Serum vascular endothelial growth factor in epithelial ovarian neoplasms: correlation with patient survival.

OBJECTIVE: To investigate the relationship between serum vascular endothelial growth factor (VEGF) and clinicopathological factors and to determine whether VEGF is an independent prognostic factor of ovarian cancer patients. METHODS: Fifty-six women with International Federation of Gynecology and Obstetrics stages I to IV epithelial ovarian cancer undergoing surgery were enrolled. Clinical and pathologic items were recorded. Pretreatment VEGF serum samples of the 56 women were measured by an enzyme-linked immunosorbent assay. The results were correlated to clinical data. The histopathologic items and serum VEGF influencing clinical outcome were evaluated comparatively. RESULTS: The median VEGF serum level in ovarian cancer patients was 458.7 pg/mL. The 75% quatile was defined as the cutoff level. Elevated vascular endothelial growth factor serum levels before therapy correlated significantly with poorer disease-free survival (DFS) (log rank test, P = 0.001) and overall survival (OS) (log rank test, P < 0.001) on all of the 56 patients. Besides, significantly reduced DFS (log rank test, P = 0.001) and OR (log rank test, P = 0.006) were also observed on 40 patients with residual disease less than 2 cm. High histologic grade (RR = 2.24 for DFS; RR = 2.38 for OS) and elevated serum VEGF levels (RR = 3.34 for DFS; RR = 4.47 for OS) are the prognostic factors on the 56 ovarian carcinoma patients by multivariate analyses. The advanced surgical staging (RR = 3.28 for DFS; RR = 3.84 for OS), high histologic grade (RR = 2.55 for DFS; RR = 2.44 for OS), and elevated serum VEGF levels (RR = 5.62 for DFS; RR = 5.37 for OS) are the prognostic factors for 40 ovarian carcinoma patients with residual disease less than 2 cm by multivariate analyses. CONCLUSIONS: Pretreatment VEGF serum levels might be regarded as an additional factor in predicting the outcome of ovarian cancer patients. It also could provide prognostic information in clinically relevant subsets, such as those of residual disease less than 2 cm. Anti-angiogenic therapy, if is available, might be a new treatment modality for ovarian cancer patients with poor prognosis predicted by VEGF and other clinical parameters.     

Prognostic significance of vascular endothelial growth factor and its receptors in endometrial carcinoma

OBJECTIVE: The aim of this study has been to evaluate the clinical significance of expression of VEGF and its receptors, Flt-1, KDR, and Flt-4, in endometrial carcinomas. METHODS: Specimens of endometrial carcinomas from 86 patients were investigated immunohistochemically using respective specific antibodies. Additionally, samples from 14 patients with complex atypical endometrial hyperplasia (AEH) and 15 patients with normal endometria were also examined. Immunohistochemical assessment was classified as negative, weakly positive, and strongly positive according to staining intensity and the percentage of positive cells. RESULTS: In positive cases, VEGF and its receptors were usually expressed homogeneously in the cytoplasms of cells in the endometrial carcimona, similar to the staining intensity of endothelial cells of stromal microvessels adjacent to carcinoma nests. The overall positive rates in the 86 carcinoma specimens were 66% for VEGF, 51% for Flt-1, 38% for KDR, and 57% for Flt-4. Their expressions in endometrial carcinoma tissues were high with significance or with borderline significance, compared to those in samples of complex AEH or normal endometria. Survival curves determined by the Kaplan-Meier method and univariate analysis showed VEGF, Flt-1, and Flt-4 overexpression to be related to poor prognosis of patients with endometrial carcinomas. However, multivariate analysis revealed that Flt-4 overexpression correlated independently with poor survival, similar to a value for myometrial invasion at one-half or more and that for retroperitoneal lymph node metastasis, whereas VEGF and Flt-1 overexpression did not. CONCLUSION: Flt-4 overexpression might be a promising prognostic indicator for survival of a patient with endometrial carcinoma.     

卵巢上皮性癌组织中肿瘤浸润性树突状细胞的状态及其与血管内皮生长因子表达的相关性

目的研究卵巢上皮性癌组织中肿瘤浸润性树突状细胞(TIDC)的状态及其与血管内皮生长因子(VEGF)表达的关系。方法采用免疫组化链霉菌抗生物素蛋白-过氧化酶连接法和Picture二步法,检测57例卵巢上皮性癌(恶性组)、30例良性卵巢上皮性肿瘤(良性组)及16例正常卵巢(正常组)组织中S-100蛋白、CD83标记的TIDC状态及其VEGF的表达。结果(1)恶性组TIDC光镜下形态基本可分成两种,其分布有异质性。恶性组S-100^ TIDC中位数为4．3个／400高倍视野(HPF),分别与良性组(中位数为1．8个／HPF)和正常组(中位数为2．0个／HPF)比较,差异有显著性(P值分别为0．000和0．015)。恶性组中,早期(Ⅰ～Ⅱ期)肿瘤组织中S-100^ TIDC数量明显多于晚期(Ⅲ～Ⅳ期,中位数分别为6．0个和3．8个／HPF,P=0．026)。恶性组中,CD83^ TIDC浸润肿瘤间质者极少,中位数为0个／HPF。(2)恶性组细胞中VEGF高表达(5．0分),分别与良性组(3．8分)和正常组(2,4分)比较,差异均有极显著性(P=0.000)。(3)恶性组中,S-100^ TIDC数量与VEGF表达呈明显负相关(P=0．001)。结论(1)卵巢上皮性癌细胞可刺激TIDC的募集,但同时受VEGF的抑制。(2)卵巢上皮性癌中TIDC成熟严重受抑。     

Investigation of women with endometrial carcinoma using serum vascular endothelial growth factor (VEGF) measurement

Abstract. Gornall RJ, Anthony FW, Coombs EJ, Hogston P, Woolas RP. Investigation of women with endometrial carcinoma using serum endothelial growth factor (VEGF) measurement.
This study assessed whether serum VEGF measurement in women presenting with endometrial cancer could predict advanced stage disease. Preoperative sera from 37 women undergoing laparotomy for suspected endometrial cancer were assayed for VEGF, CA125 and platelet count. Significant positive correlation was shown between VEGF and platelet levels ( P = 0.003, r = 0.477). However, no correlation was demonstrated between VEGF and stage overall, and no significant difference was shown between those with early (stage 1A/1B, n = 20) compared to those with advanced (stage >1B, n = 13) or disseminated (stage >2, n = 7) disease. Serum VEGF measurement was not beneficial in the preoperative assessment of stage in patients with endometrial carcinoma. Strong correlation with platelet levels suggests that this is one of the sources of VEGF measured.     

Features associated with survival and disease-free survival in early endometrial cancer

Age, clinical stage, histologic grade, depth of myometrial penetration, adnexal status, peritoneal cytology, and progesterone and estrogen receptor status were available for 139 patients with clinical stage IA, IB, or II endometrial adenocarcinoma who had therapy at Indiana University Hospital or St. Vincent Hospital in Indianapolis. These features were analyzed for their association with survival and disease-free survival. Patients treated at Indiana University Hospital were similar to those from St. Vincent Hospital when comparisons were made by chi 2 test for age, clinical stage, grade, adnexal metastases, peritoneal cytologic results, progesterone receptor status, or estrogen receptor status. However, patients treated at Indiana University Hospital had lesions that were deeper (p = 0.03) than those treated at St. Vincent Hospital. Survival differences were observed for patients with progesterone receptor-rich versus progesterone receptor-poor tumors (p = 0.004), grades 1 and 2 versus grade 3 lesions (p = 0.013), and malignant versus benign peritoneal cytologic results (p = 0.01). Differences in disease-free survival were observed for those patients with adnexal metastases versus those with no adnexal disease (p = 0.002), those with estrogen receptor-rich versus estrogen receptor-poor tumors, outer third myometrial invasion (p = 0.002), and patients with clinical stage I versus clinical stage II disease (p = 0.03). A stepwise Cox proportional hazards model was constructed to determine correlates of disease-free survival. In the final model, grade (p = 0.0002), peritoneal cytologic results (p = 0.0002), progesterone receptor status (p = 0.004), and age as a continuous variable (p = 0.008) were most closely associated with disease-free survival.     

血清血管内皮细胞生长因子水平评价子宫颈癌预后的价值

目的探讨宫颈癌患者的血清血管内皮生长因子（VEGF）水平与宫颈癌临床病理特征及预后的关系。方法采用酶联免疫吸附法（ELISA）检测100例宫颈癌、50例宫颈上皮内瘤变患者和30例正常健康妇女血清中VEGF水平，并进行对照分析。结果宫颈癌患者的血清VEGF水平明显高于宫颈上皮内瘤变患者和健康妇女（P〈0.001），血清VEGF水平与患者的临床分期、分化程度、淋巴结转移、肿瘤大小及浸润深度显著相关（P〈0．001），而与病理类型无明显的相关（P〉0．05），术后血清VEGF平均含量较术前明显下降，术后复发血清VEGF含量较复发前明显升高，差异有显著性（P〈0．001）。结论检测血清VEGF对判断宫颈癌患者的肿瘤负荷、疗效、复发转移、预后都有重要的参考价值。     

Angiogenesis in endometrial carcinoma: correlation with survival and clinicopathologic risk factors

Association among angiogenesis, survival and clinicopathologic parameters in endometrial carcinoma was evaluated. Sixty patients who had been diagnosed as endometrial carcinoma, from 1993 to 1998, were included in the study. All patients had been surgically staged with bilateral pelvic and para-aortic lymph node dissection. All hysterectomy specimens were stained immunohistologically for factor VIII-related antigen. The area with the most intensified microvasculature was determined under low-power (x100) magnification, and the microvessel count of this area under high-power (x200) magnification was determined as the microvessel density (MVD) of the tumor. The mean MVD was 26.2 +/- 13.0 (range 6-68), and it was considered as high (n = 24; 40%), moderate (n = 19; 31.7%) and low (n = 17; 28.3%) when the MVD was >30, between 15-30 and <15, respectively. Statistical analysis included Mann-Whitney, Kruskal-Wallis and Spearman rank correlation tests. The Kaplan-Meier method was used to evaluate the difference between angiogenesis and survival. Multivariate analysis with the Cox regression model was used in MVD values and different clinicopathological parameters. There was positive correlation between MVD increase and surgicopathological stage (p < 0.05). A significant difference was seen between MVD increase and lymph node metastasis (p < 0.05). There were no differences between MVD and age, histological type, grade and lymphovascular invasion. MVD did not change in association with myometrial invasion depth. There was a significant difference in means of survival between the low and high MVD groups (p = 0.01). However, MVD was not an independent prognostic factor in multivariate analysis. Increased angiogenesis was found to be associated with advanced stage and decreased survival in endometrial carcinoma.     

Neutrophils infiltrating the endometrium express vascular endothelial growth factor: potential role in endometrial angiogenesis

OBJECTIVE(S): To identify leukocytes within the human endometrium expressing vascular endothelial growth factor (VEGF). DESIGN(S): Prospective cohort study. SETTING(S): Healthy volunteers in an academic research environment. PATIENTS(S): Twenty-one normal cycling women without abnormal menstrual bleeding or infertility. INTERVENTION(S): Endometrial tissue collection by Pipelle de Cornier aspiration. MAIN OUTCOME MEASURES(S): Histologic, immunohistochemical (CD3, CD34, CD56, CD68, neutrophil elastase, estrogen and P receptors, VEGF), and simultaneous double immunoenzymatic labeling analysis of VEGF-positive cells within the human endometrium. RESULT(S): Ten endometrial samples were obtained in the proliferative (cycle days 5-10) and 11 samples in the secretory phase (cycle days 15-26). Immunohistochemical analyses showed the expected distribution of the different leukocyte cell types. Besides epithelial and stromal endometrial cells, the predominant cells that stained for VEGF were neutrophil granulocytes. Neutrophils were more abundant in the secretory phase but they expressed neither estrogen-a nor P receptors. CONCLUSION(S): Neutrophil granulocytes infiltrating the human endometrium express VEGF and regulate cyclical endometrial vascular proliferation. Ovarian steroids indirectly influence neutrophil migration.     

Co-expression of vascular endothelial growth factor and thymidine phosphorylase in endometrial cancer.

OBJECTIVE: To examine expression of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) together with microvessel count in endometrial cancer, and to investigate the relationship with clinicopathological and biological factors. METHODS: VEGF expression, TP expression, the microvessel count (factor VIII-related antigen positive cells), bcl-2 expression, proliferating cell nuclear antigen (PCNA) index, and p53 expression were determined in 50 resected endometrial cancer specimens, using immunohistochemistry. The relationship between VEGF and TP expression and correlation between expression and microvessel count were also given attention. These 3 factors were analyzed with regard to clinicopathological factors, bcl-2 expression, PCNA index, and p53 expression. RESULTS: Staining status of VEGF and TP was identical in 37 (74%) of 50 tumors, the correlation being statistically significant (p < 0.01). Combined analysis of VEGF and TP status showed that tumors which were VEGF-positive and/or TP-positive had a significantly higher microvessel count than did tumors which were both VEGF-negative and TP-negative (p < 0.001 and p < 0.05, respectively). TP expression correlated with bcl-2 expression, and VEGF expression inversely correlated with the PCNA index. Although clinical stage (p < 0.01), PCNA index (p < 0.01), and p53 expression (p < 0.01) significantly correlated with disease-free survival, neither VEGF/TP expression nor microvessel count contributed to prognostic estimates. CONCLUSIONS: VEGF and TP may cooperatively promote angiogenesis in endometrial cancer, but these expressions may have limited additional prognostic value.     

Effect of vascular endothelial growth factor on sperm motility and survival

Vascular endothelial growth factor (VEGF) and its receptors are present in both male and female reproductive systems. In this experimental study, the effect of different concentrations of VEGF on sperm motility and survival in vitro was investigated. Human spermatozoa, collected from voluntary, proven fertile donors, were incubated in sperm washing medium containing different concentrations of VEGF (5, 10, 15, 20 ng/ml) for 24 h in a university reproductive endocrinology laboratory setting. Assessment of VEGF action on sperm motion characteristics was evaluated using a computer-assisted semen analyser. Sperm survival was determined by hypo-osmotic swelling and eosin-Y dye tests. VEGF had a positive effect on some parameters of sperm motility in a concentration-dependent manner. Maximal effect was observed at a concentration of 15 ng/ml; motility, progression, straight-line velocity and curvilinear velocity of VEGF-exposed spermatozoa were significantly increased (P < 0.05) at this concentration. However, sperm viability was not prolonged at any concentration of VEGF as shown by hypo-osmotic swelling and eosin-Y dye tests. VEGF may increase some sperm motility parameters, but not survival, in a concentration-dependent manner in vitro.     

血管内皮生长因子与血小板源性生长因子在卵巢上皮性癌淋巴管形成中的作用

目的 探讨血管内皮生长因子(VEGF)和血小板源性生长因子(PDGF)在卵巢上皮性癌(卵巢癌)淋巴管形成中的作用.方法 RT-PCR技术检测淋巴管内皮细胞核标志物Proxl和淋巴管形成相关因子VEGF-A、VEGF-C、VEGF-D及PDGF-A、PDGF-B、PDGF-C、PDGF-D在卵巢癌细胞株SKOV3、70例卵巢上皮性肿瘤(卵巢良性肿瘤15例、卵巢交界性肿瘤10例、卵巢癌45例)和20例正常卵巢组织中的表达情况.实时定量PCR技术检测卜述90例卵巢组织中Proxl、VEGF-A、-C、-D及PDGF-A、-B、-C、-D的表达水平,并进行相关性分析.结果 (1)Proxl在各种卵巢组织中均有表达,而在SKOV3细胞中无表达;VEGF-A、-C、-D及PDGF-A、-B、-C、-D在SKOV3细胞和各种卵巢组织中均有表达.(2)卵巢癌组织中Proxl(2.2±1.3)、VEGF.A(3.5±1.5)、VEGF-C(19 ±14)、VEGF-D(3.0±1.8)及PDGF-A(3.3±3.3)、PDGF-C(6.9±4.6)的表达水平高于卵巢良性肿瘤和交界性肿瘤(P均＜0.05).(3)Proxl、VEGF-A和PDGF-A在卵巢癌Ⅲ～Ⅳ期(Proxl:2.6±1.3,VEGF-A:4.0± 1.4.PDGF-A:4.1±3.7)、淋巴结转移阳性(Proxl:3.0±1.4,VEGF-A:4.1±1.7,PDGF-A:4.9±4.1)及腹膜转移阳性(Proxl:2.8±0.9,VEGF-A:4.0±1.8,PDGF-A:4.5±4.0)的组织中的表达水平,分别高于Ⅰ～Ⅱ期、淋巴结转移阴性和腹膜转移阴性者(P均＜0.05);VEGF-C、VEGF-D在淋巴结转移阳性卵巢癌组织中的表达水平(VEGF-c:24± 13,VEGF-D:3.9±2.0)高于淋巴结转移阴性者(P均＜0.05).(4)卵巢癌组织中Proxl的表达水平与VEGF-D(r=0.62,P＜0.01)、PDGF-C(r=0.91,P＜0.01)、PDGF-D(r=0.61,P＜0.01)的表达水平呈正相关关系.结论 VEGF-A、VEGF-C和PDGF-A可能通过参与淋巴管形成之外的机制促进卵巢癌的淋巴结转移;VEGF-D可以促进卵巢癌的淋巴管形成及淋巴结转移;PDGF-B与卵巢癌的淋巴管形成及淋巴结转移无关;PDGF-C、PDGF-D参与卵巢癌淋巴管形成,但无促进淋巴结转移的作用.     

Expression of vascular endothelial growth factor and assessment of microvascular density with CD 34 and endoglin in proliferative endometrium, endometrial hyperplasia, and endometrial carcinoma

The aim of this study was to compare vascular endothelial growth factor (VEGF), CD 34, and endoglin expressions as markers of angiogenesis in proliferative endometrium (PE), endometrial hyperplasia (EH), and endometrial carcinoma (EC) and to find the possible impact of angiogenesis on malign transformation. Formalin-fixed, paraffin-embedded tissues from 12 patients with PE, 23 patients with simple EH and complex EH with atypia, and 31 patients with EC were included. A semiquantitative scoring system was used to assess the intensity and degree of staining of VEGF. Microvessel density (MVD) was assessed with endoglin and anti-CD 34 in most vascular areas. VEGF expression was significantly higher in EC and EH than PE, but there was no difference between EC and EH. According to CD 34 staining, there were no differences in MVD between groups. However, mean MVD counts assessed by endoglin were significantly higher in EC than PE and EH. Although VEGF expression in EC was significantly higher, it did not correlate with other measures of angiogenesis. MVD using endoglin seemed to reflect neoplastic angiogenesis better than CD 34.     

Changes in vascular endothelial growth factor production associated with decidualization by human endometrial stromal cells in vitro

BACKGROUND: The aim of this study was to clarify changes in the expression of vascular endothelial growth factor (VEGF) during decidualization by endometrial stromal cells (ESCs) in vitro. METHODS: ESCs were separated by enzymic digestion and filtration, and were cultured with RPMI 1640 in 10% fetal calf serum (FCS) and treated with medroxyprogesterone acetate (MPA) and dibutyryl-cyclic adenosine monophosphate (db-cAMP) to induce decidualization in vitro. The levels of prolactin (PRL) in the culture media were measured by enzyme immunoassay (EIA) and the levels of VEGF were measured by enzyme-linked immunosorbent assay (ELISA). The expression of VEGF mRNA by ESCs and decidualized cells was analyzed by Northern blot analysis. RESULTS: In treated cells, PRL production was significantly increased due to treatment with both db-cAMP (0.5 mmol/L) and MPA (100 nmol/L). VEGF mRNA expression was detected without any stimulation by ESCs. VEGF production was also significantly greater in cells treated with db-cAMP (0.5 mmol/L) and MPA (1 nmol/L or 100 nmol/L) than in control cells. VEGF mRNA was also increased in association with decidualization in vitro. CONCLUSIONS: VEGF production increased in association with decidualization in this in vitro study. Decidualization of ESCs may play a role in the induction of growth in the human fetus and/or placentation. The mechanism involved may include influencing the production of angiogenic growth factors such as VEGF.     

Association study of vascular endothelial growth factor gene polymorphisms in endometrial carcinomas in a Japanese population

OBJECTIVE: Vascular endothelial growth factor (VEGF) is one of the most potent endothelial cell mitogens and plays a critical role in angiogenesis of endometrial carcinomas. Several studies have demonstrated positive associations between VEGF gene polymorphisms and several carcinomas. In this study we investigated whether VEGF gene polymorphisms are associated with endometrial carcinomas in a Japanese population. METHODS: The allele frequencies and genotype distributions of VEGF -460 C/T, +405 G/C, and +936 C/T polymorphisms were examined in 105 endometrial carcinomas and 179 controls using PCR-RFLP analysis. An association of these polymorphisms with three-year disease-free survival was evaluated using the Kaplan-Meier method. RESULTS: No significant differences in the allele frequencies and genotype distributions of VEGF -460 C/T (p = 0.54, 0.90), +405 G/C (p = 0.31, 0.17), and +936 C/T polymorphisms (p = 0.46, 0.24) were observed between endometrial carcinoma patients and controls. There were no significant differences in the frequencies of haplotype -460 T/+405 C between patients and controls. Futhermore, VEGF -460 C/T, +405 G/C, and +936 C/T polymorphisms were not associated with three-year disease-free survival of endometrial carcinoma patients. CONCLUSIONS: Although limited by sample size, our study did not demonstrated any evidence that VEGF -460 C/T, +405 G/C, and +936 C/T polymorphisms are associated with an increased risk of endometrial carcinomas in Japanese women.     

Elevated platelet count in patients with endometrial carcinoma: correlation with selected prognostic factors and with survival

Platelets may play a role in the metastatic process and, among gynecologic malignancies, thrombocytosis has been reported in cervical and ovarian malignancy. The present study was conducted in order to assess the prevalence of thrombocytosis in endometrial carcinoma and to correlate platelet count with prognostic factors and with survival. The prevalence of preoperative thrombocytosis was assessed in 66 endometrial carcinoma patients and their platelet count was correlated with selected prognostic factors and with projected survival. The prevalence of thrombocytosis ( 400 000 µL⁻¹) was low (one of 66 patients) compared with that in cervical and ovarian carcinoma. Nevertheless, a significant ( P = 0.032) correlation was found between an elevated ( 300 000 µL⁻¹) platelet count and unfavorable grade of differentiation. Patients with an elevated count also had a poorer survival rate and a higher prevalence of older age, high stage and deep myometrial invasion, but this trend did not reach statistical significance. The combination of unfavorable grade and an elevated platelet count had a higher specificity and positive predictive value for deep myometrial invasion than unfavorable grade alone. The prevalence of thrombocytosis in endometrial carcinoma is low. An elevated platelet count may have some prognostic significance, but its ultimate role in endometrial carcinoma remains to be elucidated.