Variations in serum vascular endothelial growth factor binding profiles and the development of ovarian hyperstimulation syndrome

OBJECTIVE: To compare the ability of serum to sequester vascular endothelial growth factor (VEGF) among patients who did and did not develop ovarian hyperstimulation syndrome (OHSS). DESIGN: Prospective, observational study. SETTING: A regional fertility centre with a commitment to research. PATIENT(S): Five patients undergoing controlled ovarian hyperstimulation as part of an in vitro fertilization cycle who developed severe OHSS, and five controls. INTERVENTION(S): Serum, collected at the time of oocyte retrieval, was incubated with radioactive VEGF (125I-VEGF(165)) for 2 hours before being passed down a sephadex G-150 gel filtration column. The fractional radioactive profile was then determined. MAIN OUTCOME MEASURE(S): The distribution of radioactive VEGF across the various fractions was measured in serum samples obtained from the two groups. RESULT(S): The 125I-VEGF(165) applied to the column eluted in two peaks centered on fractions 18 +/- 2 and 39 +/- 2. The molecular weight in the first peak was >300,000 kDa and represented "bound" VEGF; the second peak represented "unbound" VEGF. In the OHSS group, there was statistically significantly less radioactivity in the first peak than in the no-OHSS group. CONCLUSION(S): Patients who do not develop OHSS appear to have a high-molecular-weight protein that binds VEGF to a greater degree than occurs in patients who develop OHSS.     

Serum vascular endothelial growth factor levels are poorly predictive of subsequent ovarian hyperstimulation syndrome in highly responsive women undergoing assisted conception

OBJECTIVE: To determine whether serum vascular endothelial growth factor (VEGF) levels can distinguish highly responsive women who subsequently develop ovarian hyperstimulation syndrome (OHSS) from women with a similar ovarian response who do not. DESIGN: Prospective controlled study. SETTING: University IVF unit. PATIENT(S): Women undergoing IVF who met predetermined risk criteria for OHSS. Patients developing OHSS were compared with patients who did not develop OHSS. INTERVENTION(S): Long-protocol pituitary down-regulation followed by FSH stimulation by a standard protocol without coasting. A maximum of three embryos was transferred. Vaginal progesterone was used for luteal support. MAIN OUTCOME MEASURE: Occurrence of OHSS; serum VEGF concentrations on the day of embryo transfer (ET) and at 5 and 10 days after ET. RESULTS: Serum VEGF levels at any time point did not differ significantly between 9 OHSS cases and 9 controls. Vascular endothelial growth factor levels in samples collected from cases before the onset of OHSS were higher than levels in time-matched samples from controls (medians, 177.6 [range, 64.02-549.1] pg/mL vs. 100.7 [range, 37.59-246] pg/mL, respectively), but the difference failed to reach statistical significance (P=.0518), and there was considerable overlap between cases and controls. CONCLUSIONS: Serum VEGF levels in the luteal phase do not distinguish between high-risk women who subsequently develop OHSS and women with a similar risk profile who do not develop OHSS.     

Effect of GnRH agonist and hCG treatment on VEGF, angiopoietin-2, and VE-cadherin: trying to explain the link to ovarian hyperstimulation syndrome

This study evaluated the differentially modulated expression of vascular mediators in oocyte donors after hCG vs. GnRHa triggering, trying to understand ovarian hyperstimulation syndrome pathophysiology. Donors who received GnRH agonist triggering showed a statistically significant decrease in vascular endothelial growth factor in follicular fluid and in mRNA expression in granulosa cells, with no differences in angiopoietin 2 and vascular endothelial cadherin levels in serum or follicular fluid. This differential regulation of vascular endothelial growth factor by hCG might explain the higher likelihood of ovarian hyperstimulation syndrome following hCG administration compared with GnRH agonists.     

Increased risk of ovarian hyperstimulation syndrome following controlled ovarian hyperstimulation in patients with vascular endothelial growth factor +405 cc genotype

Ovarian hyperstimulation syndrome (OHSS) is a serious complication following controlled ovarian hyperstimulation (COH) for in vitro fertilization. OHSS has a range of clinical features from mild abdominal distention to severe thromboembolic events. Several clinical manifestations of OHSS such as ascites and hemoconcentration can be attributed to increased vascular permeability. Vascular endothelial growth factor (VEGF) and its receptor VEGFR2 have been identified as an important signaling system in mediating this increase. There is considerable genetic variation in the VEGF/R2 signaling system. We present the first study to examine if single nucleotide polymorphisms (SNPs) in the genes encoding the VEGF/R2 signaling system are associated with OHSS following COH. Blood samples from 53 OHSS patients and 100 controls were analyzed for six SNPs of interest. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated by a multivariate logistic regression model. We found an association between the VEGF +405cc genotype and OHSS (OR 3.4, 95% CI 1.01–11.7). This finding requires confirmation from other patient populations.     

Influence of severe endometriosis on gene expression of vascular endothelial growth factor and interleukin-6 in granulosa cells from patients undergoing controlled ovarian hyperstimulation for in vitro fertilization-embryo transfer

OBJECTIVE: To evaluate how endometriosis affects expression of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) in granulosa cells. DESIGN: Prospective study. SETTING: IVF-ET program at Osaka Medical College. PATIENT(S): Seventeen patients with revised American Fertility Society stage IV endometriosis and 17 patients with tubal infertility and no endometriosis. INTERVENTION(S): Granulosa cells obtained at oocyte retrieval were examined for VEGF and IL-6 gene expression. MAIN OUTCOME MEASURE(S): Serum E(2) and P levels at hCG administration, number of oocytes, fertilization rate, high-quality embryo rate, and pregnancy rate, and expression of VEGF and IL-6 genes. RESULT(S): Total hMG and FSH levels were statistically significantly higher in patients with endometriosis; however, the number of retrieved oocytes and the fertilization rate were lower compared with patients with tubal infertility. Serum E(2) levels and expression of VEGF in patients with tubal infertility were statistically significantly higher than those in patients with endometriosis. Interleukin-6 gene expression did not differ between the groups. CONCLUSION(S): In severe endometriosis, lower VEGF gene expression in granulosa cells may adversely affect oocyte development and maturation.     

血管内皮生长因子在预测卵巢过度刺激综合征中的作用

目的探讨在体外受精-胚胎移植(IVF-ET)周期中血清及卵泡液中血管内皮生长因子(VEGF)水平的变化,及其在预测卵巢过度刺激综合征(OHSS)中的作用.方法收集42例行IVF-ET患者的血清和卵泡液标本,采用定量酶联免疫方法检测血清、卵泡液中VEGF水平,按是否发生OHSS分为OHSS组(10例)及对照组(32例),对两组临床资料、性激素和VEGF水平进行回顾性相关分析.结果 OHSS组血清VEGF水平在月经周期的各个时期均高于对照组,但差异无显著性(P>0.05).OHSS组卵泡液VEGF水平为(1 487.7±365.8) ng/L,较对照组的(1 025.8±474.7) ng/L明显升高;取卵时卵泡数目增多,OHSS组为(10.0±5.9)个,对照组为(6.1±2.3)个;基础黄体生成素(LH),人绒毛膜促性腺激素(hCG)、注射日雌二醇(E2)水平均比对照组高,差异有显著性(P<0.05).结论 OHSS组卵泡液VEGF水平明显高于对照组,提示VEGF可能参与了OHSS的发病;卵泡液VEGF水平可以作为预测OHSS发生的检测指标.     

血管内皮生长因子在卵巢过度刺激综合征发病机理中作用的初步研究

目的通过对卵巢过度刺激综合征(OHSS)患者血管内皮生长因子(VEGF)水平和表达的测定,初步探讨VEGF在OHSS发病机理中的作用.方法采用酶联免疫吸附试验方法对14例中、重度OHSS患者(OHSS组)和同期13例非OHSS患者(对照组)取卵日卵泡液、人绒毛膜促性腺激素(hCG)注射日、胚胎移植(ET)日血清VEGF水平进行测定;采用免疫组织化学方法和逆转录聚合酶链反应技术检测取卵日卵巢黄素化颗粒细胞VEGF及其mRNA的表达.结果 OHSS组和对照组的卵泡液、hCG注射日血清VEGF水平分别为(1 257.2±648.0)与(1 745.1±802.4) ng/L,(250.1±109.5)与(196.7±81.7) ng/L,两组比较,差异无显著性(P>0.05);OHSS组ET日血清VEGF水平及注射hCG后血清VEGF水平变化程度,显著高于对照组[分别为(342.9±158.5)与(222.2±84.6) ng/L,(92.8±106.7)与(25.9±21.8) ng/L,P<0.05].OHSS组取卵日卵巢黄素化颗粒细胞VEGF mRNA的表达,显著高于对照组,VEGF/β肌动蛋白比值分别为1.50±0.60、0.96±0.56(P<0.05).两组取卵日卵巢黄素化颗粒细胞有较活跃的VEGF表达.结论 VEGF在OHSS的发病机理中可能起一定的作用.     

Explorative investigation of vascular endothelial growth factor receptor expression in primary ovarian cancer and its clinical relevance

Objectives

The identification of novel molecular biomarkers, predicting outcome of ovarian cancer, is highly desirable. Considering that angiogenesis is a critical factor for ascites development and peritoneal dissemination in ovarian cancer and given that the vascular endothelial growth factor (VEGF) receptor signaling axis is a major driver of angiogenesis, we sought to analyze expression and compartmental distribution of VEGF-receptor family in ovarian cancer and to assess its clinical relevance with regard to established clinicopathological parameters, tumor cell dissemination to the bone marrow (BM) and the patient's survival.

Methods

A total of 73 patients with primary ovarian cancer were enrolled into this study. Primary tumor tissue was analyzed for the expression of VEGF-R1, VEGF-R2 and VEGF-R3 by immunohistochemistry. The presence of disseminated tumor cells (DTC) in the BM was analyzed by immunocytochemistry using the pancytokeratin antibody A45B/B3 and subsequent automatic detection based on staining and cytomorphology.

Results

In primary ovarian cancer tissue, VEGF-receptor expression, detected with an overall frequency of 44%, was mostly located in the vascular wall and across the stroma; positivity rates for VEGF-R1, VEGF-R2 and VEGF-R3 were 34%, 18% and 26%, respectively. Total VEGF-receptor expression correlated with residual tumor after primary debulking surgery and the presence of DTC at primary diagnosis (p = 0.035, p = 0.023, respectively). Interestingly, VEGF-R1 positivity significantly correlated with decreased progression-free survival (p = 0.026).

Conclusions

This is the first report, suggesting total VEGF-receptor status as a molecular biomarker for monitoring tumor cell spread to the BM and, particularly, revealing prognostic significance of VEGF-R1.     

Expression of cyclooxygenase-2 in epithelial ovarian tumors and its relation to vascular endothelial growth factor and p53 expression

Abstract.   Lee JS, Choi YD, Lee JH, Nam JH, Choi C, Lee MC, Park CS, Juhng SW, Min KW. Expression of cyclooxygenase-2 in epithelial ovarian tumors and its relation to vascular endothelial growth factor and p53 expression. Int J Gynecol Cancer 2006; 16(Suppl. 1): 247–253.
The purpose of this study was to evaluate cyclooxygenase-2 (COX-2) expression in epithelial ovarian tumors and its correlation with vascular endothelial growth factor (VEGF) and p53 expression. Immunohistochemical studies with anti-COX-2, anti-VEGF, and anti-p53 antibodies were carried out in 54 malignant and 23 borderline epithelial ovarian tumors. Elevated COX-2 expression was detected in 77.8% of ovarian carcinomas, which was significantly higher than that of borderline tumors (26.1%) ( P < 0.001). In ovarian carcinomas, there was no significant correlation between COX-2 expression and other clinicopathologic features. Elevated VEGF expression was detected in 74.1% of ovarian carcinomas, and p53 expression was found in 64.8% of ovarian carcinomas. COX-2 expression was statistically correlated with elevated VEGF expression ( P < 0.001) and p53 positivity ( P < 0.05). On a univariate analysis, FIGO stage ( P < 0.0001), histologic type ( P = 0.0104), and COX-2 expression ( P = 0.0135) were significant prognostic factors for overall survival. In a multivariate analysis, FIGO stage ( P < 0.0001) was the only independent prognostic factor for poor survival. These findings suggest that COX-2 may play a role in the progression of epithelial ovarian tumors and that COX-2 expression may contribute to ovarian tumor angiogenesis by stimulating VEGF expression. p53 may be responsible for the regulation of COX-2 expression.     

Prognostic significance of vascular endothelial growth factor expression in ovarian cancer patients: a long-term follow-up

Abstract.   Rudlowski C, Pickart A-K, Fuhljahn C, Friepoertner T, Schlehe B, Biesterfeld S, Schroeder W. Prognostic significance of vascular endothelial growth factor VEGF expression in ovarian cancer patients: a long-term follow-up. Int J Gynecol Cancer 2006; 16(Suppl. 1): 183–189.
The purpose of the study was to determine vascular endothelial growth factor (VEGF) concentrations in ascites from ovarian cancer and to correlate these data with VEGF expression in ovarian tumors, serum VEGF concentrations, and clinicopathologic characteristics. Ascites, serum, and tumor tissue from 65 ovarian carcinomas and eight borderline tumors were collected. VEGF concentration in peritoneal fluids and sera was determined using enzyme immunoassay. VEGF tumor expression was evaluated immunohistochemically. Significantly higher VEGF concentrations were found in ascites from malignant tumors (median, 2575 pg mL⁻¹) compared with borderline tumors (median 181.9 pg mL⁻¹) and benign peritoneal fluid (184.5 pg mL⁻¹). Both VEGF ascites concentration and tumor expression correlated with advanced tumor stages and ascites volume. Elevated VEGF ascites levels were negatively correlated to patient survival. No differences between VEGF serum levels could be observed between ovarian cancer patients and patients with benign cysts. This study showed for the first time the clinical significance of elevated VEGF ascites level in ovarian carcinomas. VEGF is expressed by ovarian tumor cells and locally released in the malignant peritoneal fluid but is not increased in the serum of preoperative ovarian cancer patients. The enhanced VEGF level support novel therapeutic perspectives by VEGF inhibition.     

Serum vascular endothelial growth factor levels before starting gonadotropin treatment in women who have developed moderate forms of ovarian hyperstimulation syndrome.

The study aims to evaluate whether serum vascular endothelial growth factor (VEGF) levels, before treatment with gonadotropins, may be considered a predictive marker of moderate ovarian hyperstimulation syndrome (OHSS). At the University of Pisa hospital infertility unit we have retrospectively selected 10 patients who developed moderate forms of OHSS and 30 control patients who presented a normal response to ovarian stimulation among 400 women undergoing in vitro fertilization (IVF). Serum samples were collected before starting pFSH administration (150-300 IU/day). VEGF levels in serum were measured. No statistically significant difference was found between the serum VEGF levels of patients who developed moderate forms of OHSS and women without any symptoms of the syndrome. Further, serum VEGF concentrations were not significantly correlated with the age of the patients, the number of international units of FSH administered during the cycle of stimulation, the follicle and oocyte numbers counted on the day of the egg retrieval or estradiol levels detected on the same day. This study demonstrates that serum VEGF levels, before starting gonadotropin treatment, are not predictive of the subsequent development of moderate forms of ovarian hyperstimulation syndrome.     

绒毛中表皮生长因子受体和血管内皮生长因子的表达对早孕绒毛血管生成的影响及与稽留流产的关系

目的探讨表皮生长因子受体(EGFR)、血管内皮生长因子(VEGF)的表达对早孕绒毛血管生成的影响及与早孕稽留流产的关系。方法应用免疫组织化学方法检测30例稽留流产(稽留流产组)及30例早孕人工流产(早孕人工流产组)绒毛EGFR、VEGF的表达及绒毛的微血管密度(MVD),并分析相关因子的相关性。结果稽留流产组绒毛的MVD(7.03±1.72)比早孕人工流产组(14.74±1.66)显著降低(P0.01)。稽留流产组绒毛的EGFR表达水平(0.42±0.14)表达比早孕人工流产组(0.49±0.07)低(P0.05),稽留流产组绒毛的VEGF表达水平(0.47±0.12)表达比早孕人工流产组(0.60±0.12)低(P0.01)。Pearson相关分析显示在早孕人工流产绒毛及稽留流产绒毛中EGFR与VEGF的表达呈正相关,EGFR的表达与MVD呈正相关,VEGF表达与MVD呈正相关。结论绒毛组织中EGFR和VEGF的表达与绒毛血管生成状况密切相关,EGFR和VEGF低表达参与了稽留流产的绒毛血管的形成障碍。     

The expression of epidermal growth factor receptor, vascular endothelial growth factor, matrix metalloproteinase-2, and cyclooxygenase-2 in relation to human papilloma viral load and persistence of human papillomavirus after conization with negative margins

The aim of this study was to investigate the correlations between human papillomavirus (HPV) load and vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), matrix metalloproteinase-2 (MMP-2), and cyclooxygenase-2 (COX-2), and to identify biomarkers that may predict high-risk HPV clearance or persistence after conization with negative margins. The following samples were analyzed: 77 paraffin-embedded specimens from patients with cervical intraepithelial neoplasia (CIN), including 27 CIN 2 conization specimens and 50 CIN 3 conization specimens. Immunohistochemical analysis was performed with antibodies to VEGF, EGFR, MMP-2, and COX-2. Hybrid capture II testing was used to detect HPV DNA. VEGF expression was significantly associated with HPV load (rho = 0.27186, P = 0.0191), while COX-2 expression was significantly and inversely associated with HPV load (rho = -0.34309, P = 0.0028). In univariate analysis, HPV load (P = 0.0112) and VEGF expression (P = 0.0274) were significantly associated with high-risk HPV clearance or persistence after conization with negative margins. In multiple regression analysis, high viral load (relative light unit/positive control > 500) and positive VEGF expression were significantly associated with high-risk HPV persistence after conization with negative margins (odds ratio [OR]: 9.915, CI: 1.891-51.994; OR: 6.661, CI: 1.208-36.722, respectively). In conclusion, VEGF expression is related to HPV load, while COX-2 expression is inversely related to HPV load, and immunohistochemical analysis of VEGF expression and HPV viral load are a significant and an independent prognostic indicator of high-risk HPV persistence after conization with negative margins.     

Elevated level of plasma vascular endothelial growth factor after gonadotropin-releasing hormone agonist treatment for leiomyomata

Objective. Uterine leiomyomata are the most common gynecological benign tumor and greatly affect reproductive health and well-being. The pathophysiology and epidemiology of fibroids are poorly understood. Gonadotropin-releasing hormone agonist (GnRH-a) pretreatment is one of the unfavourable factors for leiomyomata treatment with uterine artery embolisation (UAE). In this study, we investigated the plasma level of vascular endothelial growth factor (VEGF) in uterine leiomyoma patients with or without GnRH-a pretreatment.

Study design. Thirty-two women who underwent UAE for symptomatic uterine leiomyoma were analysed. The plasma level of VEGF was studied before UAE.

Results. The level of plasma VEGF was significantly higher in the GnRH-a pretreated group compared with the non-treated group.

Conclusion. A compensative reaction for vasculature after GnRH-a treatment is speculated. Higher level of VEGF in GnRH-a pretreatment group could be one of the unfavourable factors for the treatment of uterine leiomyomata by UAE.