in tumors with positive TP was significantly higher than in tumors with negative TP (P<0.001). The Cox model showed that PD-ECGF/TP expression was an independent prognostic factor, although the microvessel count had a stronger value in determining the patient prognosis.

Introduction

The initiation and maintenance of tumor angiogenesis depends on the balance of angiogenic inducers and inhibitors [1]. A large effort has been made to identify pro- and anti-angiogenic molecules. Thymidine phosphorylase (TP), also known as platelet-derived endothelial cell growth factor (PD-ECGF) [2], is an enzyme that has been implicated indirectly in tumor angiogenesis 3, 4. TP activity has been reported to be increased considerably in tumor tissues relative to neighboring normal tissues in a variety of malignancies 5, 6, 7. PD-ECGF cDNA-transfected cells exhibited higher angiogenic activity than untransfected cells [3]. Experimental studies have revealed that the expression of PD-ECGF/TP in tumor cells is significantly correlated with intratumor microvessel density (IMVD) 8, 9, 10, 11, 12and hematogenous metastasis 10, 11. Moreover, several papers have reported an association between the expression of PD-ECGF/TP in tumor cells and the overall survival of patients with malignant tumors 9, 10. However, Takahashi et al. [13]showed that PD-ECGF/TP is predominantly expressed in tumor-infiltrating macrophages with only a minority of tumor cells (5%) staining positively in colorectal cancer. Fox et al. [14]reported that macrophages in most of the normal tissues were usually PD-ECGF/TP-positive with no apparent correlation with angiogenesis. As yet it is unknown whether the expression of PD-ECGF/TP in tumor stroma cells has any clinical or prognostic significance. The positive rates of PD-ECGF/TP expression in tumor cells and tumor stroma cells also remain to be investigated. In addition, a recent study showed that macrophage infiltration (MI) in breast carcinoma was correlated with tumor angiogenesis and prognosis [15]. Recent research revealed that PD-ECGF/TP in tumor cells can be upregulated by tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1) and interferon-γ (INF-γ) [16], suggesting that this enzyme is perhaps a mediator of angiogenesis in chronic inflammation and that tumor cells can amplify their own angiogenic activity by recruiting or activating macrophages. Accordingly, it is possible that PD-ECGF/TP expression may correlate with MI in tumor tissue.

To determine the positive rates of PD-ECGF/TP expression in either tumor cells or tumor stroma cells and to clarify whether tumor stromal PD-ECGF/TP expression has an independent or synergistic impact on tumor angiogenesis and patient prognosis, an immunohistochemical study using anti-PD-ECGF/TP antibody and anti-CD34, together with an assessment of macrophage infiltration, was conducted on resected specimens from 148 cases of colorectal cancer. The correlation of PD-ECGF/TP expression with tumor angiogenesis, MI and patient outcome was investigated.

Section snippets

Patients

A total of 148 patients with colorectal carcinomas who underwent surgery at our institution from 1983 to 1993 were studied. Patients had received neither chemotherapy nor radiotherapy before surgery. Pathological classification, including stage, invasion depth, histologic differentiation, lymphatic invasion and venous invasion, was defined in accordance with the General Rules for Clinical and Pathological Studies on Cancer of the Colon, Rectum and Anus by the Japanese Society for Cancer of the

Clinical outcome

Eighty-five (57%) of 148 patients survived with a median survival of 74 months (range 1–147 months), while the other 63 patients died within 2–109 months (median 30 months) after surgery. Hematogenous metastases were observed in 69 (47%) patients, which were diagnosed either at the time of surgery (17 cases) or during follow-up (52 cases).

Histological examination identified 69 (47%) cases with well-differentiated adenocarcinoma, 63 (42%) cases with moderately-differentiated adenocarcinoma,

Discussion

In this study, tumor cell PD-ECGF/TP expression was observed in 53% of patients with colorectal carcinoma. This finding contrasts with other reports using the same antibody. Takahashi et al. [13]reported that only 5% of colon cancer cases were stained positively. In a study on gastric cancer, Maeda et al. [10]reported that 60.8% of cases were positive for PD-ECGF/TP expression. This discrepancy may be explained by the difference in evaluation methods. Takahashi et al. [13]evaluated the

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Making capecitabine targeted therapy for breast cancer: Which is the role of thymidine phosphorylase?
2013, Clinical Breast Cancer
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Preclinical studies have shown that in nude mice, the tumor cells overexpressing TP grow faster and form more angiogenic tumors.32 In particular, upregulation of TP correlates with intratumoral microvessel density in human tumor tissue and with the expression of other proangiogenic factors such as vascular endothelial growth factor (VEGF) and HIF 2α.33–35 Thymidine phosphorylase and VEGF are two potent angiogenic molecules that seem to have a cooperative role in neovascularization and endothelial stimulation.
Thymidine phosphorylase (TP) expression has been found to be elevated in various solid tumors where it is likely involved in mechanisms that regulate cell proliferation, apoptosis, and angiogenesis. Based on these properties, it is tempting to hypothesize a potential prognostic role of TP, suggesting that a high TP expression could predict a poor outcome. On the other hand, TP expression has been studied for its role in predicting benefit from treatment with fluoropyrimidine-containing chemotherapy. Several studies have been conducted on breast cancer. The current evidence on the value of TP is not mature enough to allow its translation into clinical practice. However, several findings support the potentially predictive role of TP. In this light, TP appears to be a promising marker that can give helpful information to predict the benefit from capecitabine-based chemotherapy in patients with breast cancer.
Clinical, laboratory and molecular factors predicting chemotherapy efficacy and toxicity in colorectal cancer
2011, Critical Reviews in Oncology/Hematology
Colorectal cancer (CRC) treatment has evolved significantly over the last ten years with the use of active chemotherapeutic agents including fluoropyrimidines, oxaliplatin and irinotecan plus targeted monoclonal antibodies bevacizumab, cetuximab and panitumumab. The addition of newer chemotherapeutic agents and targeted therapies has improved patient outcomes at the cost of increased toxicity with not all patients benefiting from these treatments. It is necessary for clinicians to more accurately predict clinical outcomes particularly in the predominantly elderly CRC patient population. This review aims to summarise existing data regarding the use of clinical and laboratory variables plus molecular markers in predicting response, survival and toxicity to chemotherapy agents and targeted monoclonal antibodies currently used in the treatment of CRC.
The role of angiogenesis in solid tumours: An overview
2009, European Journal of Internal Medicine
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The most widely used angiogenic factor for this purpose is VEGF and a number of studies have demonstrated that tumour overexpression of VEGF correlates with high tumour MVD and is associated with advanced tumour stage or tumour invasiveness in various common human cancers [91,92]. Furthermore, the overexpression of other angiogenic factors such as PD-ECGF [93], bFGF [94], TGF-β [95], angiogenin [96], tissue factor [97] and COX-2 [98] in various cancers has been shown to correlate with advanced tumour stage and decreased patient survival. The circulating level of angiogenic factors has also been investigated as a surrogate marker of the angiogenic activity of tumours.
Angiogenesis is the physiological process of the formation of new blood vessels from pre-existing ones. Multiple molecules regulate angiogenesis, such as the vascular endothelial growth factor, angiopoietins, the fibroblast growth factor, the platelet-derived growth factor and the transforming growth factor-β. Angiogenesis plays an important role in the growth, progression and metastasis of a tumour. Inhibiting the angiogenic process or targeting existing tumour vessels can be used for treatment of tumours as an alternative or in parallel with conventional chemotherapy. Many anti-angiogenic factors are under investigation and some are already being used in clinical practice with various results.
A review on the use of molecular markers of cytotoxic therapy for colorectal cancer, what have we learned?
2009, European Journal of Cancer
Over the past decades, significant progress has been achieved in the cytotoxic treatment of colorectal cancer (CRC) by the use of fluoropyrimidines, irinotecan and oxaliplatin. However, as not all patients do respond to chemotherapy, there is a need for predictive and prognostic factors in order to optimise the treatment for individual patients. Although many potential molecular markers have been studied, none of these have been implemented in the standard of care for colorectal cancer patients.
We performed a review of the data on the prognostic and/or predictive value of molecular markers for cytotoxic drugs in CRC. The following markers were included: dihydropyrimidine dehydrogenase, orotate phosphoribosyl transferase, thymidine phosphorylase, thymidylate synthase, mismatch repair deficiency, topoisomerase 1, excision cross-complementing gene and carboxylesterases.
With the exception of mismatch repair deficiency, these molecular markers showed divergent and inconsistent results on their prognostic and/or predictive value. This underscores the complexity of the role of these markers.
We conclude that further retrospective testing of these markers is unlikely to add clinically useful results. More definite results may only be expected when these markers are included in the design of prospective randomised studies.
Radiation-Induced Thymidine Phosphorylase Upregulation in Rectal Cancer Is Mediated by Tumor-Associated Macrophages by Monocyte Chemoattractant Protein-1 From Cancer Cells
2009, International Journal of Radiation Oncology Biology Physics
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Most cells expressing TP in human colorectal cancer tissues have been reported to be stromal cells, especially TAMs (14). Alternatively, other studies have shown strong TP expression in cancer cells (15, 16). The observations that tumor cells stimulate the formation of stroma that secretes various mediators pivotal to the maintenance of the tumor microenvironment, and that TAMs are one of the major components of tumor stroma and are capable of promoting diverse aspects of tumor growth, favor the expression of TP in the stromal cells, however (17).
The mechanisms of thymidine phosphorylase (TP) regulation induced by radiation therapy (XRT) in various tumors are poorly understood. We investigated the effect and mechanisms of preoperative XRT on TP expression in rectal cancer tissues.
TP expression and CD68 and monocyte chemoattractant protein–1 (MCP-1) levels in rectal cancer tissues and cancer cell lines were evaluated before and after XRT in Western blotting, immunohistochemistry, enzyme-linked immunoassay, and reverse transcription–polymerase chain reaction studies. Isolated peripheral blood monocytes were used in the study of chemotaxis under the influence of MCP-1 released by irradiated colon cancer cells.
Expression of TP was significantly elevated by 9 Gy of XRT in most rectal cancer tissues but not by higher doses of XRT. In keeping with the close correlation of the increase in both TP expression and the number of tumor-associated macrophages (TAMs), anti-TP immunoreactivity was found in the CD68-positive TAMs and not the neoplastic cells. Expression of MCP-1 was increased in most cases after XRT, and this increase was strongly correlated with TP expression. However, this increase in MCP-1 expression occurred in tumor cells and not stromal cells. The XRT upregulated MCP-1 mRNA and also triggered the release of MCP-1 protein from cultured colon cancer cells. The supernatant of irradiated colon cancer cells showed strong chemotactic activity for monocyte migration, but this activity was completely abolished by neutralizing antibody.
Use of XRT induces MCP-1 expression in cancer cells, which causes circulating monocytes to be recruited into TAMs, which then upregulate TP expression in rectal cancer tissues.
Activity and substrate specificity of pyrimidine phosphorylases and their role in fluoropyrimidine sensitivity in colon cancer cell lines
2007, International Journal of Biochemistry and Cell Biology
Thymidine phosphorylase (TP) and uridine phosphorylase (UP) are often upregulated in solid tumors and catalyze the phosphorolysis of natural (deoxy)nucleosides and a wide variety of fluorinated pyrimidine nucleosides. Because the relative contribution of each of the two enzymes to these reactions is still largely unknown, we investigated the substrate specificity of TP and UP in colon cancer cells for the (fluoro)pyrimidine nucleosides thymidine (TdR), uridine (Urd), 5′-deoxy-5-fluorouridine (5′DFUR), and 5FU. Specific inhibitors of TP (TPI) and UP (BAU) were used to determine the contribution of each enzyme in relation to their cytotoxic effect.
The high TP expressing Colo320TP1 cells were most sensitive to 5′DFUR and 5FU, with IC₅₀ values of 1.4 and 0.2 μM, respectively, while SW948 and SW1398 were insensitive to 5′DFUR (IC₅₀ > 150 μM for 5′DFUR). TPI and BAU only moderately affected sensitivity of Colo320, SW948, and SW1398, whereas TPI significantly increased IC₅₀ for 5′DFUR (50-fold) and 5FU (11-fold) in Colo320TP1 and BAU that in C26A (9-fold for 5′DFUR; p < 0.01). In the epithelial skin cell line HaCaT both inhibitors were able to decrease sensitivity to 5′DFUR and 5FU separately. HaCaT might be a model for 5′DFUR toxicity.
In the colon cancer cells 5′DFUR degradation varied from 0.4 to 50 nmol 5FU/h/10⁶ cells, that of TdR from 0.3 to 103 nmol thymine/h/10⁶ cells, that of Urd from 0.8 to 79 nmol uracil/h/10⁶ cells, while conversion of 5FU to FUrd was from 0.3 to 46 nmol/h/10⁶ cells. SW948 and SW1398 were about equally sensitive to 5′DFUR and 5FU, but SW1398 had higher phosphorylase activity (>65-fold) compared to SW948.
In SW948 and HaCaT TPI and BAU inhibited TdR and Urd phosphorolysis (>80%), respectively. Both TP and UP contributed to the phosphorolysis of 5′DFUR and 5FU. In the presence of both inhibitors, still phosphorolysis of 5FU (>40%) was detected in the tumor and HaCaT cell lines, and remarkably, that of all four substrates in SW1398 cells.
5′DFUR phosphorolysis was also measured in situ, where Colo320TP1, SW1398, and HaCaT cells produced significant amounts 5FU from 5′DFUR (>10 nmol/24 h/10⁶ cells). In Colo320TP1 and in HaCaT cells TPI completely prevented 5FU production, but not in SW1398 cells, where BAU decreased this by 67% (p < 0.01). High uracil and dUrd levels were detected in the medium. Uracil accumulation was heavily reduced in the presence of TPI for Colo320TP1 and HaCaT cells, whereas 5FU-induced dUrd production by these cell lines increased (p < 0.01). In contrast, for SW1398 cells only BAU was able to reduce uracil levels, and dUrd production remained unchanged.
In conclusion, overlapping substrate specificity was found for TP and UP in the cell lines, in which both enzymes were responsible for converting TdR and Urd, and 5′DFUR. 5′DFUR and 5FU were converted to their products in both the colon cancer cells and keratinocytes.

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Platelet-derived endothelial cell growth factor/thymidine phosphorylase expression correlated with tumor angiogenesis and macrophage infiltration in colorectal cancer

Abstract

Introduction

Section snippets

Patients

Clinical outcome

Discussion

Nature

J. Biol. Chem.

Biochim. Biophys. Acta

Cancer Lett.

Angiogenic factor (letter)

Nature

Identification of angiogenic activity and the cloning and expression of platelet-derived endothelial cell growth factor

Nature

Angiogenic activity of enzymes (letter)

Nature

Elevated thymidine phosphorylase activity in the plasma and ascitic fluids of tumor

Proc. Soc. Exp. Biol. Med.

Expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase in human breast cancer

Int. J. Cancer

Clinicopathologic and prognostic significance of an angiogenic factor, thymidine phosphorylase, in human colorectal carcinoma

J. Natl. Cancer Inst.

Thymidine phosphorylase/platelet-derived endothelial cell growth factor expression associated with hepatic metastasis in gastric carcinoma

Br. J. Cancer

Platelet-derived endothelial cell growth factor expression correlates with tumour angiogenesis and prognosis in non-small-cell lung cancer

Br. J. Cancer