Endostar Group Research Articles

Endostar combined with cryoablation for subcutaneous xenografted tumor model of lung adenocarcinoma cell line A549 in BALB/c nude mice: An experimental study

Objective To investigate the inhibitory effect of Endostar combined with cryoablation on lung adenocarcinoma cell line A549 in BALB/c nude mice, and to discuss its interaction mechanisms.Methods The lung adenocarcinoma A549 model in BALB/c nude mice were established. When the largest diameter of tumor reached 1.0 cm, a total of 24 mice were randomly and equally divided into 4 groups:control group, Endostar group, cryoablation group and cryoablation plus Endostar group. The largest diameter and the vertical diameter of the tumors were measured at different points of time after treatment. At the 21st day, the mice were sacrificed and the tumors were removed and the rate of tumor cell apoptosis, the microvessel density(MVD) and the expression level of vascular endothelial growth factor(VEGF) were determined by using immunohistochemistry method. The results were statistically analyzed. Results The tumor growth velocity of the control group, Endostar group, cryoablation group and cryoablation plus Endostar group was(236.68 ± 51.23)%,(220.02 ± 30.61)%,(159.46 ± 29.33)% and(103.34 ± 25.50)%,respectively(P 0.01). The rate of apoptosis of the four groups was(21.67 ± 2.34)%,(22.17 ± 1.47)%,(38.33 ± 1.37)% and(49.17 ± 1.72)%,respectively(P 0.01). The MVD and the expression levels of VEGF of the cryoablation plus Endostar group were significantly lower than those of the other three groups(P 0.01). Statistical analysis revealed that a positive correlation existed between the express of VEGF and MVD. Conclusion Endostar can obviously enhance the therapeutic efficacy of cryoablation on lung adenocarcinoma A549 in BALB/c nude mice. The underlying mechanisms may be the Endostar-inhibited angiogenesis through down-regulating the expression of VEGF, and the cooperative effect of Endostar and cryoablation on the promotion of tumor cell apoptosis.

Suppression of choroidal neovascularization by Endostar in rats.

Choroidal neovascularization (CNV) is common in various retinal and choroidal diseases, and may result in severe and irreversible loss of vision. Our previous studies suggested that Endostar, a novel recombinant endostatin, is able to inhibit the proliferation and migration of choroid‑retinal endothelial cells. To further evaluate the effect of Endostar on the formation of CNV in vivo, a rat model of laser‑induced CNV was constructed and Endostar or phosphate‑buffered saline treatment was administered intravitreally every other day. Using fluorescein angiography (FA), reduced CNV incidence and leakage grade was observed in the Endostar group. In addition, CNV area and maximal thickness were prominently reduced in the Endostar group measured by choroid flat mounts and sections. Furthermore, vascular endothelial growth factor (VEGF), hypoxia‑inducible factor 1α and chemokine C‑X‑C motif ligand 1 were markedly reduced in the Endostar group as determined by quantitative polymerase chain reaction and downregulation of VEGF was also verified by western blot analysis at the protein level. This study demonstrates that Endostar suppressed CNV in a rat model, which may be largely mediated by the downregulation of VEGF and other angiogenic molecules.

Inhibitory effect of endostar on lymphangiogenesis in non-small cell lung cancer and its effect on circulating tumor cells

背景与目的血管内皮抑素可以抑制肿瘤新生血管的生成，但对肿瘤微淋巴管的形成与发展是否存在抑制效应引起我们关注。本研究旨在探讨重组人血管内皮抑素（recombinant human endostatin injection）对非小细胞肺癌组织中血管内皮生长因子（vascular endothelial growth factor, VEGF）-C、VEGF-D和VEGF受体（VEGFR）-3表达及对外周血循环肿瘤细胞数目的影响。方法荷瘤裸鼠随机分为空白对照组、顺铂组、不同浓度重组人血管内皮抑素组及重组人血管内皮抑素+顺铂组，连续给药2周。1周后检测肿瘤组织中VEGF-C、VEGF-D、VEGFR-3的表达水平和微淋巴管密度。免疫荧光染色诊断和计数循环肿瘤细胞。结果重组人血管内皮抑素组与重组人血管内皮抑素联合顺铂组的表达阳性率、微淋巴管密度均明显低于空白对照组与顺铂组（P < 0.05）；较高浓度的重组人血管内皮抑素联合顺铂组与重组人血管内皮抑素组表达阳性率和微淋巴管密度低于相应较低重组人血管内皮抑素浓度的组别（P < 0.05）。各组微淋巴管密度与VEGF-C、VEGF-D、VEGFR-3表达阳性率存在正相关。重组人血管内皮抑素联合顺铂各组的循环肿瘤细胞数目明显低于单独使用顺铂或重组人血管内皮抑素（P < 0.05）。结论重组人血管内皮抑素可以抑制肿瘤新生淋巴管生成，减少循环肿瘤细胞，作用大小与浓度有关。与顺铂联合使用能够更有效的减少循环肿瘤细胞。

Endostar, a novel human recombinant endostatin, attenuates liver fibrosis in CCl4-induced mice.

Decreasing hepatic fibrosis remains one of the major therapeutic challenges in hepatology. The present study aims to evaluate the effect of Endostar on both CCl4-induced liver fibrosis in mice and a hepatic stellate cell (HSC) line. Two main models were studied: (i) a liver fibrosis model was induced in BALB/c mice using CCl4 by intraperitoneal injection for six weeks. Six animal groups were studied: group 1: normal animals; group 2: CCl4-induced liver fibrosis; group 3: CCl4 + Endostar 20 mg/kg/d, six weeks; group 4: CCl4 + Endostar 10 mg/kg/d, six weeks; group 5: CCl4 + Endostar 20 mg/kg/d, four weeks; group 6: CCl4 + Endostar 10 mg/kg/d, four weeks corresponded to different Endostar doses and duration of administration. Liver fibrosis was evaluated by histopathological staining and liver hydroxyproline content. Expressions of collagen type I, α-smooth muscle actin (α-SMA), TGF-β1 and VEGFR were measured by real-time polymerase chain reaction (PCR). (ii) A liver cell model. HSC-T6 cells were cultured with or without Endostar for 12 h or 24 h. Expressions of collagen type I, α-SMA, and TGF-β1 were measured by real-time PCR. Collagen I and transforming growth factor β1 (TGF-β1) contents in cell supernatant were measured by enzyme-linked immunosorbent assay. As compared to the group without Endostar, liver fibrosis scores and hydroxyproline content were decreased in both Endostar groups (P < 0.05). Moreover, Endostar inhibited the hepatic expression of α-SMA, TGF-β1, Collagen-1, VEGFR1, and VEGFR2 mRNA (P < 0.05). In the HSC-T6 cell line model, Endostar profoundly inhibited the expression of α-SMA, Collagen-1, and TGF-β1 mRNA. Expressions of Collagen-1 and TGF-β1 protein were decreased in the Endostar group as compared to the normal controls in the supernatant of HSC-T6 cells (P < 0.05). Endostar decreased both liver fibrosis in CCl4-induced mice and collagen synthesis in HSCs invitro. Therefore, this recombinant human endostatin is a promising compound for counteracting liver fibrosis.

The evaluation of anti-angiogenic effects of Endostar on rabbit VX2 portal vein tumor thrombus using perfusion MSCT.

BackgroundThere were many treatments for hepatocellular carcinoma with portal vein tumor thrombus (PVTT), in which targeted anti-angiogenic drug therapy is becoming a popular research topic. However, an objective and non-invasive method that can evaluate the treatment effects is still lacking.MethodsEighteen New Zealand white rabbits implanted with VX2 tumor thrombus in portal vein were randomly assigned into 3 groups: Endostar, saline, or control, six in each group. Multi-slice CT (MSCT) perfusion scanning was performed to measure the differences in blood flow (TBF), tissue blood volume (TBV), and capillary permeability time the surface (PS) before and after Endostar treatment, between Endostar and saline treatment. Two weeks after treatment, both Endostar and saline groups underwent CT perfusion scan. The rabbits then were sacrificed by air embolism, and specimens of tumor thrombosis were collected. Immunohistochemistry assay was also performed to compare the expression of vascular endothelial growth factor (VEGF) in PVTT after Endostar, saline and placebo treatment.ResultsIn Endostar group, PVTT CT perfusion parameters (TBF, TBV, PS) significantly decreased after the treatment (p <0.05). Post-treatment PVTT CT perfusion parameters (TBF, TBV, PS) were significantly lower in Endostar group than in Saline group (p <0.05). VEGF is mainly expressed in cytoplasma. After Endostar treatment, the expression of VEGF in PVTT was markedly reduced. There was also significant difference on post-treatment VEGF protein expression measured by Immunohistochemistry assay between Endostar group and control group (p <0.05). Post-treatment PVTT CT perfusion parameters (TBF, TBV, PS) were positively correlated with VEGF protein expression in all 3 groups (rs > 0, p <0.05).ConclusionsMulti-slice CT perfusion imaging can evaluate the anti-angiogenic effects of Endostar for the VX2 tumor thrombus in portal vein, and provide quantitative functional information.

Experimental Study of Endostar Injection Concomitant with Cryoablation on Lung Adenocarcinoma A549 Xenografts

To explore the inhibiting effect and mechanism of Endostar injection concomitant with cryoablation on lung adenocarcinoma A549 xenografts in nude mice. A total of 24 nude mice with subcutaneous xenografts of the A549 cell line were established and divided into 4 groups when the maximal diameters of tumors became 1 cm: control group, Endostar group, cryoablation group and combination group (Endostar concomitant with cryoablation). The nude mice were sacrificed after 21-days treatment, tumour tissues were removed to measure their volume, in situ test of TdT-mediated dUTP nick end labeling (TUNEL) was adopted to determine the cellular apoptosis around freezing injury zones, and immunohistochemical SP test was applied for the detection of micro-vessel density (MVD) and vascular endothelial growth factor (VEGF) expression levels. At 21-days after treatment, the growth velocities of control group, Endostar group, cryoablation group and combination group were 236.7 ± 51.2%, 220.0 ± 30.6%, 159.5 ± 29.3% and 103.3 ± 25.5% (P<0.01), while cellular apoptosis rates of tumors were 21.7 ± 2.34%, (22.17 ± 1.47)%, 38.3 ± 1.37% and 49.2 ± 1.72%, (P<0.01), respectively, according to the immunohistochemical test. MVD and VEGF expression levels in the combination group were both lower than in other groups (P<0.01), also being positively related (r=0.925, P<0.01). Endostar can significantly improve the inhibitory effects of cryoablation on xenografts of lung adenocarcinoma A549, and the mechanism is probably associated with its function as an inhibitor of tumour neo-angiogenesis through down-regulating VEGF expression.

Evaluating the effects of an angiogenesis inhibitor of endostar with a novel RGD dimer probe labeled by 99Tcm

Objective To study the feasibility of a novel probe 99Tcm-HYNIC-2(poly-(ethylene glycol),PEG) 4-Dimer (Dimer:E-[c (RGDfK) 2]) as a potential imaging agent for integrin αv β3 positive tumors,and also to observe the influence of an angiogenesis inhibitor,endostar,on the biodistribution and tumor uptake of the tracer in tumor bearing nude mice.Methods The expression of integrin αv β3 in human glioma cells U87MG was determined with immunofluorescence staining before and after treatment with endostar.99Tcm-HYNIC-2PEG4-Dimer was prepared and administered in U87MG tumor bearing mice in 6 h after either administration of endostar (200 μl) or saline (control group) and then biodistribution study was performed.Other 16 mice were divided into endostar treated group (20 mg/kg) and control group (saline) and then gamma imaging was performed in the two groups.Statistical significance of differences between the two groups was assessed using two-sample t test.Results Radiochemical purity of 99Tcm-HYNIC-2PEG4-Dimer was exceeded 95％.The expression of integrin αvβ3 in U87MG cell was high and gradually decreased after treatment with endostar.There was a negative dose-effect relationship between the dose of endostar and the expression of integrin αvβ3 with the peak effect at the dose of 400 μg/ml.The distribution study in vivo showed that the tracer uptake of U87MG tumors was high,but it decreased after injection of endostar.At 90 min,the ％ID/g of endostar and control groups were 1.50±0.08 and 6.27±0.33,respectively (t =40.23,P＜0.05).The average T/NT ratios of 99Tcm-HYNIC-2PEG4-Dimer uptake in the endostar and control groups were 1.02±0.11 and 2.58±0.36,respectively (t =10.25,P＜0.05).The integrin αv β3 positive expression ratios of tumor in endostar and control groups were (33.1 ±2.7) ％ and (81.5±3.2) ％,respectively (t =32.60,P＜0.05).Conclusions The novel probe 99Tcm-HYNIC-2PEG4-Dimer may be a promising radiotracer for integrin αvβ3-positive tumor imaging.It may be used for monitoring the therapeutic effect of endostar and may be potentially used for screening the candidates of anti-angiogenesis therapy. Key words: Glioma; Arg-Gly-Asp; Radionuclide imaging; Angiogenesis inhibitors ; Mice, nude

Effect of endostar combined with cisplatin on expression of VEGF and Sema3A of Lewis lung cancer rats

ObjectiveTo evaluate the therapeutic effect of endostar (ED) combined with cisplatin(DDP) on model of C57BL/6 rats, and to further investigate the inhibiting mechanism of endostar from tumor angiogenesis. MethodsLewis lung cancer cells were inoculated in C57BL/6 mouse, then the mouse were randomized into control group (group A), ED (group B), DDP (group C) and ED/DDP (group D). They were treated according to the plan. And the expressions of VEGF and Sema3A were evaluated by immunhistochemisty. ResultsThe weight of tumor increased in group A and B. It was decreased in group C and D. The tumor volume was increased in all the 4 groups. The VEGF expression of group D was obviously lower than the other group 3, but the Sema3A expressed of group D was significantly strengthener than the other group 3. The VEGF expression of group B and group D were obviously low especially in the 4th-8th days. Pearson correlated analysis showed that the expression VEGF and Sema3A were negatively correlated (r=-0.72, P<0.05). ConclusionsED combined with DDP could control the tumor growth effectively, and avoid weight loss. ED could reduce VEGF expression, and enhance Sema3A expression. Tumor vessel presents transient normalization. It is easy for DDP perfusion, and to kill tumor cells.

The inhibitory effects of Endostar combined with chemotherapy on human esophageal squamous cell carcinoma xenograft in mice

The purpose of this study was to investigate the efficacy of Endostar combined with chemotherapy on human esophageal squamous cell carcinoma Eca-109 in mice. The tumor xenograft models were established and randomly assigned to 4 groups: control group, Endostar group (1.5 mg/kg day, once daily for 3 weeks), chemotherapy group (Paclitaxel 10 mg/kg day, Cisplatin 5 mg/kg day, for 1, 7, 14, 21 days), and chemotherapy + Endostar group (combination). The length and width of tumor were measured and the tumor volumes (cm(3)) were calculated every other day. Three weeks later, the mice were executed, and the tumor tissues were collected to weigh and analyse the histopathology and proliferation for tumor xenograft. The results demonstrated that the tumor volumes and weight in chemotherapy + Endostar group were significantly lower than that in other three groups. Meanwhile, the cell proliferation of tumor xenograft in combined treatment group was significantly lower than that in other three groups. It was concluded that Endostar combined with chemotherapy could obviously enhance the inhibitory effect on esophageal squamous cell carcinoma Eca-109 in mice.

1337 - NP Chemotherapy Plus Endostar Compared with NP Alone as First-Line Therapy in Stage IIIB/IV Non-Small Cell Lung Cancer: A Retrospective Study

ABSTRACT Purpose Recombinant human endostatin is a novel inhibitor of tumor angiogenesis. Previous studies have indicated that Recombinant human endostatin (endostar) plus vinorelbine-cisplatin chemotherapy could improve objective response rates (ORR) and time to progression (TTP) of advanced non-small cell lung cancer (NSCLC) patients with decreased toxicity. The purpose of this study was to retrospectively compare the efficacy and safety of NP chemotherapy plus endostar with NP alone as first-line therapy in stage IIIB/IV non-small cell lung cancer. Methods We reviewed the records of 65 previously untreated Chinese patients with stage IIIB/IV NSCLC who were treated with NP chemotherapy plus endostar or NP alone between January 2005 and December 2010 at The First Hospital of China Medical University. Vinorelbine (25mg/m2) was administered on days 1 and 8, cisplatin(75mg/m2) was administered on day 1, and endostar (7.5mg/m2) was administered on days 1-14. Treatments were repeated every 3 weeks for a maximum of 6 cycles. The best tumor response was evaluated according to Response Evaluation Criteria in Solid Tumors (RECIST 1.1). The adverse events were defined by NCI-CTC 3.0 criteria. Univariate and multivariate analyses were performed using the SPSS 20.0 software. Results The median age of patients was 58 years (range, 34–77 years). Our main results are as follows: Addition of endostar to standard NP chemotherapy significantly reduced the risks of disease progression and death. The median progression-free survival of the NP plus endostar group and NP group were 7.4 months and 5.5 months (P= 0.042). The median overall survival of the NP plus endostar group and NP group were 17.6 months and 12.6 months (P = 0.046). There was no statistical difference in objective response rates and disease control rates between the two groups. The incidence of grade 3/4 hematologic adverse events was higher in NP plus endostar group (48.5% vs 21.9%, P = 0.025) but tolerable. Conclusion This study demonstrated that the addition of endostar to the NP regimen is an effective treatment option with an acceptable increase in hematologic toxicity in previously untreated patients with stage IIIB/IV non-small cell lung cancer. Disclosure All authors have declared no conflicts of interest.

Randomized phase II study of recombinant human endostatin in combination with chemotherapy in previously untreated extensive-stage small-cell lung cancer (NCT00912392).

7091 Background: Endostar (recombinant human endostatin) is a novel antiangiogenesis drug developed in China for non-small cell lung cancer (NSCLC). Because of promising efficacy signals, we performed a randomized phase II trial to assess the efficacy and safety of adding endostar to first-line standard chemotherapy for treatment of chemonaive extensive-stage small-cell lung cancer (SCLC). Methods: Extensive-stage SCLC patients with a performance status 0-2 were randomly assigned to endostar group (endostar 7.5mg/m2 D1-D14 with carboplatin AUC=5 plus etoposide 60mg/m2 D1-D5 of a 21-day cycle for six cycles) or the control group (the same dose of carboplatin plus etoposide). Patients in endostar treatment group with CR, PR and SD were treated with single-agent endostar until progression or unacceptable toxicity. The primary end point is progression-free survival (PFS). The secondary end points are overall survival (OS) and response rate (RR). Results: 140 patients were enrolled from June 2009 to June 2011, and 137 patients were included in full analysis set. 68 patients were randomly assigned to the endostar treatment and 69 patients to the control group. Median age was 57 years and 80.9% for male in the endostar group while median age was 58 years and 85.5% for male in the control group. Median PFS was similar for endostar and control group (6.2 v 5.9 months, P=0.163, HR 0.762; 95%CI 0.519-1.119). Median overall survival (OS) was also similar for both groups (12.4 v 12.3 months, P=0.475, HR 0.835; 95%CI 0.508-1.371). Overall RR were 76.5% for endostar group and 68.1% for the control group (p=0.275). 20 patients in the endostar group completed six cycles of therapy and subsequently treated with single-agent endostar as maintenance therapy and the median PFS and OS were 6.8 and 12.4 months respectively. The rate of ≥ 3 grade adverse events occurred in both groups was similar and no new or unexpected safety signals for endostar were observed. Conclusions: The addition of endostar to carboplatin plus etoposide for treatment of extensive stage SCLC didn’t improve the PFS significantly, with an acceptable toxicity profile. And no improvement in OS was observed.

Protection against acute radiation-induced lung injury: A novel role for the anti-angiogenic agent Endostar

Radiotherapy is commonly used to treat thoracic malignancies, but often causes severe lung injury. Currently there are no effective protective strategies against radiation-induced lung injury (RILI). This study aimed to evaluate the ability of an angiogenesis antagonist, Endostar, against RILI, and the underlying mechanism in a mouse model. A total of 108 C57BL/6 female mice were randomized into 4 groups (n=27): i) control group; ii) Endostar group, animals were administered 7.75 ml/kg Endostar through intraperitoneal injection; iii) irradiation group, RILI was induced by exposing the animals to a single external irradiation on the thoraces (6 MV X-ray, 12 Gy); and iv) irradiation plus Endostar group, animals were subjected to Endostar treatment and irradiation as in groups 2 and 3. A total of 3 animals from each of the 4 groups were sacrificed at 1, 6, 12, 24 and 72 h and at 2, 4, 8 and 24 weeks following treatment. Clinical signs and pathology of RILI were examined. The expression of transforming growth factor-β 1 (TGF-β1) in lungs was analyzed by real-time quantitative polymerase chain reaction (RT-QPCR) and immunohistochemistry. Compared with the control group, irradiation induced evident interstitial edema and a significant increase in inflammatory cells in the lungs (P<0.05). Correlated with these changes, a notable increase in TGF-β1 mRNA level and a robust increase in TGF-β1 immunoreactivity were observed in lung tissues in a time-dependent manner following irradiation (P<0.05). Endostar administration effectively attenuated the magnitude of the increase in inflammatory cells as well as the elevation of TGF-β1 expression in lung tissues after RILI (P<0.05). In conclusion, radiation induced an increased expression of the inflammatory mediator TGF-β1 and the associated pathogenesis in the lung, while Endostar was able to at least partially attenuate RILI through downregulating the expression of TGF-β1 in mice. Our findings suggest that Endostar may be a novel protective agent against RILI.

重组人血管内皮抑素联合放疗对肺腺癌A549细胞TGF-β1、HIF-1α表达的影响

通过观察Endostar联合放疗对肺腺癌A549细胞增值及TGF-β1、HIF-1α表达的影响，初步探讨重组血管内皮抑素的放疗增敏机制。方法：将培养至对数生长期的A549细胞分为对照组、单纯放疗组、恩度组、联合组，检测各组A549细胞增殖率及TGF-β1、HIF-1α蛋白和mRNA表达情况。结果：1) 除对照组外，各组细胞的增殖随时间的改变都受到不同程度的抑制，以联合组最为明显，差异具有统计学意义(p ；2) 处理后各组A549细胞TGF-β1、HIF-1α的分泌下降，24 h以放疗组及联合组TGF-β1、HIF-1α表达量明显下降 (p ；3) PCR扩增后，各处理组 TGF-β1、HIF-1α mRNA的表达随着时间变化均表现出下降趋势，联合组在24 h差异有显著统计学意义(p 。结论：恩度联合放疗可显著抑制肺腺癌A549细胞增殖及TGF-β1、HIF-1α的表达，恩度可能通过下调TGF-β1、HIF-1α的表达进而发挥放疗增敏作用。 To observe the effect of Endostar combined with Radiotherapy on the expression of Transforming Growth Factor-beta 1 (TGF-β1) and Hypoxia-induced factor-1 (HIF-1α) in lung adenocarcinoma A549 cells and to discuss the possible mechanisms. Methods: To culture the A549 cell, when the cells achieve Logarithmic phase and there are four groups: Control group; Endostar group; Radiotherapy group; Endostar in combination with Radiotherapy. Cell proliferate rate was measured by CCK8, and the expression level of TGF-beta 1 and HIF-1α protein and mRNA of A549 cells in each group were examined by ELISA and PCR. Result: 1) Besides control group, the proliferation of tumor cells in each group are subject to different degree of inhibition. Compared with control and Endostar groups, there is a significant difference of proliferate inhibition in Endostar combined with Radiotherapy group (p β1 and HIF-1 of A549 cells in each group appears to be declining differently. The Radiotherapy group and combined group have a significant difference in cell proliferation at 24 hours, compared to control group (p β1 and HIF-1α mRNA over time (0 - 24 hours) compared with the control group, all treatment groups showed a decreasing trend. After 24 hours of treatment, Compared with control group and Endostar group, the difference in the combined group was statistically sig- nificant (p Conclusion: Endostar combined with Radiotherapy could significantly enhance the inhibition of proliferate of cells and expressions of TGF-β1 and HIF-1α in tumor cells. Thus enhance the inhibition of tumor angio- genesis and metastatic.

Combination Therapy of Transcatheter Arterial Chemoembolization and Arterial Administration of Antiangiogenesis on VX2 Liver Tumor

This study was designed to evaluate the antitumorigenic efficiency of Endostar (an antiangiogenic agent) arterially administrated combined with transcatheter arterial chemoembolization (TACE) on liver tumor, and validation of perfusion CT for quantitative measurements of the results. Thirty rabbits bearing VX2 liver tumors were randomly and equally distributed into three groups. One of the following treatment protocols was performed in each group: 1) group 1 was treated with TACE and simultaneously arterially administrated Endostar; 2) group 2 with TACE alone, and 3) a control group that had saline injected through hepatic artery. Routine CT scan was performed before treatment, and perfusion CT imaging was performed 2 weeks after treatment. Immunohistochemical biomarkers of microvascular density (MVD) and the expression of vascular endothelial growth factor (VEGF) were measured for assessments of angiogenesis. We observed a statistically significant reduction from the control in the volume, growth rate, and size of the tumor 2 weeks after treatment with both TACE plus Endostar and with TACE alone (P < 0.01). Although there was no statistically significant difference in tumor size between the group with TACE plus Endostar and the group with TACE alone (P > 0.05), MVD and VEGF were significantly less expressed in the TACE plus Endostar group than both groups with TACE alone and the control group (P < 0.01). Blood flow (BF), blood volume (BV), and permeability-surface area products (PS) in the group with TACE plus Endostar on perfusion CT were significantly higher than other two groups (P < 0.05), which were positively correlated with the MVD and VEGF values (P < 0.05). TACE with arterial administration of Endostar simultaneously significantly inhibited the angiogenesis biomarkers associated with TACE in a rabbit model bearing VX2 liver tumor, which indicates that the combined treatment protocol may have potential synergistic effects on liver cancer. It also is suggested that perfusion CT may be useful for monitoring antiangiogenic/antivascular treatment in the liver tumors.

Effect of photodynamic therapy and endostatin on human glioma xenografts in nude mice

The aim of this study was to investigate the effect of endostar (a recombinant human endostatin) and photodynamic therapy (PDT) on gliomas. To establish glioma xenografts, human U251 glioma cells were injected into the brain of nude mice. Mice with MRI-confirmed glioma received hematoporphyrin monomethyl ether (HMME)-mediated PDT, daily injection of endostar or their combination, respectively. After treatment, tumor volume, the expression of HIF-1α, VEGF-A and apoptosis marker, and animal survival were examined. PDT and endostar treatment can prolong survival. Changes in induction of apoptosis, tumor growth and survival were more significant in PDT+endostar group. After PDT, HIF-1α and VEGF-A expressions were markedly increased. After endostar treatment, HIF-1α and VEGF-A expressions were significantly reduced. PDT in combination with endostar can significantly inhibit the growth of glioma xenografts. This approach may represent a promising strategy in the treatment of glioma.

Recombinant human endostatin inhibited the lymphangiogenesis and lymph node metastasis in a mouse xenograft model of colon cancer

Objective To observe the effects of recombinant human endostatin, Endostar, on tumor lymphangiogenesis and lymph node metastasis in a mouse xenograft model of colon cancer. Methods Human colon cancer SW620 cells were implanted subcutaneously into the hind flank of the male nu/nu mice to estab-lish a xenograft model. The control group was treated with normal saline, and the Endostar group was treated with Endostar by subcutaneous injection every other day. Results (1) The rate of lymph node metastasis was 10% (1/10) in the Endostar group, and 80% (8/10) in the control group (P <0.01 ) ; (2) The den-sity of lymphatic vessels in the Endostar group was decreased significantly as compared with the control group (7.17 ± 0. 75 vs. 14.83 ± 0.98 per field, P < 0.05 ) ; (3) Endostar inhibited the expression of vascular endo-thehal cell growth factor (VEGF)-C and VEGF-D proteins in tumor tissues and SW620 cells. Conclusion Recombinant human endostatin suppressed lymphangiogenesis and lymph node metastasis by inhibiting the ex-pression of VEGF-C and VEGF-D in a mouse xenograft model of colon cancer. Key words: Endostatin; Colorectal carcinoma; Lymphatic vessel; Metastasis

The inhibitory effects of rh-endostatin (YH-16) in combination with radiotherapy on lung adenocarcinoma A549 in mice and the underlying mechanisms

In order to investigate the inhibitory effects of Endostar (rh-endostatin, YH-16) in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy, the transplantation tumor models of A549 lung adenocarcinoma were established. When the largest diameter of tumor reached 1.0 cm, all nude mice were randomly divided into 4 groups: Endostar group, radiotherapy group, radiotherapy plus Endostar (combined treatment) group, and control group (n=6 in each group). The largest diameter and the vertical diameter of tumor were measured at different time points. At the 16th day, mice were executed, and the tumors were applied to analysis of rate of tumor cell apoptosis, and the expression levels of basic fibroblast growth factor (bFGF) mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and those of vascular endothelial growth factor (VEGF) by immunohistochemistry. The results demonstrated that the rate of tumor inhibition in combined treatment group was higher than that in other groups. And the rate of tumor cell apoptosis in combined treatment group was also higher than that in other groups. Meanwhile, the levels of bFGF mRNA and VEGF expression in combined treatment group were lower than those in other groups. It was concluded that Endostar obviously enhanced the curative effectiveness of radiotherapy on lung adenocarcinoma A549 in mice. The underlying mechanisms may involve the down-regulation of bFGF mRNA and VEGF expression to inhibit angiogenesis by Endostar and the cooperative effect of Endostar and radiotherapy to synergistically promote tumor cell apoptosis. And Endostar inhibits angiogenesis by down-regulating the expression of bFGF mRNA and VEGF.

Changes of activated circulating endothelial cells and survivin in patients with non-small cell lung cancer after antiangiogenesis therapy

Although antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells (aCECs) and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy. Patients of non-small cell lung cancer (NSCLC) treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction (PCR). After treatment, the amount of activated CECs decreased significantly in clinical benefit cases (P = 0.021 in chemotherapy alone, P = 0.001 in chemotherapy plus Endostar), increased in disease progressive cases (P = 0.015 in chemotherapy alone, but P = 0.293 in chemotherapy with Endotatar). After therapy, the expression of survivin mRNA decreased in clinical benefit cases (P = 0.001) and increased in disease progressive cases (P = 0.018). A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy (P = 0.001 and 0.021, respectively), but only in the chemotherapy with Endostar group pre-therapy (P = 0.030) rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression (TTP) (r = 0.322, P = 0.012); a negative correlation was found between the amount of survivin mRNA in serum post-therapy and TTP (r = -0.291, P = 0.048). Activated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.