The expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase associates with angiogenesis in epithelial ovarian cancer

Background The object of this study is to clarify the association of an angiogenic factor, PD-ECGF (platelet-derived endothelial cell growth factor/thymidine phosphorylase), with clinicopathologic factors, in this case tumor angiogenesis, in epithelial ovarian cancers. Methods Tumor specimens were obtained at the time of surgery from the primary lesion in 60 patients with epithelial ovarian cancer. Histologic cell types were assigned to tumors according to the World Health Organization classification: 26 were classified as serous adenocarcinoma, 15 as endometrioid adenocarcinoma, 9 as mucinous adenocarcinoma, 9 as clear cell carcinoma, and 1 as undifferentiated carcinoma. Surgical staging was based on the international Federation of Gynecology and Obstetrics (FIGO) staging system: 16 were stage I, 6 were stage II, 34 were stage III, and 4 were stage IV. Expression of PD-ECGF was evaluated by immunohistochemical staining. Microvessel density was assessed by immunostaining for factor VIII-related antigen in the most neovascularized area. Results Stroma cells stained more strongly than cancer cells (80% vs. 33%). The immunopositivity of PD-ECGF in stroma cells was higher in cases of advanced cancer. Expression of PD-ECGF in mucinous adenocarcinomas was significantly higher than that in serous adenocarcinomas, while PD-ECGF expression in clear cell carcinomas was significantly lower. The microvessel density in the cases with marked PD-ECGF-positive stroma cells was significantly higher than that in the cases with absent/minimal PD-ECGF-positive stroma cells (P<0.05). Conclusion The expression of PD-ECGF may play a crucial role in the promotion of angiogenesis in epithelial ovarian cancers.     

Angiogenic switch occurs during the precancerous stage of human esophageal squamous cell carcinoma

We previously reported that vascular endothelial growth factor (VEGF) expression correlates with vessel density in human esophageal squamous cell carcinomas. However, tumor angiogenesis is not controlled simply by the presence of VEGF, and is likely regulated by several angiogenic factors produced by tumor and host cells. The goal of the present study was to determine the angiogenic profile of precancerous and cancerous lesions of the esophagus. Expression of mRNAs for VEGF, platelet derived endothelial cell growth factor (PD-ECGF), basic fibroblast growth factor (bFGF), and interleukin (IL)-8 was examined in six esophageal carcinoma cell lines and fresh biopsy specimens from 16 patients with invasive esophageal carcinoma by RT-PCR. Immunohistochemical analyses with antibodies against VEGF, PD-ECGF, bFGF, and IL-8 were performed on archival specimens of 60 normal esophageal mucosa, 11 dysplasias and 49 carcinomas of the esophagus. Microvessels were stained with anti-CD34 antibody and quantified by counting the number of vessels in a x200 field in the most vascularized areas of the tumor. Esophageal carcinoma cell lines and tumor tissues expressed mRNAs for one or more these angiogenic factors at various levels. An initial increase in vessel density and enhanced expression of PD-ECGF and VEGF were observed in dysplastic epithelium. Vessel density was significantly higher in more advanced lesions. bFGF and IL-8 were not expressed in dysplasias and mucosal carcinomas, but expression was increased in late stage squamous cell carcinoma. These findings suggest that the angiogenic switch is a very early event in the development of invasive carcinoma. Several different angiogenic factors produced by tumor cells and host cells may regulate angiogenesis during different steps of esophageal carcinogenesis.     

Expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase in human gallbladder lesions

The aim of this study was to investigate the expression of platelet-derived endothelial growth factor (PD-ECGF) in human gallbladder carcinomas to elucidate its role in angiogenesis and tumour progression. To this end, 56 archival surgical specimens of gallbladder lesions were examined for PD-ECGF/thymidine phosphorylase (TP) expression by immunohistochemistry and the PD-ECGF/TP protein level was assessed in five fresh specimens of gallbladder carcinoma by enzyme-linked immunosorbent assay (ELISA). Hyperplastic epithelial cells and adenoma cells showed no or faint staining with PD-ECGF/TP. Out of 43 gallbladder carcinomas, 27 (63%) showed moderate to strong immunoreactivity in the cytoplasm and nuclei of the tumour cells. PD-ECGF/TP immunoreactivity in stromal infiltrating cells was detected in 43% (3/7) hyperplasias, 17% (1/6) adenomas and 86% (37/43) carcinomas. PD-ECGF/TP protein levels in carcinoma tissues were higher than those in corresponding normal mucosa. PD-ECGF/TP expression did not correlate with angiogenesis, but significantly correlated with depth of invasion, lymph node metastasis, and tumour stage. These results overall suggest that PD-ECGF/TP produced by both cancer cells and infiltrating cells is associated with tumour progression in human gallbladder carcinoma.     

Expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase in cervical intraepithelial neoplasia

Angiogenesis contributes to the growth and secondary spreading of solid tumors. Platelet-derived endothelial cell growth factor (PD-ECGF)/thymidine phosphorylase (TP) has been identified as such an angiogenic factor. In this study, the expression of PD-ECGF/TP and VEGF was evaluated by immunohistochemical staining of tumor specimens from 40 patients with cervical intraepithelial neoplasia (10 with moderate dysplasia; 10 with severe dysplasia; 10 with carcinoma in situ; 10 with invasive carcinoma). The microvessel density was assessed by immunostaining for factor VIII-related antigen in the most highly neovascularized area. In both the nucleus and cytoplasm, the intensity of PD-ECGF/TP expression in carcinoma in situ and invasive carcinoma was significantly stronger than that in moderate dysplasia. However, the intensity of VEGF expression was not significantly different in the various specimens. The microvessel density in mild dysplasia was significantly different from that in carcinoma in situ (p<0.05), and that in invasive carcinoma (p<0.05). There was no significant relationship between the microvessel density and the expression of PD-ECGF/TP or that of VEGF. These results show that the expression of PD-ECGF/TP appears to be involved in the promotion of angiogenesis in cervical intraepithelial neoplasia.     

Expression and histopathological correlation of CCR9 and CCL25 in ovarian cancer

Ovarian carcinoma is the most lethal gynecological malignancy among women and its poor prognosis is mainly due to metastasis. Chemokine receptor CCR9 is primarily expressed by a small subset of immune cells. The interactions between CCL25 and CCR9 have been implicated in leukocyte trafficking to the small bowel, a frequent metastatic site for ovarian cancer cells. We have previously shown that ovarian cancer cells express CCR9 and play an important role in cell migration, invasion and survival in the presence of its natural ligand in vitro. In this study, we have evaluated the expression of CCR9 and CCL25 in ovarian cancer cells and clinical samples. Ovarian cancer tissue microarrays from University of Alabama at Birmingham and AccuMax were stained for CCR9 and CCL25. Aperio ScanScope was used to acquire 80X digital images and expression analysis of CCR9 and CCL25. Flow cytometry and the Image stream system were used to conform the expression of CCR9 and CCL25 in ovarian cancer cells. Our results show significantly higher (p<0.001) expression of CCR9 and CCL25 in serous adenocarcinoma followed by serous papillary cystadenoma, endometrioid adenocarcinoma, mucinous adenocarcinoma, cystadenoma, mucinous boderline adenocarcinoma, clear cell carcinoma, granulosa cell tumor, dysgerminoma, transitional cell carcinoma, Brenner tumor, yolk sac tumor, adenocarcinoma and fibroma cases, compared to non-neoplastic ovarian tissue. Similar to tissue expression, CCR9 was also significantly expressed by the ovarian cancer cell lines (OVCAR-3 and SK-OV-3) in comparison to normal adult ovarian epithelial cell. We provide the first evidence that CCR9 and its natural ligand CCL25 are highly expressed by ovarian cancer tissue and their expression correlates with histological subtypes. Expression of this chemokine receptor and its ligand CCL25 within primary tumor tissue further suggests a potential role of this chemokine-receptor axis in ovarian cancer progression.     

Monocyte chemoattractant protein-1 expression correlates with macrophage infiltration and tumor vascularity in human gastric carcinomas

The purpose of this study was to investigate the role of interactions between tumor cells and macrophages during angiogenesis in human gastric carcinomas. Macrophage infiltration into tumors and monocyte chemoattractant protein-1 (MCP-1) expression was assessed in 72 archival specimens of gastric carcinoma for comparison with tumor vascularity. The mRNA expression of MCP-1 was examined by RT-PCR in 6 gastric carcinoma cell lines and in fresh biopsy specimens from 18 patients. Immunolocalization of representative angiogenic factors, vascular endothelial growth factor (VEGF), and platelet-derived endothelial cell growth factor (PD-ECGF) was also done. MCP-1 expression in tumor cells increased with the depth of tumor invasion (Tis 9.5%, T1 19.4%, T2-4 60.0%), as did microvessel density and macrophage infiltration. Macrophage counts correlated with vessel counts, and both were significantly higher in MCP-1-positive than in negative tumors. Of the 6 gastric carcinoma cell lines, 2 constitutively expressed MCP-1 mRNA. In 6 (33.3%) of 18 biopsy samples, MCP-1 mRNA was expressed at higher levels in tumor tissues than in normal mucosa. VEGF protein was expressed by gastric carcinoma cells, whereas PD-ECGF protein was expressed mainly by stromal mononuclear cells. MCP-1 expression correlated significantly with VEGF but not PD-ECGF expression in gastric carcinomas. These results suggest that MCP-1 produced by human gastric carcinoma cells plays a role in angiogenesis via macrophage recruitment and activation.     

Interleukin-10-mediated inhibition of angiogenesis and tumor growth in mice bearing VEGF-producing ovarian cancer

Interleukin-10 (IL-10) is an immunosuppressive cytokine produced by T lymphocytes and drawing attention as an inhibitor of tumor angiogenesis. In this study, we investigated antiangiogenic and tumor suppressive effects of IL-10 in ovarian cancer cells. mIL-10-expressing plasmid was transferred into two ovarian cancer cell lines, SHIN-3 [vascular endothelial growth factor (VEGF) producing] and KOC-2S (non-VEGF producing). After selection, mIL-10-expressing cells were obtained as SHIN-3/mIL-10 and KOC-2S/mIL-10. No significant differences were observed in in vitro growth properties between mIL-10-expressing cells and control (luciferase expressing) cells in either KOC-2S or SHIN-3. The angiogenic activities of mIL-10-expressing cells were measured by dorsal air sac assay, which detected the number of newly formed blood vessels within a chamber in vivo. In addition, tumor formation was evaluated by s.c. tumor transplantation, and survival was monitored after i.p. injection of ovarian cancer cells into BALB/c nude mice. Both in vivo angiogenic activity and tumor growth were significantly inhibited in SHIN-3/mIL-10 cells compared with the control. Moreover, peritoneal dissemination was inhibited, and the survival period was significantly prolonged (mean survival days > 90 versus 36). In contrast, in the case of KOC-2S cells, no significant differences were observed in any of the parameters tested. These results indicate that IL-10 has suppressive effects on angiogenesis, tumor growth, and peritoneal dissemination of VEGF-producing ovarian cancer cells. Although the mechanisms of the antiangiogenic effect of IL-10 are still unclear, the potential usefulness of IL-10-mediated gene therapy of ovarian cancer was suggested.     

Expression of thymidine phosphorylase correlates with microvessel density in prostate cancer

Angiogenesis in the growth and development of prostate cancer was the focus of this study. Various angiogenic factors and their clinicopathologic correlations with the progression of prostate cancer have been examined. Thymidine phosphorylase is identical to platelet-derived endothelial cell growth factor (TP/PD-ECGF) and has angiogenic activity. We investigated the expression of TP/PD-ECGF in prostate cancer and its association with angiogenesis or clinicopathologic findings in 81 cases with prostate cancer. Western blot analysis using a specific monoclonal antibody 654-1 revealed the existence of a 55 kDa TP/PD-ECGF protein in human prostate cancer tissue. Cancer tissue showed low-positive immunostaining in 32 cases (39.5%) and high positivity in 49 cases (60.5%). This protein expression indicated a statistically significant association with microvessel density (low vs. high TP/PD-ECGF expression group: mean +/- SD, 37.3+/-27.0 vs. 53.1+/-28.0 microvessels in three fields, p<0.05). No correlation was found between the expression of TP/PD-ECGF and nuclear grade, glandular differentiation, clinical stage or overall survival rate. TP/PD-ECGF may play an important role in tumor angiogenesis in prostate cancer tissues. Although the expression of TP/PD-ECGF was not correlated with clinical outcome in patients with prostate cancer, there remains the possibility that TP/PD-ECGF may support or modify the tumor growth through angiogenesis in cooperation with other factors.     

Angiogenesis and platelet-derived endothelial cell growth factor/thymidine phosphorylase expression in endometrial cancer.

The object of this study was to clarify the association of angiogenic factor, platelet-derived endothelial cell growth factor (PD-ECGF)/thymidine phosphorylase (dThdPase) with clinicopathological factors including tumor angiogenesis and patient outcome in endometrial cancer. There was no correlation between the expression of PD-ECGF in cancer cells and any of the clinicopathological variables. Immunopositivity for PD-ECGF in stroma cells was significantly higher in poorly differentiated adenocarcinomas. The microvessel counts correlated with PD-ECGF positive stroma cells (p<0.0001). Disease-free survival was significantly worse in patients with marked PD-ECGF expression in stromal cells and high microvessel count. A multivariate analysis using Cox's proportional hazard model showed that high microvessel counts independently predicted disease-free survival as well as stage and myometrial invasion. The expression of PD-ECGF in stroma cells may play a crucial role in the promotion of angiogenesis. Tumor angiogenesis can be used to predict prognosis in patients with endometrial cancer.     

TFF3 Expression as Stratification Marker in Borderline Epithelial Tumors of the Ovary

Borderline tumors (BOT) of the ovary account for 10% to 20% of ovarian neoplasms. Like ovarian cancer, BOT encompass several different histological subtypes (serous, mucinous, endometrioid, clear cell, transitional cell and mixed) with serous (SBOT) and mucinous (MBOT) the most common. Current hypotheses suggest low-grade serous carcinoma may develop in a stepwise fashion from SBOT whereas the majority of high grade serous carcinomas develop rapidly presumably from inclusion cysts or ovarian surface epithelium. The pathogenesis of mucinous ovarian tumors is still puzzling. Molecular markers could help to better define relationships between such entities. Trefoil factor-3 (TFF3) is an estrogen-regulated gene associated with prognosis in different types of cancer. It has also been included in a recent marker panel predicting subtypes of ovarian carcinoma. We analyzed the expression of TFF3 by immunohistochemistry in a cohort of 137 BOT and its association with histopathological features. Overall expression rate of TFF3 was 21.9%. None of the BOT with serous and endometrioid histology displayed strong TFF3 expression. On the other hand, TFF3 was highly expressed in 61.4% of MBOT cases and 33.3% of BOT with mixed histology (P < 0.001) suggesting a potential function of the protein in that subtypes. Associations of TFF3 expression with FIGO stage and micropapillary pattern were significant in the overall cohort but confounded by their correlation with histological subtypes. The highly specific expression of TFF3 in MBOT may help to further clarify potential relationships of tumors with mucinous histology and warrants further studies.     

DCC gene alterations in histological types and clinical stages of epithelial ovarian cancer

We examined loss of heterozygosity (LOH) and mRNA expression of the DCC gene in 77 tissues and 6 cell lines of human ovarian cancer. LOH was seen in serous and endometrioid adenocarcinomas but was not in clear cell and mucinous adenocarcinomas. LOH was exhibited in all clinical stages including stage I. In the DCC mRNA expression, 6 of 9 (66.7%) cancer tissues and all of 6 (100%) cancer cell lines showed loss or marked reduction. These results suggest that alterations of the DCC gene may play important roles in the pathway of carcinogenesis of human ovarian cancer, especially serous and endometrioid adenocarcinomas.     

WT1 and WT1-AS genes are inactivated by promoter methylation in ovarian clear cell adenocarcinoma

BACKGROUND: Ovarian clear cell adenocarcinoma is associated with one of the poorest prognoses among human epithelial ovarian cancers. The authors hypothesized that Wilms tumor suppressor 1 gene (WT1) sense and antisense (WT1-AS) expression and their promoter methylation status could characterize ovarian clear cell adenocarcinoma from ovarian serous adenocarcinoma. METHODS: To test this hypothesis, ovarian cancer cell lines and 42 cancer tissues (17 clear cell and 25 serous adenocarcinoma) were analyzed for expression and methylation of WT1 and WT1-AS genes. RESULTS: These experiments demonstrated that all serous adenocarcinoma tissues expressed both WT1 and WT1-AS genes, although expression of these genes was lacking in clear cell adenocarcinoma. The WT1 and WT1-AS promoter were significantly methylated in clear cell adenocarcinoma (88.2% and 88.2%, respectively) compared with serous adenocarcinoma (24.0% and 20.0%, respectively). Significant correlation between methylation and mRNA expression status was observed for each gene. Also in agreement with these data, WT1 and WT1-AS negative ovarian cancer cell lines reexpressed these genes after treatment with the demethylating agent, 5-aza-2'-deoxycytidine. CONCLUSIONS: The current study shows that CpG hypermethylation is an important mechanism of WT1 and WT1-AS gene inactivation in ovarian clear cell adenocarcinoma. This is the first report that has demonstrated differential expression and methylation of WT1-AS in ovarian clear cell and serous adenocarcinomas. This study presents new molecular characterizations between these two types of adenocarcinoma and may provide insight as to why clear cell adenocarcinoma has a poorer prognosis than serous adenocarcinoma of the ovary.     

53BP1基因在卵巢癌中的表达及其临床意义

目的:观察53BP1在不同类型卵巢癌(浆液性癌、透明细胞癌、子宫内膜样癌和黏液性癌)中的表达,探讨53BP1在卵巢癌中表达的临床意义.方法:应用免疫组化EnVision法研究157例卵巢癌(浆液性癌61例、透明细胞癌38例、子宫内膜样癌44例、黏液性癌14例)及65例正常卵巢组织中53BP1表达.结果:157例卵巢癌中,53BP1阳性表达者64例,阳性率为40.8％,而65例正常组织中,53BP1阳性率为70.8％,53BP1在卵巢癌组织中的阳性表达明显低于正常组织(P＜0.01).53BP1在不同类型卵巢癌中的阳性表达没有明显差异(P＞0.05).53BP1在卵巢Ⅱ型癌中的阳性表达高于Ⅰ型癌,但差异无统计学意义(P=0.058).53BP1在卵巢癌Ⅲ、Ⅳ期的阳性表达虽低于Ⅰ、Ⅱ期,但无统计学差异(P＞0.05).结论:53BP1失表达可能参与了卵巢癌的发生.     

Thymidine phosphorylase and vascular endothelial growth factor in patients with Stage I lung adenocarcinoma

BACKGROUND: Thymidine phosphorylase (TP) has chemotactic activity in endothelial cells in vitro and angiogenic activity in vivo. However, the clinical significance of TP and cooperative roles of TP with other angiogenic factors have remained unclear in nonsmall cell lung carcinoma (NSCLC). METHODS: The authors stained for TP, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) in 132 tumors from patients with Stage I NSCLC. They compared TP, VEGF, and bFGF expression levels with microvessel counts (MVCs), macrophage counts, mast cell counts, and clinical outcomes of patients with Stage I NSCLC. RESULTS: In adenocarcinoma samples, only stromal cell-TP expression and tumor cell-VEGF expression were associated with MVCs and mast cell counts but not with macrophage counts. In squamous cell carcinoma samples, there were no significant associations of the expression of any examined angiogenic factors with MVCs, mast cell counts, or macrophage counts. More importantly, only among patients with Stage I adenocarcinoma of the lung did patients in the stromal cell-TP positive tumor group and in the tumor cell-VEGF positive tumor group have a significantly worse prognosis compared with patients in the stromal cell-TP negative tumor group and in the tumor cell VEGF negative group, respectively. In addition, among patients with Stage I adenocarcinoma, patients in the stromal cell-TP positive and tumor cell-VEGF positive tumor group had a significantly worse prognosis among the four groups. CONCLUSIONS: TP induction in tumoral stroma, but not in tumor cells, and tumor cell-VEGF induction may promote angiogenesis cooperatively in adenocarcinoma of the lung. Stromal cell-TP expression and tumor cell-VEGF expression may be important prognostic factors in adenocarcinoma of the lung, and stromal cell-TP expression may be a marker of active remodeling stroma in adenocarcinoma of the lung.     

Expression of NEU3 (plasma membrane-associated sialidase) in clear cell adenocarcinoma of the ovary: its relationship with T factor of pTNM classification

Cell surface carbohydrate expression strongly influences the biological characteristics of cancer cells. Especially, it is known that the change of sialic acid expression could be related to the invasive and metastatic potentials of tumors. This study aimed to investigate sialidase expression of ovarian cancer cells and to evaluate the relationship between plasma membrane-associated sialidase (NEU3) expression and various clinicopathological factors in ovarian clear cell adenocarcinoma patients. In 18 cell lines derived from human ovarian cancers (including clear cell, mucinous, and serous adenocarcinoma), sialidase mRNA expression was evaluated by RT-PCR. NEU1 and NEU3 expression levels were found to be elevated in most cell lines while NEU2 and NEU4 expression was rarely elevated. Interestingly, NEU3 expression was detected in all clear cell adenocarcinoma cell lines. In 71 patients with ovarian clear cell adenocarcinoma, treated at Keio University Hospital from February 1983 to February 2002, NEU3 expression was examined by immunohistochemical staining of surgical specimens and clinicopathological factors were reviewed. NEU3 expression was found to be positive in 77.5% of all cases. Furthermore, a high level of NEU3 expression was significantly correlated with T3 factor of pTNM classification on univariate and multivariate analysis. This is the first report to show that NEU3 is expressed in most of ovarian clear cell adenocarcinoma. And our results show that NEU3 expression is correlated with T factor (pTNM classification) in ovarian clear cell adenocarcinoma.