Vascular Endothelial Growth Factor In Ascites Research Articles

Angio-Suppressive Effect of Partially Purified Lectin-like Protein from Musa acuminata pseudostem by Inhibition of VEGF-Mediated Neovascularization and Induces Apoptosis Both In Vitro and In Vivo

Lifestyle and nutritional changes have contributed much to the somatic genetic changes which have concurrently led to an increase cancer in humans. Hence the plant-based and nutritional involvements block oncogenic transformation are in good demand. We evaluate Phloem exudates of the dietary plant, Musa acuminate pseudostem, the initial domesticated plant species with the effective lectin activity for its functional role against the tumor development and its mechanism of action. Our experimental data exhibit that Musa acuminata Lectin Protein (MALP) shows a promising cytotoxic effect against the various human cancer cell lines. Supporting this, we evaluate the in vivo anti-tumor and anti-angiogenic activity of MALP in Ehrlich Ascites Carcinoma mice model (EAC). MALP treatment resulted in tumor growth inhibition and increased the lifespan of the EAC-bearing mice without showing any side effects on normal mice, as revealed by histological parameters. Further, a significant decrease in the ascites vascular endothelial growth factor (VEGF) secretion and microvessel density supports the anti-angiogenic property of the MALP. Apoptosis-inducing activity of MALP was revealed by DNA fragmentation assay, Caspase-3 inhibitor assay and cellular morphology were studied by fluorescence staining methods. Our study delivers the real evidence that MALP with a promising an anticancer potential expressively degenerates the tumor development by affecting angiogenesis and apoptosis.

LQFM030 reduced Ehrlich ascites tumor cell proliferation and VEGF levels

AimsThis study reports the biological properties of LQFM030 in vivo, a molecular simplification of the compound nutlin-1. Main methodsEhrlich ascites tumor (EAT)-bearing mice were treated intraperitoneally with LQFM030 (50, 75 or 150mg/kg) for 10days to determine changes in ascites tumor volume, body weight, cytotoxicity and angiogenesis. Moreover, flow cytometric expression of p53 and p21 proteins and caspase-3/7, −8 and −9 activation were investigated in EAT cells from mice treated. Acute oral systemic toxicity potential of LQFM030 in mice was also investigated using an alternative method. Key findingsTreatment of EAT-bearing mice with LQFM030 resulted in a marked decline in tumor cell proliferation and the vascular endothelial growth factor (VEGF) levels along with enhanced survival of the mice. Apoptotic tumor cell death was detected through p53 and p21 modulation and increase of caspase-3/7, −8 and −9 activity. LQFM030 also showed orally well tolerated, being classified in the UN GHS category 5 (LD50>2000–5000mg/Kg). SignificanceLQFM030 seems to be a promising antitumor candidate for combinatory therapy with typical cytotoxic compounds, reducing the toxicity burden while allowing a superior anticancer activity. Moreover, these data also open new perspectives for LQFM030 as an antiangiogenic agent for treatment of diseases involving VEGF overexpression.

A Novel Combination of C-Reactive Protein and Vascular Endothelial Growth Factor in Differential Diagnosis of Ascites.

Ascites with unknown cause remains a diagnostic challenge, which needs novel noninvasive biomarkers for the precise diagnosis. We aimed to evaluate the ascitic fluid and serum C-reactive protein (CRP) and vascular endothelial growth factor (VEGF) as diagnostic markers in the differential diagnosis of malignant and benign ascites. In this prospective work, 315 consecutive patients with ascites were studied. Ascitic fluid and serum levels of CRP and VEGF were evaluated by using an enzyme-linked immunosorbent assay. Patients were divided into a benign ascites group (group 1) (n=256) and a malignant ascites group (group 2) (n=59). Ascitic and serum CRP were significantly elevated in malignant ascites than benign ascites group [5.08 (3.62-6.58) vs. 1.82 (0.64-3.86) ng/ml; P<0.001 and 12.7 (8.55-17.05) vs. 5.94 (2.57-10.64) ng/ml; P<0.001], respectively. Ascitic and serum VEGF were significantly increased in malignant ascites than benign ascites patients [0.68 (0.39-0.96) vs. 0.41 (0.25-0.83) ng/ml; P<0.001 and 0.74 (0.41-1.08) vs. 0.54 (0.23-0.86) ng/ml; P<0.001], respectively. At a cutoff value of 7.3 and 0.63ng/ml, serum CRP and VEGF had specificity (77.3 and 89.5%) and sensitivity (83.1 and 94.9%) for detecting malignant ascites [area under the curve (AUC) 0.821, 0.921], respectively. At a cutoff value of 2.5 and 0.57ng/ml, ascitic CRP and VEGF had specificity (81.6 and 85.5%) and sensitivity (84.7 and 91.5%) for detecting malignant ascites (AUC 0.842, 0.894), respectively. Elevated ascitic fluid and serum CRP and VEGF values were related to the malignant ascites.

Does VEGF facilitate local tumor growth and spread into the abdominal cavity by suppressing endothelial cell adhesion, thus increasing vascular peritoneal permeability followed by ascites production in ovarian cancer?

BackgroundOvarian cancer is mostly associated with pathologically regulated permeability of peritoneal vessels, leading to ascites. Here, we investigated the molecular regulation of endothelial permeability by the vascular endothelial growth factor (VEGF) and both tight and adherens junction proteins (VE-cadherin and claudin 5) with regards to the tumor biology of different ovarian cancer types.MethodsSerum and ascites samples before and after surgery, as well as peritoneal biopsies of 68 ovarian cancer patients and 20 healthy controls were collected. In serum and ascites VEGF protein was measured by ELISA. In peritoneal biopsies co-localization of VE-cadherin and claudin 5 was investigated using immunohistochemical dual staining. In addition, the gene expression of VE-cadherin and claudin 5 was quantified by Real-time PCR. Differences in VEGF levels, VE-cadherin and claudin 5 gene expression were analyzed in relation to various tumor characteristics (tumor stage, grading, histological subtypes, resection status after surgery) and then compared to controls. Furthermore, human primary ovarian cancer cells were co-cultured with human umbilical vein endothelial cells (HUVEC) and changes in VE-cadherin and claudin 5 were investigated after VEGF inhibition.ResultsVEGF was significantly increased in tumor patients in comparison to controls and accumulates in ascites. The highest VEGF levels were found in patients diagnosed with advanced tumor stages, with tumors of poor differentiation, or in the group of solid / cystic-solid tumors. Patients with residual tumor after operation showed significantly higher levels of VEGF both before and after surgery as compared to tumor-free resected patients. Results of an immunohistochemical double-staining experiment indicated co-localization of VE-cadherin and claudin 5 in the peritoneal vasculature. Compared to controls, expression of VE-cadherin and claudin 5 was significantly suppressed in peritoneal vessels of tumor patients, but there were no significant differences regarding VE-cadherin and claudin 5 expression in relation to different tumor characteristics. A significant positive correlation was found between VE-cadherin and claudin 5 expression. VEGF inhibition in vitro was associated with significant increase in VE-cadherin and claudin 5.ConclusionsOur results indicate that increased peritoneal permeability in ovarian cancer is due to down-regulation of adhesion proteins via tumor derived VEGF. Advanced ovarian cancer with aggressive tumor biology may be associated with early dysregulation of vascular permeability leading to ascites. These patients may benefit from therapeutic VEGF inhibition.

The clinical significance of vascular endothelial growth factor in malignant ascites

Ascites can be caused by many kinds of diseases. Patients with undetermined ascites represent a diagnostic challenge. The aims of this study were to determine the diagnostic value of vascular endothelial growth factor (VEGF) in differentiation of malignant ascites from benign ascites and to investigate the clinical value of ascitic VEGF as an independent prognostic parameter. The study included 462 consecutive patients with malignant ascites and 550 patients with benign ascites, VEGF level in ascites were determined by a sandwich enzyme-linked immunosorbent assay. The survival rate was calculated by the Kaplan-Meier method and the log-rank test. Multivariate survival analysis was performed using the Cox hazards model. In our study, we found VEGF levels in malignant ascites (676.59 ± 303.86pg/ml) were significantly higher than those in benign ascites (218.37 ± 98.15pg/ml) (P < 0.001). Meanwhile, we also found that VEGF levels in malignant ascites from patients with ovarian cancer were higher than those with other cancers. Areas under the receiver operating characteristic (ROC) curves of ascitic VEGF was 0.940. At a cutoff value of 319.5pg/ml, VEGF yielded a sensitivity of 89.2% and a specificity of 88.4%. Patients associated with the high-level VEGF value (≥613.38pg/ml) in malignant ascites exhibited poor mean survival rates (8.3 ± 0.52 vs 15.11 ± 0.66months, P < 0.001). In a multivariate Cox regression model, higher ascitic VEGF was an independent prognostic factor for overall survival. Planned subgroup analysis was performed for patients with tumor node metastasis (TNM) stage I. In the univariate analysis, only ascitic VEGF was associated with overall survival. VEGF was found to have a highly accurate sensitivity and specificity, suggesting that it could be considered as a new biomarker to differentiate malignant ascites from the benign one. The high level of VEGF value in malignant ascites may be used as an independent prognostic factor in patients with all stages of cancer.

Intraperitoneal Bevacizumab (Bev) for Control of Malignant Ascites Due to Advanced-Stage Gastrointestinal Cancers (Gi): a Multicenter Double-Blind, Placebo (Pl)-Controlled, Randomized Phase Ii Study. Aio Sup-0108

ABSTRACT Aim: Malignant Ascites is debilitating for pts. with advanced cancer which negatively impacts the quality of life (QoL). Causative treatment approaches are still limited. It has previously been shown that tumor cell production and/or increases of Vascular Endothelial Growth Factor (VEGF) might be a major cause of the formation of malignant ascites. Intraperitoneal Bev could therefore be an option for symptom control of refractory malignant ascites. Methods: Pts. with advanced GI cancer and malignant ascites who had received paracentesis at least once within the past 4 weeks were randomly assigned in a 2:1 ratio to intraperitoneal Bev (400mg absolute in 100 ml NaCl 0.9%) or Pl after paracentesis. During the 8-week treatment period, a minimum interval of 14d was kept between the applications of the study drug. First primary endpoint was the paracentesis-free survival (ParFS). Second primary endpoint was best response (BR) defined as the longest paracentesis-free period within the 12-week observation period. Further endpoints were overall survival, QoL, serum and ascites VEGF. ParFS and BR were compared using the logrank test and the Wilcoxon-Mann-Whitney test, respectively. Results: 53 Patients (median age 63y) were randomized (37 Bev/16 Pl arm) whereas 49 pts. received at least one application of the study drug and qualified for the intention to treat analysis. The median ParFS was 14d (CI:11-17d) in the Bev arm and 10.5d (CI: 7-21d) in the Pl arm (hazard ratio 0.74, CI:0.40-1.37; p = 0.16). The BR was 19d for the Bev arm (range 6-66d) and 17.5d for the Pl arm (range 4-42d) with a p value of 0.85. Median OS was 64d (CI: 45-103d) for the Bev arm and only 31.5d (CI: 20-117d) for the Pl arm (p = 0.31). The proportion of pts. with at least one CTC grade 3-5 event occurred was similar with 20/33 (61%) in the Bev arm and 11/16 (69%) in the Pl arm. Results on serum and ascites VEGF will be presented at the meeting. Conclusions: In this unfavorable group of terminally ill pts. intraperitoneal Bev was well tolerated but did not result in a significantly better symptom control of malignant ascites compared to the control. Disclosure: D. Atanackovic: Research Support Roche; D. Arnold: Honoraria & Research Support: Roche; J. Russel: Advisory Board Roche. All other authors have declared no conflicts of interest.

Effect of gold nanoparticles on expression and secretion of vascular endothelial growth factor by HepG2 cells

Objective To observe the inhibitory effect of gold nanoparticles on the expression and secretion of vascular endothelial growth factor (VEGF) by HepG2 cells.Methods HepG2 cells were seeded in 96-well plates and treated with different concentrations (5,10,and 20 μg/L) of gold nanoparticles for 12 h.The concentration of VEGF protein expressed and secreted by HepG2 cells was determined by using Western blotting and enzyme linked immunosorbent assay (ELISA) separately.The proliferation rate of HepG2 cells was assessed by using methyl thiazol tetrazolium (MTT) assay.For mouse ascites model experiments,H22 cells were intraperitoneally injected into BALB/c mice.After 7 days,gold nanoparticles (1 mg/kg) was administered through intraperitoneal injection every 2 days over a period of 14 days.Mice were then sacrificed and ascites fluids collected.The fluid accumulation in the peritoneal carvity,as well as the concentration of VEGF of mouse ascites was determined.Results The VEGF concentration of 4 groups (control,5,10 and 20 μg/L gold nanoparticles) was (351.64 ±7.89),(285.62 ± 3.45),(121.72 ±3.10) and (9.83 ±2.28) ng/L respectively,P ＜0.05.The proliferation rate of HepG2 cells in three gold nanoparticles group (5,10,and 20 μg/L) was (98.98 ±7.57)％,(90.95 ± 11.62)％ and (86.38 ±4.23)％ respectively,P ＞ 0.05.The ascites accumulation and VEGF concentration in control and gold nanoparticles groups were (13.22 ± 1.03) ml and (62.95 ± 11.93) ng/L,and (5.21 ± 0.62) ml and (27.12 ± 8.58) ng/L respectively,P ＜ 0.01.Conclusion Gold nanoparticles can inhibit the expression and secretion of VEGF and cause lesser ascites accumulation of mouse ascites model. Key words: Carcinoma, hepatocellular; Gold nanoparticles; Vascular endothelial growth factor; Ascites accumulation

VEGF is a target molecule for peritoneal metastasis and malignant ascites in gastric cancer: prognostic significance of VEGF in ascites and efficacy of anti-VEGF monoclonal antibody

BackgroundIn gastric cancer, poor prognosis is associated with peritoneal dissemination, which often accompanies malignant ascites. We searched for a target molecule in peritoneal metastasis and investigated its clinical utility as a biomarker.MethodsBiopsy specimens from both primary lesions and peritoneal metastasis, and if possible, malignant ascites, were obtained from 40 patients with gastric cancer. Vascular endothelial growth factor (VEGF) expression was analyzed by immunohistochemical staining and enzyme-linked immunosorbent assay.ResultsVEGF expression was seen in 70% of peritoneal samples. Of the 40 patients, 35 had malignant ascites. These 35 patients were divided into two groups: 15 with ascites found beyond the pelvic cavity (large group) and 20 whose ascites were within the pelvic cavity (small group). The two groups did not significantly differ by serum VEGF levels, but ascites VEGF levels in the large group were significantly higher than in the small group (P < 0.0001). Serum VEGF and ascites VEGF levels were highly correlated in the large group (r = 0.686). A high ascites VEGF level was found to be a risk factor for survival (P = 0.045). We include a report of a patient with chemoresistant refractory gastric cancer and symptomatic ascites who obtained 8 months of palliation from systemic bevacizumab.ConclusionAnti-VEGF therapies are promising, and the ascites VEGF level is an important marker in managing patients with gastric cancer and peritoneal metastasis.

Clinical study of intraperitoneal injection bevacizumab (BV) combined with intraperitoneal hyperthermic perfusion chemotherapy (CT) in treatment of malignant ascites of ovarian cancer (OC).

5558 Background: To study the efficacy and safety of intraperitoneal injection BV combined with intraperitoneal perfusion CT in treatment of malignant ascites of OC and to investigate the clinical significance of the concentration change of vascular endothelial growth factor (VEGF). Methods: Patients with malignant ascites of OC were randomly divided into treatment group (n = 31) and control group (n = 27). All enrolled patients received TC (paclitaxel 135mg/m2, iv d1+ carboplatin AUC=5, iv d1), 1 time/3 weeks for 6 weeks. Patients in the control group were treated with intraperitoneal perfusion CT combined with intraperitoneal cisplatin (40mg/m2)1 time/2 weeks for 6 weeks. Besides the treatment of the control group, patients in the treatment group received intraperitoneal injection with BV 300mg after each intraperitoneal perfusion CT for 6 weeks. The improvement of life quality, efficacy and adverse effects were evaluated. The concentrations of VEGF and CA-125 in ascites of the two groups were assayed by ELISA method. Results: No severe side effect was observed in all the patients. The response rate of the treatment group was higher than that of the control group (90.32% vs. 59.26%, p &lt; 0.05). The improvement rate of quality of life (QOL) of the patients in the treatment group was also higher than that of the patients in the control group (93.55 % vs. 48.15%, p &lt; 0.05). The concentration of VEGF in ascites of the treatment group after treatment was lower than that before treatment (p &lt; 0.05). And after treatment, the concentration of VEGF in ascites of treatment group was lower than that of the control group (p &lt; 0.05). Conclusions: Intraperitoneal injection BV combined with intraperitoneal perfusion CT in the treatment of malignant ascites of OC is effective and safe. The concentration of VEGF and CA-125 levels, in ascites may be used as monitoring index for treatment effect of malignant ascites of OC with intraperitoneal injection BV.

Differential Effect of Intraperitoneal Albendazole and Paclitaxel on Ascites Formation and Expression of Vascular Endothelial Growth Factor in Ovarian Cancer Cell-Bearing Athymic Nude Mice

The purposes of our study were to evaluate the effect of intraperitoneal albendazole on tumor growth, ascites formation, and vascular endothelial growth factor (VEGF) mRNA expression, and to assess the synergistic effect of paclitaxel in OVCAR-3-bearing nude mice. In all, 4 groups of mice were injected intraperitoneally with weekly albendazole (450 mg/kg per week), paclitaxel (30 mg/kg per week), albendazole plus paclitaxel, or normal saline for 4 weeks. Ascitic fluid accumulation (2.47, 2.65, 2.88, and 5.90 mL, respectively) and in ascitic VEGF levels were significantly reduced in the 3 treatment groups compared to the control group (170.83, 229.16, 267, and 1625 pg/mL, respectively). However, complete tumor suppression was more prominent in the paclitaxel group, and VEGF mRNA expression was more strongly inhibited in the albendazole group (P < .05). No synergistic effect of albendazole and paclitaxel was observed. We demonstrated a differential effect of albendazole and paclitaxel in a xenograft model of ovarian carcinoma; albendazole suppressed ascites formation by inhibiting VEGF secretion, and paclitaxel exerted its effects by direct cytotoxicity.

The effect of VEGF on T cells from ovarian cancer patients and healthy individuals via VEGF receptor type 2.

e15552 Background: The role of vascular endothelial growth factor (VEGF) in tumor angiogenesis is well characterized; nevertheless, it is also a key element in promoting tumor evasion of the immune system by down-regulating dendritic cell maturation and thus T cell activation. We investigated the possible direct effect of VEGF on T cell activation and through which type of VEGF receptor (VEGFR) it exerts this effect. Methods: Circulating T cells from healthy donors (n=9) and ovarian cancer patients (n=5) were expanded in cultures with anti-CD3 and IL-2 with or without VEGF for 14 days, and the number of T cells was assessed. Cultured T cells were also tested for their cytotoxic activity against K562 and Daudi cell lines, in a standard 4-h 51Cr-release assay, and the expression of VEGFRs 1, 2, and 3 was assayed by flow cytometry, immunocytochemistry, and Western blotting. To assess the ability of activated T cells to secrete VEGF, levels in culture supernatants were measured by ELISA. Results: The addition of VEGF in cultures showed that concentrations above 1 ng/ml significantly reduced T cell proliferation (p=0.043 for comaprison with controls). The suppressive effect was dose-dependent from 1 to 10 ng/ml, after which there was a plateau of the inhibitory effect. Protein expression studies demonstrated that CD3+ T cells express VEGFR-2 on their surface upon activation. The direct suppressive effect of VEGF on T cell proliferation was reversed by anti-VEGFR-2 antibodies. This finding proves that the suppressive effect of VEGF on T cells is mediated by VEGFR-2. We also showed that VEGF at 5 ng/ml significantly reduced the cytotoxic activity of T cells against K562 and Daudi cells (p=0.042 for comparison with controls). Activated T cells secreted VEGF in the culture environment (mean value: 192 pg/ml). Nevertheless, these levels are well below those necessary to produce T cell suppression in culture (1 ng/ml). Conclusions: This study shows that VEGF directly suppresses T cell activation via VEGF receptor type 2. The levels shown to produce suppression in vitro (>= 1 ng/ml) are in concert to the level of VEGF in ascites, which predicted for a poor outcome (1.9 ng/ml) in a previous study (Bamias et al, Gynecol Oncol 2008).

Study of T Lymphocytes Infiltrating Peritoneal Metastases in Advanced Ovarian Cancer

The presence of CD3(+) tumor-infiltrating lymphocytes (TILs) has been found to correlate with improved survival in epithelial ovarian cancer, but the association of TIL subpopulations with clinical outcome remains controversial. We performed a prospective analysis of TIL subpopulations from patients with epithelial ovarian cancer and their activation status and studied their association with prognosis. Flow cytometric analysis was performed on TIL subpopulations isolated from 45 fresh ovarian tumor specimens, obtained during surgery, after mechanical dissociation and enzymatic degradation. Vascular endothelial growth factor and tumor necrosis factor alpha levels in ascites and serum were measured by enzyme-linked immunosorbent assay. Significantly increased numbers of CD56(+) cells (natural killer and natural killer-like T cells; P = 0.045), activated CD4(+)HLA-DR cells (P = 0.046), and activated CD8(+)CD25(+) cells (P = 0.028) were found in serous and endometrioid carcinomas compared with mucinous and clear cell carcinomas. A high percentage of CD4(+)CD25(hi) cells (regulatory T cells) and activated CD4(+)HLA-DR cells significantly associated with improved median overall survival (not reached vs 35 months [P = 0.0241] and not reached vs 35 months [P = 0.0144], respectively) and median progression-free survival (30 months vs 14 months [P = 0.0819] and 30 months vs 13 months [P = 0.0479], respectively). Vascular endothelial growth factor ascites levels were inversely correlated with CD14(+) (rho = -0.529, P = 0.001), whereas HLA-DR8(+)CD8 lymphocytes were inversely correlated with both ascites and serum vascular endothelial growth factor levels (rho = -0.494, P = 0.006, and rho = -0.586, P = 0.037, respectively). The presence of regulatory T cells and activated CD4(+) cells within the tumor microenvironment is associated with improved overall and progression-free survival in patients with ovarian cancer.

Vascular endothelial growth factor (VEGF) is associated with reduced NK-like T cells in ascites and suppresses lymphocytes in cell cultures: Prognostic implications of VEGF in ascites from ovarian cancer

16505 Background: We have shown that CD4+CD25+ Tregs and CD3+CD56+ NK-like T cells are significantly increased in the ascites from ovarian cancer compared with ascites from cirrhosis or blood from the same patients (Gynecol Oncol 2007; 106: 75–81, 2007). Both populations are involved in the immune response against the tumor. Since VEGF may be involved in immune regulation, we studied its association with these populations and prognosis. We also studied this relationship by ex vivo expansion of these cells in culture. Methods: 64 patients with ovarian cancer were included. VEGF, TNFa, IFNγ, IL-10 and IL-12 in ascites and serum were measured by ELISA, while cell populations and expression of VEGF receptors were studied with flow cytometry. For tissue culture experiments, 6 × 106 cells were plated on Petri dishes and were pulsed with IL-2, anti-CD3 and IFN-γ in order to expand CD3+ and CD56+ cells. Blood from healthy individuals was also analysed. VEGF at 0, 20, 100 and 500 pg/ml was also added to the cultures. Each concentration was tested in triplicate. Results: VEGF ascites levels were inversely correlated with CD3+CD56+ cells (rho= -0.316, p=0.012). Among patients receiving first-line chemotherapy (n=42), VEGF levels ≤ 1,900 pg/ml were associated with platinum sensitivity (p=0.030), improved progression-free survival (17 vs. 9 months, p=0.028) and overall survival (not reached vs. 20 months, p=0.045). Cell culture experiments showed that VEGF resulted in reduction of CD3+ and CD56+ cells from blood and ascites from cancer patients (Table 1). The expression of VEGF receptors on these cells was low and it was not increased by VEGF. Conclusions: VEGF ascites levels are associated with prognosis in advanced ovarian cancer. Their prognostic significance may be due to the suppression of NK T-like CD3+CD56+ cells. The mechanism of this suppresison remains unclear and it is not through the induction of VEGF receptors on these cells. Total cell counts (×106) in cell cultures with 0, 20, 100 and 500 pg/ml of VEGF Days VEGF 0 pg/ml VEGF 20 pg/ml VEGF 100 pg/ml VEGF 500 pg/ml Healthy (n=3) 0 6 6 6 6 Healthy 7 26.2 24 18 20 Healthy 14 131 122 74.9 106 OvCa blood (n=4) 0 6 6 6 6 OvCa blood 7 10.7 10.8 8 9.2 OvCa blood 14 116 61 66 58 OvCa ascites (n=4) 0 6 6 6 6 OvCa ascites 7 20 12 12 11.5 OvCa ascites 14 150 80 77 60 OvCa: ovarian cancer No significant financial relationships to disclose.

Correlation of NK T-like CD3+CD56+ cells and CD4+CD25+(hi) regulatory T cells with VEGF and TNFα in ascites from advanced ovarian cancer: Association with platinum resistance and prognosis in patients receiving first-line, platinum-based chemotherapy

Background Several cytokines have been associated with immune regulation and prognosis in ovarian cancer. CD4+CD25+ Tregs and CD3+CD56+ NK-like T cells are involved in the immune response against the tumor. We have investigated the association of cytokines in the ascites from patients with ovarian cancer with these populations, the platinum resistance and the prognosis of these patients. Patients and methods Ascites from 64 patients with ovarian cancer was analysed. Forty-two patients were studied at diagnosis (FIGO stage III in 40 cases) and were treated with cytoreductive surgery and platinum-based chemotherapy. Ascites from 9 patients with cirrhosis was used as control. Vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNFα), interferon-gamma (IFNγ), inteleukin-10 (IL10) and interleukin-12 (IL12) in ascites and serum were measured by enzyme-linked immunosorbent assay (ELISA), while lymphocytic populations were studied with three-colour flow cytometry. Results VEGF ascites levels were inversely correlated with CD3+CD56+ cells (rho = − 0.316, p = 0.012), while a similar correlation was observed between TNFα ascites levels and CD4+CD25+(hi) cells (rho = − 0.332, p = 0.041). Among patients receiving first-line chemotherapy, VEGF levels < 1900 pg/ml and TNFα levels > 35 pg/ml were associated with platinum sensitivity ( p = 0.021 and p = 0.028, respectively) and improved progression-free survival (PFS) ( p = 0.007 and p = 0.045, respectively). Low VEGF levels were also associated with improved overall survival ( p = 0.026). Conclusion VEGF and TNFα ascites levels are associated with prognosis in advanced ovarian cancer. Their prognostic significance may be due to their association with immunologically important populations, namely the NK T-like CD3+CD56+ cells and the Tregs CD4+CD25+(hi) cells.

Intraperitoneal gene therapy by rAAV provides long-term survival against epithelial ovarian cancer independently of survivin pathway

Epithelial ovarian carcinoma is the leading cause of death from gynecological malignancies. Owing to the lack of an effective screening method, insidious onset, and non-specific symptoms, a majority of women present with advanced stage disease. Despite improvements from cytoreductive surgery and chemotherapy, recurrent disease remains a formidable challenge. In the present study, we demonstrate for the first time that stable intra-abdominal genetic transfer of endostatin and angiostatin (E+A) by recombinant adeno-associated virus (rAAV) provides sustained antitumor effects on the growth and dissemination of epithelial ovarian cancer in a mouse model. Further, when combined with paclitaxel (taxol), the effect of this therapy was dramatically increased and resulted in long-term tumor-free survival overcoming prior limitations of chemotherapy and gene therapy. The combined effects of angiosuppressive therapy and chemotherapy were found to be independently of survivin pathway. Evidence for the superior effects of the combination therapy was indicated by significantly lower ascites volume with less hemorrhage and tumor conglomerates, lower ascites vascular endothelial growth factor, higher tumor cell apoptosis and decreased blood vasculature, and long-term disease-free survival. Histopathology of visceral organs and liver enzyme assays indicated no toxicity or pathology.

Albendazole: a Potent Inhibitor of Vascular Endothelial Growth Factor and Malignant Ascites Formation in OVCAR-3 Tumor-Bearing Nude Mice

Angiogenesis and vessel hyperpermeability are the two factors leading to the formation of ascites. Vascular endothelial growth factor (VEGF) plays a pivotal role in malignant ascites formation. We have recently shown that albendazole inhibits peritoneal growth of human colorectal cancer cells (HT-29). The present study was designed to find out if albendazole can suppress ascites formation in ascites-producing peritoneal carcinomatosis. Female nude mice bearing peritoneal tumors of human ovarian cancer cells (OVCAR-3) were treated with albendazole. Following i.p. inoculation and ascites development, mice were given i.p. albendazole (150 mg/kg) or the vehicle x 3 weekly for 4 weeks. Whereas vehicle-treated mice developed overt ascites requiring repeated aspiration, ascites formation in the albendazole-treated mice was markedly suppressed. As a result of this, 7 of 10 mice from the control group had to be euthanized before the course of treatment was over. Suppressed ascites production and reduced tumor vascularity observed was a result of dramatic reduction in tumor VEGF production as revealed by profoundly lower VEGF ascites fluid and plasma levels. In vitro, incubation of SKOV-3 cells with various concentrations of albendazole led to significant dose-dependent inhibition of VEGF secretion. Examination of floating tumor cells collected from the peritoneal wash revealed profound down-regulation of VEGF mRNA in albendazole-treated mice. These findings suggest for the first time that in nude mice bearing OVCAR-3 peritoneal tumors, by inhibiting VEGF production, albendazole abolishes tumor angiogenesis and ascites formation.

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

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.

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

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.

Phosphatidylinositol 3-Kinase Inhibitors Are a Triple Threat to Ovarian Cancer

In this issue of Clinical Cancer Research, Hu et al. assess the role of phosphatidylinositol 3-kinase (PI3K) inhibition in vascular permeability, angiogenesis, and vascular remodeling in both tumor vessels and the peritoneal lining an athymic mouse model of intraperitoneal human ovarian carcinoma ([

Lysophosphatidic Acid (LPA)-Induced Vascular Endothelial Growth Factor (VEGF) by Mesothelial Cells and Quantification of Host-Derived VEGF in Malignant Ascites

Lysophosphatidic acid (LPA) is a lipid mediator with multiple biological activities that may affect the progression of various cancers. Malignant ascites contains high levels of LPA as well as vascular endothelial growth factor (VEGF). Although LPA receptors are widely expressed in normal as well as cancer cells, little is known about the effect of LPA on host cells. Therefore, we evaluated the effect of LPA specifically on peritoneal mesothelial cells (PMC), and assessed another aspect of LPA in tumor biology mediated through the host cells. The effect of LPA on the production of VEGF was evaluated by ELISA and northern blotting. Next, we quantified human- and mouse-VEGF separately in ascitic fluid of nude mice inoculated intraperitoneally with a human gastric cancer, MKN45, and thus evaluated the ratio of host-derived VEGF in malignant ascites. Addition of 10 to 80 mum LPA enhanced VEGF production by PMC through gene activation. The effect was strongly inhibited by pre-treatment with PTX or Ki16425, indicating that the effect was mainly dependent on the LPA1 signal. Of the VEGF in ascitic fluid at 3 weeks after tumor inoculation, 12.8% was derived from mouse cells. At 6 weeks, however, the ratio of host-derived VEGF was reduced to 5.0%, suggesting that the ratio of host-derived VEGF may be higher in the earlier phase. Because tumor growth is often associated with an increase of LPA concentration in ascites, stimulation of VEGF production in PMC might have an important role in the growth of cancer cells disseminated in the peritoneal cavity.