Effect Of Recombinant Human Endostatin Research Articles

Alleviation of Papain-Induced Osteoarthritis by Recombinant Human Endostatin via Downregulation of Matrix Metalloproteinase-13, Interleukin-1 and Interleukin-6 in Rats

Osteoarthritis (OA) is a degenerative disease of joints commonly occurring in the elderly and middleaged people. This study aimed to investigate the effect of recombinant human endostatin (rhEndo) on OA and the levels of MMP-13, IL-1 and IL-6 in the synovial fluid in osteoarthritis rats. OA models were made by injecting 4% papain into the knee joint cavity of rats once every three days for three times. The models were then injected subcutaneously with rhEndo and examined six weeks later for the Mankin scores and levels of MMP-13, IL-1 and IL-6 using ELISA. Compared with control, the Mankin score as well as the levels of IL-1, IL-6 and MMP-13 were significantly increased in the models (0.30 vs. 5.80, 1.12 vs. 12.84 pg/ mL, 12.22 vs. 43.82 pg/ mL and 0.23 vs. 26.31 ng/ mL). Following treatment with 4 mg/kg rhEndo, the Mankin score in model decreased to 0.90, meanwhile, the levels of IL-1, IL-6 and MMP-13 decreased significantly to 0.79 pg/ mL, 2.89 pg/mL and 1.17 ng/mL, respectively, in a dose dependent manner. Therefore, rhEndo can alleviate osteoarthritis by reducing MMP-13, IL-1 and IL-6 expression in rats.

Endostar improved efficacy of concurrent chemoradiotherapy with vinorelbine plus carboplatin in locally advanced lung squamous cell carcinoma patients with high serum Lp(a) concentration.

The role of vascular targeting therapy combined with concurrent chemoradiotherapy (CRT) has produced many inconsistent results in locally advanced non-small-cell lung cancer (NSCLC), especially in lung squamous cell carcinoma (LSCC). Lipoprotein (a) [Lp(a)] may be critical in the development of tumor angiogenesis, and its levels are individualized and determined genetically. This study aimed to determine whether Lp(a) is correlated with effects of recombinant human endostatin (Endostar) combined with concurrent CRT for locally advanced LSCC. Patients with locally advanced LSCC from December 2008 to December 2017 were retrospectively analyzed. Patients were divided into two groups: (I) a chemoradiotherapy group (CRT group) which received weekly vinorelbine and carboplatin concurrently with radiotherapy 60Gy, and (II) an Endostar in combination with chemoradiotherapy group (ECRT group) which received Endostar intravenous drip for 1-14 days (every 3 weeks) concurrently with CRT. Fasting venous blood samples for serum Lp(a) in all patients were collected before the treatment. The effect of Endostar was assessed by stratified analysis. A total of 94 patients were recruited in this study. There were 59 cases in the CRT group and 35 cases in the ECRT group. Overall, the median progression-free survival (PFS) was 9.6 vs. 14.2 months (P=0.0671), and the overall survival (OS) was 15.0 vs. 20.6 months (P=0.114), in the CRT and ECRT groups respectively. The median of Lp(a) was 218 mg/L. In patients with serum Lp(a) less than 218 mg/L, the median PFS was 10.0 vs. 9.4 months (P=0.406), and the OS was 15.4 vs. 16.3 months (P=0.958) in the CRT and ECRT groups, respectively. However, in patients with serum Lp(a) higher than 218mg/L, the median PFS was 9.0 vs.15.8 months (P=0.011), and the OS was 14.0 vs. 21.1 months (P=0.055), in the CRT and ECRT groups, respectively. Cox proportional hazard model analysis revealed that a high concentration of Lp(a), ≥218 mg/L, is a prognostic factor for PFS [hazard rate (HR), 0.43 (0.23-0.81)] and OS [HR, 0.52 (0.27-0.98)] in locally advanced LSCC (P<0.05). The serum concentration of Lp(a) may serve as a biomarker to identify the patients who would benefit from Endostar treatment with concurrent CRT in stage III LSCC.

Clinical observation of recombinant human endostatin combined with cisplatin in the treatment of pleural effusion of lung cancer

Objective To investigate the clinical effect of recombinant human endostatin combined with cisplatin in the treatment of malignant pleural effusion of lung cancer. Methods 100 lung cancer patients with malignant pleural effusion were selected and divided into observation group and control group according to the random number table method, with 50 cases in each group.The control group was treated with cisplatin 40mg/m2 single-dose pleural perfusion.The observation group was treated with 40mg/m2 of cisplatin on the basis of 45mg of recombinant human endostatin.All the patients were treated twice a week, treatment for 3 weeks.The clinical efficacy, adverse reactions, the immune function indicators before and after treatment, quality of life improvement were compared between the two groups. Results (1)In the control group, 10 cases were cured, markedly effective in 24 cases, effective in 12 cases, the total effective rate was 92%.In the observation group, 4 cases were cured, markedly effective in 14 cases, effective in 18 cases, the total effective rate was 72%, the difference between the two groups was statistically significant (χ2=9.15, P 0.05), but the number of adverse reactions in the observation group was less than that in the control group, which indicated that the safety of the observation group was high.(3) Before treatment, the CD4+, CD8+ and CD4+/CD8+ between the two groups had no statistically significant differences (t=0.03, 0.26, 0.67, all P>0.05). After treatment, the percentage of CD4+ and CD4+/CD8+ in the observation group were higher than those in the control group, the percentage of CD8+in the observation group was lower than that in the control group, the differences were statistically significant (t=11.58, 12.36, 10.84, all P<0.05). (4) In the observation group, improved in 31 cases, stable in 10 cases, decreased in 9 cases.In the control group, improved in 25 cases, stable in 12 cases, decreased in 13 cases.There was in statistically significant difference between the two groups (χ2=2.30, P<0.05). Conclusion Recombinant human endostatin combined with cisplatin in the treatment of pleural effusion of lung cancer is significant, which can enhance the immune function of patients, improve the quality of life of patients, and with good safety, less adverse reactions. Key words: Recombinant human vascular endothelin; Cisplatin; lung cancer; Pleural effusion; Clinical efficacy

Recombinant human endostatin combined with radiotherapy inhibits colorectal cancer growth

BackgroundTo examine the effects of recombinant human endostatin combined with radiotherapy on colorectal cancer HCT-116 cell xenografts in nude mice.MethodsForty male BALB/c nude mice were injected with human colorectal cancer HCT-116 cells to form xenografts and then randomized into the following 4 groups (each group comprised ten mice): a control group, an endostatin group (20 mg/kg endostatin once a day for 10 days), a radiotherapy group (a 6-Gy dose was administered via a 6-MV X-ray on day 5 post-inoculation), and a combination therapy group (radiotherapy with endostatin treatment). The tumor growth inhibition rate were detected. CD31, vascular endothelial growth factor (VEGF), and hypoxia inducible factor-1α (HIF-1α) expression and microvascular density (MVD) were evaluated by immunohistochemistry. The expression of VEGF protein was also detected by western blotting.ResultsThe tumor growth inhibition rate in the radiotherapy with endostatin treatment group was significantly higher than those in endostatin group or radiotherapy group (77.67% vs 12.31% and 38.59%; n = 8 per group, P < 0.05). The results of immunohistochemistry showed that treatment with radiotherapy induced significant increases in CD31, VEGF, and HIF-1α expression and MVD compared with treatment with saline, while treatment with endostatin or radiotherapy with endostatin induced reductions in CD31, VEGF, and HIF-1α expression and MVD compared with treatment with saline (n = 8 per group, P < 0.05). The results of western blotting showed that VEGF protein expression in radiotherapy group was significantly increased compared with that in the control group. However, VEGF protein expression in the endostatin or radiotherapy with endostatin groups was significantly decreased compared with that in the control group (n = 8 per group, P < 0.05).ConclusionsEndostatin combined with radiotherapy can significantly inhibit HCT-116 cell xenograft growth, possibly by inhibiting angiogenesis and attenuating tumor cell hypoxia.

Antiangiogenic effects of recombinant human endostatin in lung cancers.

Antiangiogenic therapy, as a new anticancer method, can improve the anticancer effect of traditional therapies. Different antiangiogenic drugs may have different vascular normalization time windows. Whether the antiangiogenic treatment is within the vascular normalization time window is very important in the treatment of cancers. Previous studies have indicated that recombinant human endostatin (rh-ES) can transiently normalize tumor microvessels. Yet the molecular mechanism and the time window of rh-ES remains unclear. The aim of the present study was to explore the optimal time window and molecular mechanism of rh-ES in inhibiting Lewis lung cancer (LLC). By comparatively accessing the changes in microvascular and hypoxic conditions of tumors in host mice treated with rh-ES or saline for different days, the authors aimed to investigate the best administration time of rh-ES treatment on human lung cancers and obtain a better understanding concerning the involved molecular mechanism. A total of 40 C57/BL6 mice with LLC xenografts were randomly divided into normal saline (NS) and rh-ES groups (20 mice/group). 0.2 ml NS or 5 mg/kg rh-ES were administrated via intraperitoneal injection (i.p.) into each mouse each day during the 9-day experiment. A total of 5 mice from each group were sacrificed at day 2, 4, 6 or 9. CA9 and RGS5 expression levels of both groups were compared using immunohistochemistry, reverse transcription-quantitative polymerase chain reaction and ELISA. Rh-ES caused vascular normalization and improved hypoxia at days 4 and 6. Compared with the control (NS) group, both CA9 and RGS5 expression in rh-ES group were significantly decreased at days 4 and 6 (P<0.05), while no significant change between two groups was observed at days 2 and 9. Rh-ES can induce transient tumor vascular normalization and improves tissue hypoxia in LLC tumors. The vascular normalization window is accompanied by the reduction in RGS5 and CA expression.

Effect of recombinant human endostatin on hypertrophic scar fibroblast apoptosis in a rabbit ear model

Hypertrophic scar (HS) is a dermal fibroproliferative disorder characterized by the excessive proliferation of fibroblasts and is thought to result from a cellular imbalance caused by the increased growth and reduced apoptosis of hypertrophic scar fibroblasts (HSFs). Our recent study demonstrated that recombinant human endostatin (rhEndostatin) plays a key role in the inhibition of HSF proliferation in vitro, with a resulting decrease in dermal thickness and scar hypertrophy. However, the effect of this protein on HSF apoptosis is unknown. The present study was undertaken to directly examine the effect of rhEndostatin on HSF apoptosis in the rabbit ear model. Transmission electron microscopy and flow cytometry were used to investigate HSF apoptosis in scar tissues and cultured HSFs in vitro, respectively. The expression levels of the c-jun, c-fos, NF-κB, fas, caspase-3, and bcl-2 gene products in HSFs were quantified using real-time PCR and Western blotting assays. Our data reveal that rhEndostatin (2.5 or 5mg/ml) induces HSF apoptotic cell death in scar tissue. Additionally, HSFs treated with rhEndostatin (100mg/L) in vitro accumulated in early and late apoptosis and displayed significantly decreased expression of c-jun, c-fos, NF-κB, fas, caspase-3 and bcl-2. In sum, these results demonstrate that rhEndostatin induces HSF apoptosis, and this phenotypeis partially due to downregulation of NF-κB and bcl-2. These findings suggest that rhEndostatin may have an inhibitory effect on scar hypertrophy in vivo via HSF apoptotic induction and therefore has potential therapeutic use for the treatment of HS.

The accumulation of macrophages attenuates the effect of recombinant human endostatin on lung cancer.

BackgroundAlthough anti-angiogenic therapy is widely applied clinically, its efficacy has been less than expected. Screening for regulatory factors and sensitive indicators to define the effectiveness of these drugs is required. Through a retrospective study of clinical data, we found that patients with a higher peripheral monocyte-to-lymphocyte ratio (MLR) obtained less benefit from recombinant human endostatin (rhES, Endostar®), an anti-angiogenic drug, in lung cancer. Because MLR is positively correlated with macrophage count in tumors, this result suggests that macrophages may influence the effectiveness of rhES therapy in lung cancer.MethodsClinical data from 72 lung cancer patients treated with rhES were collected. Animal study, flow cytometry, immunofluorescence, enzyme-linked immunosorbent assay, Western blot analysis, and transwell migration assays were carried on Lewis lung carcinoma (LLC) cells, bone marrow-derived macrophages, macrophage cell line RAW264.7, and ANA-1 cells.ResultsClinical data showed that compared with the baseline MLR before rhES treatment, patients with progressive disease had higher MLRs than those of patients with partial response. Experimental results showed that more macrophages were recruited in the LLC tumors after rhES treatment and the majority of them displayed an M2-like phenotype. rhES aggravated hypoxia and the inflammatory response in the tumor microenvironment. Hypoxia promoted the expression of CCL2 by endothelial and fibroblast cells, which could induce macrophages recruitment, and increased levels of inflammatory cytokines (interleukin-4 [IL-4], IL-6, and IL-10) skewed macrophage polarization toward the M2-like phenotype. Hypoxia or inflammation cytokine-treated macrophages enhanced the progression of LLC in vitro and in vivo.ConclusionWe found rhES could aggravate hypoxia and the inflammatory response in the tumor microenvironment. These changes were favorable for macrophage accumulation, and skewed their polarization toward the M2-like phenotype which could help LLC to escape from the anti-angiogenic therapy. Thus, these data indicate the accumulation of macrophages in the tumor microenvironment may adversely affect the efficacy of rhES on lung cancer.

Inhibitory effect of recombinant human endostatin on the proliferation of hypertrophic scar fibroblasts in a rabbit ear model

Hypertrophic scar (HS) is a pathological scar that particularly occurs after traumatic injuries, surgical procedures and burning. Abnormal activation of hypertrophic scar fibroblasts (HSFs) intensifies fibrosis during wound healing. Our previous studies demonstrated that recombinant human endostatin (rhEndostatin) prevented synovial thickening in adjuvant arthritis (AA) rats via inhibition of proliferation and enhancement of apoptosis in synovial fibroblasts. However, the effect of this protein on HSF proliferation is not known. This study investigated the inhibitory effect of rhEndostatin on the proliferation of cultured HSFs in a rabbit ear model. MTT assay and flow cytometric detection were performed to investigate HSF proliferation and cell cycle progression, respectively. The expression levels of p53, p21, cyclinD1, cyclin-dependent kinase 4 (CDK4) and proliferating cell nuclear antigen (PCNA) in HSFs were detected using real-time PCR and Western blotting. Our data revealed that HSFs treated with rhEndostatin were significantly inhibited in a concentration-dependent manner with an IC50 value of 100mg/L. Also, rhEndostatin (100mg/L) primarily induced G0/G1 and partially G2/M cell cycle arrest of HSFs. There were significant decreases in the expression levels of p53, p27, CDK4, cyclinD1 and PCNA in HSFs treated with rhEndostatin. In conclusion, rhEndostatin inhibited HSF proliferation via G0/G1 and/or G2/M phase arrest of the cell cycle, which was partially due to the down-regulation of cyclinD1, CDK4 and PCNA. These findings suggest that rhEndostatin may reduce scar hypertrophy in vivo via inhibition of HSF proliferation and may be a novel agent for HS treatment.

Expression of Inhibitor of Differentiation-1 and Its Effects on Angiogenesis in Gastric Cancer.

To explore the effects of recombinant human endostatin (endostar, ES) and cisplatin on the growth of gastric cancer-transplanted tumor in nude mice and the expression of microvessel density (MVD). Human gastric cancer SGC-7901 cells were subcutaneously injected into the armpit of nude mice to prepare cancer-bearing nude mice. A total of 32 cancer-bearing nude mice were divided into four groups (each group with 8 mice). The four groups included control group and other three groups in which mice were treated with 5 mg/kg of ES (group E), 5 mg/kg of cisplatin (group Ci), and 5 mg/kg of ES combined with 5 mg/kg of cisplatin (group C), respectively. MVD was determined by immunohistochemistry, and the expressions of mRNA and protein of inhibitor of differentiation-1 (ID1) and vascular endothelial growth factor (VEGF) were detected with reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively. Apoptosis was observed with transmission electron microscope. Compared with control group, the sizes and weights of tumors were significantly decreased in other three groups (all p < 0.05). MVD was significantly lower in groups E, Ci, and C than in control group, and in groups E and C than in group Ci (all p < 0.05). Compared with control group, the expressions of mRNA and protein of ID1 and VEGF significantly decreased in groups E and C (all p < 0.05). There were no significant differences in the expressions of mRNA and protein of ID1 and VEGF between group Ci and control group. There was apoptosis in groups E and C, but no apoptosis was found in group Ci and control group. ES can inhibit the growth of gastric cancer cells through suppressing angiogenesis and promoting apoptosis of tumor cell. This study provides a new idea for the treatment of gastric cancer.

Recombinant human endostatin enhances the radioresponse in esophageal squamous cell carcinoma by normalizing tumor vasculature and reducing hypoxia.

The aim of this study was to investigate the effect of recombinant human endostatin (rh-Endo) in combination with radiation therapy (RT) on esophageal squamous cell carcinoma (ESCC) and explore the potential mechanisms. ECA109-bearing nude mice were administered RT and/or rh-Endo treatment. Tumor volume, survival, hypoxia and vascular parameters were recorded during the treatment schedule and follow-up as measures of treatment response. ESCC cell lines (ECA109 and TE13) and human umbilical vein endothelial cells (HUVECs) were developed to investigate the outcomes and toxicities of rh-Endo and RT in vitro. Hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) were also evaluated. In vivo studies of ECA109-bearing xenografts showed that rh-Endo improved the radioresponse, with normalization of tumor vasculature and a reduction in hypoxia. In vitro studies showed that rh-Endo did not radiosensitize ESCC cell lines but did affect endothelial cells with a time- and dose-dependent manner. Studies of the molecular mechanism indicated that the improved radioresponse might be due to crosstalk between cancer cells and endothelial cells involving HIF and VEGF expression. Our data suggest that rh-Endo may be a potential anti-angiogenic agent in ESCC especially when combined with RT. The improved radioresponse arises from normalization of tumor vasculature and a reduction in hypoxia.

Antitumor effect of recombinant human endostatin combined with cisplatin on rats with transplanted Lewis lung cancer.

To observe the antitumor effect and mechanism of recombinant human endostatin (Endostar) injection in tumor combined with intraperitoneal injection of cisplatin on subcutaneous transplanted Lewis lung cancer in rats. A total of 30 C57 rats were selected, and the monoplast suspension of Lewis lung cancer was injected into the left axilla to prepare the subcutaneous transplanted tumor models in the axilla of right upper limb. The models were randomly divided into Groups A, B, and C. Medication was conducted when the tumor grew to 400mm(3). Group A was the control group without any interventional treatment. Group B was injected with Endostar 5mgkg(-1)d(-1) for 10d. Group C was given the injection of Endostar 5mgkg(-1)d(-1) combined with intraperitoneal injection of cisplatin 5mgkg(-1)d(-1) for 10d. All the rats in three groups were executed the day after the 10 d medication and the tumor was taken off for measurement of volume and mass changes and calculation of antitumor rate, after which the vascular endothelial growth factor (VEGF) concentration in rats' plasma was determined by ELISA. The tumor tissues were cut for the preparation of conventional biopsies. After hematoxylin-eosin staining, the pathologic histology was examined to observe the structures of tumor tissues, VEGF score and microvessel density (MVD) in each group. The volume and mass of tumor in Groups B and C were significantly lower than Group A (P<0.05) while the tumor volume and mass in Group C were significantly lower than Group B (P<0.05). The antitumor rate in Group C was significantly higher than Group B (P<0.05), but the tumor VEGF score, MVD and plasma VEGF level in Group C were significantly lower than Groups A and B (P<0.05). In Group B, the tumor VEGF score, MVD and plasma VEGF level were significantly lower than Group A (P<0.05). The microscopic image of Group C showed that its number of active tumor cells and the blood capillary around tumor was significantly smaller than that of Groups A and B, and meanwhile atrophy and liquefactive necrosis were seen in local tumor. Endostar injection combined with intraperitoneal injection of cisplatin is effective in reducing tumor VEGF score and MVD of transplanted tumor tissues in rats with Lewis lung cancer to obstruct the nutrient supply of tumor cells and kill tumor cells, so that the inhibition of tumor cell proliferation and metastasis can be achieved with a remarkable effect.

Clinical observation on recombinant human endostatin combined with chemotherapy for advanced gastrointestinal cancer.

To explore the clinical efficacy and toxic and side effects of recombinant human endostatin (rh- endostatin/endostar) combined with chemotherapy in the treatment of advanced gastric cancer. A total of 70 patients with advanced gastrointestinal adenocarcioma confirmed by histopathology and/or cytological examination were divided into group A (37 patients) and group B (33 patients). Patients in group A were given intravenous drip of 15 mg endostar added into 500 mL normal saline, once every other day until the cessation of chemotherapy or patients' maximal tolerance to chemotherapy. Patients in group B received chemotherapy alone. Two groups selected the same chemotherapy regimens. FOLFIRI scheme: 90-min intravenous drip of 180 mg/m2 irinotecan, intravenous drip of 200 mg/m2 calcium folinate (CF) and 400 mg/m2 5-fluorouracil (5-Fu) on d1, and continuous intravenous pumping of 2 400 mg/m2 5-Fu for 46 h. FOLFOX4 scheme: intravenous injection of 85 mg/m2 oxaliplatin (L-OHP), 200 mg/m2 calcium folinate (CF) and 400 mg/m2 5-FU on d1 for 2 h, and then continuous intravenous pumping of 2 400 mg/m2 5-Fu for 46 h. XELOX scheme: oral administration of 1 500 mg/m2 xeloda (or tegafur 50~60 mg) in twice during d1~14 and intravenous drip of 135 mg/m2 L-OHP on d1 for 2 h. The modified FOLFOX scheme: intravenous injection of 135 mg/m2 L-OHP on d1 for 2 h, 200 mg/m2 CF and 1.0 g tegafur during d1~5. Whereas, control Group B received chemotherapy regimens which were same as Group A, but no addition of endostar. Before chemotherapy, patients were given intravenous injection of 8 mg ondansetron, intramuscular injection of 10 mg metoclopramide and 20 mg diphenhydramine for prevention of vomiting, protection of liver and stomach as well as symptomatic supportive treatment. One cycle was 21 d, 4~6 cycles in total. The efficacy was evaluated every 2 cycles. 32 patients in Group A could be evaluated, and the response rate (RR) and disease control rate (DCR) were 59.38% and 78.13%, respectively. 31 patients in Groups could be evaluated, and the RR and DCR were 32.26% and 54.84%, respectively. The differences between 2 groups were significant. The toxic effects include myelosuppression, gastrointestinal reaction, fatigue, cardiotoxicity and peripheral neurotoxicity. Preliminary observations show that endostar (once every other day) combined with chemotherapy is effective in the treatment of advanced gastrointestinal cancer, with low toxic effects, good tolerance, deserving further study.

Effect of recombinant human endostatin on the expression of c-Myc and bFGF in mouse gastric cancer cells.

The aim of this study was to observe the effects of re-combinant human endostatin on the proliferation and apoptosis of mouse gastric cancer cells, and explore some possible mechanisms of recom-binant human endostatin inhibition of cancer. A murine gastric cancer xenograft model was established. A total of 20 mice were divided into two groups (control and experimental groups). The expression of c-Myc and basic fibroblast growth factor (bFGF) was determined by reverse transcription-polymerase chain reaction, Western blotting, and immu-nohistochemical staining methods. Tumor volume was measured and a growth curve was calculated. The tumor diameter in the experimental group was significantly smaller than that in the control group after treat-ment with endostatin for 21 days. The expression levels of c-Myc and bFGF in the experimental group were significantly lower than those of the control group (P < 0.05). There was a positive correlation between the expression of c-Myc and bFGF in the experimental group. Microvessel density was significantly inhibited in the experimental group (P < 0.05). These results demonstrated that recombinant human endostatin could in-hibit tumor metastasis by inhibition of the expression of c-Myc and bFGF in gastric cancer tissue as well as by inhibition of angiogenesis.

Effect of Recombinant Human Endostatin on Angiogenesis and Treatment Response of Chemoradiation Therapy for Patients With Esophageal Squamous Carcinoma

Purpose/Objective(s)Recombinant human endostatin has been documented to be an inhibitor of tumor angiogenesis. We investigated the treatment response to anti-angiogenic therapy of recombinant human endostatin combined with chemoradiation therapy (CRT) for patients with esophageal cell squamous carcinoma (ESCC).Materials/MethodsA total of 37 ESCC patients with UICC2002 stage T1-T4 were randomly studied. Twenty-one patients were simultaneously treated with recombinant human endostatin combined with docetaxel (75 mg/m2 on day 1)/CDDP (25 mg/m2 on day 1-3)-based intensity-modulated radiation therapy (IMRT, 60-66 Gy/30-33 f/6-7 wks), and 16 patients were treated with docetaxel/CDDP-based IMRT alone. CT perfusion imaging responses of tumor blood vessels, and immunohistochemical staining of tumor microvessel density (MVD) before treatment and radiation therapy DT 20 Gy, and treatment-related complications were assessed during and after treatment.ResultsRecombinant human endostatin combined with CRT led to a marked decrease in tumor blood flow (BF) and blood volume (BV) with correction of MVD, compared to the CRT alone group (P < 0.05) was found. Our findings showed a slight improvement in overall response rate (ORR) (61.5% vs 54.5% in the CRT alone group), and no treatment-related toxicities that could be attributed specifically to recombinant human endostatin, and the common toxicities observed across the two groups are probably due to the CRT itself in the present study.ConclusionsRecombinant human endostatin combined with CRT can reduce tumor angiogenesis, and the effects of recombinant human endostatin combined with CRT were a potential improvement over that of CRT alone. Further prospective randomized control studies are needed to evaluate conclusively the clinical efficacy of this combined treatment modality for ESCC. Purpose/Objective(s)Recombinant human endostatin has been documented to be an inhibitor of tumor angiogenesis. We investigated the treatment response to anti-angiogenic therapy of recombinant human endostatin combined with chemoradiation therapy (CRT) for patients with esophageal cell squamous carcinoma (ESCC). Recombinant human endostatin has been documented to be an inhibitor of tumor angiogenesis. We investigated the treatment response to anti-angiogenic therapy of recombinant human endostatin combined with chemoradiation therapy (CRT) for patients with esophageal cell squamous carcinoma (ESCC). Materials/MethodsA total of 37 ESCC patients with UICC2002 stage T1-T4 were randomly studied. Twenty-one patients were simultaneously treated with recombinant human endostatin combined with docetaxel (75 mg/m2 on day 1)/CDDP (25 mg/m2 on day 1-3)-based intensity-modulated radiation therapy (IMRT, 60-66 Gy/30-33 f/6-7 wks), and 16 patients were treated with docetaxel/CDDP-based IMRT alone. CT perfusion imaging responses of tumor blood vessels, and immunohistochemical staining of tumor microvessel density (MVD) before treatment and radiation therapy DT 20 Gy, and treatment-related complications were assessed during and after treatment. A total of 37 ESCC patients with UICC2002 stage T1-T4 were randomly studied. Twenty-one patients were simultaneously treated with recombinant human endostatin combined with docetaxel (75 mg/m2 on day 1)/CDDP (25 mg/m2 on day 1-3)-based intensity-modulated radiation therapy (IMRT, 60-66 Gy/30-33 f/6-7 wks), and 16 patients were treated with docetaxel/CDDP-based IMRT alone. CT perfusion imaging responses of tumor blood vessels, and immunohistochemical staining of tumor microvessel density (MVD) before treatment and radiation therapy DT 20 Gy, and treatment-related complications were assessed during and after treatment. ResultsRecombinant human endostatin combined with CRT led to a marked decrease in tumor blood flow (BF) and blood volume (BV) with correction of MVD, compared to the CRT alone group (P < 0.05) was found. Our findings showed a slight improvement in overall response rate (ORR) (61.5% vs 54.5% in the CRT alone group), and no treatment-related toxicities that could be attributed specifically to recombinant human endostatin, and the common toxicities observed across the two groups are probably due to the CRT itself in the present study. Recombinant human endostatin combined with CRT led to a marked decrease in tumor blood flow (BF) and blood volume (BV) with correction of MVD, compared to the CRT alone group (P < 0.05) was found. Our findings showed a slight improvement in overall response rate (ORR) (61.5% vs 54.5% in the CRT alone group), and no treatment-related toxicities that could be attributed specifically to recombinant human endostatin, and the common toxicities observed across the two groups are probably due to the CRT itself in the present study. ConclusionsRecombinant human endostatin combined with CRT can reduce tumor angiogenesis, and the effects of recombinant human endostatin combined with CRT were a potential improvement over that of CRT alone. Further prospective randomized control studies are needed to evaluate conclusively the clinical efficacy of this combined treatment modality for ESCC. Recombinant human endostatin combined with CRT can reduce tumor angiogenesis, and the effects of recombinant human endostatin combined with CRT were a potential improvement over that of CRT alone. Further prospective randomized control studies are needed to evaluate conclusively the clinical efficacy of this combined treatment modality for ESCC.

Preliminary clinical study of recombinant human endostatin combined with radiotherapy in the treatment of brain metastases of non-small cell lung cancer and the patients suitable for this therapy

Objective To investigate the therapeutic effects of recombinant human endostatin (RHES) combined with radiotherapy on brain metastases (BM) of non-small cell lung cancer (NSCLC) and the patients suitable for this therapy. Methods Eighty patients with BM of NSCLC were randomly divided into RHES combined with radiotherapy group (combination group) and radiotherapy alone group (each group with 40 patients). The short-term effective rate, overall survival time, cerebral edema index and adverse reactions were observed and the expressions of vascular endothelial growth factor receptor 2 (VEGFR2) protein in primary lesions were detected with immunohistochemical method in all patients. Results Compared with radiotherapy alone group, brain edema was significantly relieved (t=4.9, P=0.000) and there were no marked adverse reactions in combination group. In short-term effective rate, there was no statistical significance in total population (n=80, 90% vs. 75%, χ2=3.11, P=0.07), but there was statistical significance in the patients with positive VEGFR2 (93% vs. 67.7%, χ2=6.31, P=0.012). In overall survival time, there was no statistical significance in total population (n=80, P=0.35, 95% CI: 0.25-1.30) or in the patients with positive VEGFR2 (P=0.109, 95% CI: 0.40-1.34). Conclusion Compared with radiotherapy alone, RHES combined with radiotherapy can relieve brain edema in the patients with BM of NSCLC and obtain better short-term effective rate in the patients with positive VEGFR2. Key words: Recombinant human endostatin; Radiotherapy; Vascular endothelial growth factor receptor 2; Lung cancer; Brain metastases

Effect of recombinant human endostatin on VEGF and TIMP-1 expression in hypetrophic scar of rabbit ears

Objective To introduce an accelerated solvent extraction(ASE)-HPLC-TOF/MS method for simultaneous determination of Polyphyllin Ⅰ,Polyphyllin Ⅱ,Polyphyllin Ⅵ,Polyphyllin Ⅶ,Dioscin and Gracillin in Paris Polyphylla.Methods The extraction was performed by ASE under the following conditions: extracting solvent 70% ethanol;extracting temperature 120℃;pressure 1.17 MPa(1 700 psi);and static time 6 min.The separation was carried out on a MGC18 column(3.0 mm×100 mm,3.0 μm).Elution was done with a linear gradient mobile phase system consisting of 0.1% formic acid(V/V) water and acetonitrile.The flow rate was set at 0.8 ml/min,the sample injection volume was 3 μl,the column temperature was kept constant at 25℃,nebulizer gas pressure was 275.8 kPa(40 psi),drying gas flow rate was 10 L/min,and gas temperature was 350℃.Results The calibration curves for Polyphyllin Ⅰ,Polyphyllin Ⅱ,Polyphyllin Ⅵ,Polyphyllin Ⅶ,Dioscin and Gracillin were linear within the range of 0.500 0-50.00 μg/ml(r=0.999 8),0.422 3-42.23 μg/ml(r=0.999 7),0.612 0-61.20 μg/ml(r=0.999 9),0.714 0-71.40 μg/ml(r=0.999 5),0.448 0-44.80 μg/ml(r=0.999 9) and 0.436 0-43.60 μg/ml(r=0.9997),respectively;and the average recoveries(n=6) were 98.9%,98.0%,102.5%,101.9%,103.1% and 97.9%,respectively.Conclusion The introduced method is solvent-saving and time-saving;it can also be used for high-throughput analysis and determination of steroidal saponins in Paris Polyphylla.

Anti-proliferative effect of recombinant human endostatin on synovial fibroblasts in rats with adjuvant arthritis

Rheumatoid arthritis (RA) is characterized by pronounced synovial hyperplasia resulting in pannus formation, cartilage erosion and ultimately joint destruction. Activated RA synovial fibroblasts (SFs) mediate the invasion and destruction of cartilage and bone. We previously demonstrated that recombinant human endostatin (rhEndostatin) is sufficient to induce SF apoptosis in adjuvant arthritis (AA) rats. However, the effect of this protein on SF proliferation is unknown. This study was designed to assess the inhibitory effect and mechanisms of rhEndostatin on the proliferation of cultured AA SFs. MTT assay and flow cytometric detection were performed to investigate SF proliferation and cell cycle progression, respectively. Also, the expression levels of p53, p21, cyclin D1, CDK4 and PCNA in AA SFs were detected by real-time PCR and western blotting assays. Our data revealed that AA SF proliferation was significantly inhibited by rhEndostatin in a concentration-dependent manner. In addition, rhEndostatin (50μg/ml) caused the G0/G1 cell cycle arrest of AA SFs. There were significant decreases in the expression levels of p53, p21, cyclin D1 and PCNA in AA SFs treated with rhEndostatin, and a significant increase in CDK4 expression. Collectively, our data suggest that rhEndostatin inhibits AA SF proliferation, which is preceded by cell cycle arrest at the G0/G1 phase. This is partly due to the inhibitory effect of rhEndostatin on cyclin D1 and PCNA by a p53–p21–CDK4-independent mechanism. Taken together, these findings highlight the potential use of rhEndostatin for RA treatment.

Effects of recombinant human endostatin on the expression of vascular endothelial growth factor in human gastric cancer cell line MGC-803

The present study aimed to explore the effects of recombinant human endostatin, endostar (ES), on the expression of vascular endothelial growth factor (VEGF) in the human gastric cancer cell line MGC-803. The expression of VEGF protein in MGC-803 cells was examined using immunohistochemistry. Subsequent to treatment with various concentrations of ES, the mRNA and VEGF protein expressions were determined in MGC-803 cells. A high level of VEGF protein expression was detected in MGC-803 cells. Subsequent to ES treatment, the mRNA and VEGF protein expressions were significantly decreased in MGC-803 cells (all P<0.05). In conclusion, ES is likely to inhibit the VEGF expression in MGC-803 cells.

Recombinant human endostatin inhibits adjuvant arthritis by down-regulating VEGF expression and suppression of TNF-α, IL-1β production

The aim of this study was to examine the effect of recombinant human endostatin (rhEndostatin) on adjuvant arthritis (AA) in rats and its possible mechanisms. RhEndostatin was subcutaneously administrated to AA rats after immunization. The progression of AA was assessed by the macroscopic arthritis scoring system of paws. Histological examination of the synovial tissues was examined by hematoxylin and eosin staining. The expression level of vascular endothelial growth factor (VEGF) mRNA and proteins in the synovial tissues was evaluated by realtime PCR and immunohistochemistry, respectively. Fibroblast-like synoviocytes (FLS) were isolated from synovial tissues. Cell proliferation assay was evaluateded with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. The levels of tumour necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) in culture medium was examined by radioimmno assay. RhEndostatin attenuated the severity of arthritis on both second hind paw volume and polyarthritis score, as well as improved the arthritic status histologically in AA rats. Simultaneously, rhEndostatin can inhibit the expression of VEGF in synovial tissues. The proliferation of FLS and TNF-α, IL-1β production from culture medium was significantly inhibited by rhEndostatin. Our data suggest that rhEndostatin inhibits adjuvant arthritis by down-regulating VEGF expression and suppression of TNF-α, IL-1β production.

Effects of recombinant human endostatin and its synergy with cisplatin on circulating endothelial cells and tumor vascular normalization in A549 xenograft murine model

Endostatin can normalize the tumor vasculature to some extent. However, exact length of its time window and corresponding markers for tumor vascular normalization are needed to be explored. The A549 lung adenocarcinoma xenograft murine model was treated with recombinant human endostatin (rh-endostatin) for 14days. Cisplatin was combined in different schedules. The effects of rh-endostatin on circulating endothelial cells (CECs) by flow cytometry, tumor vasculature and angiogenesis-related factors by confocal microscope and immunohistochemistry, and anti-tumor efficacy of cytotoxic drugs were observed. The activated CECs (aCECs) were increased on day 7 and decreased on day 10, and apoptotic CECs were increased on day 10. Tumor vasculature was transiently normalized with increased collagen coverage, decreased vessel permeability, intratumoral hypoxia, and microvascular density from day 7 to 10 after rh-endostatin administration. Extracellular matrix metalloproteinase inducer, vascular endothelial growth factor, matrix metalloproteinase (MMP)-2, and MMP-9 were transiently decreased by rh-endostatin from day 4 to 10, whereas the opposite effects were observed with tissue inhibitors of matrix metalloproteinase (TIMP)-1 and TIMP-2. The maximal anti-tumor effects of cisplatin were observed on administration from day 5 to 9 after rh-endostatin initial administration. Rh-endostatin could transiently normalize tumor vasculature, probably via regulation of both pro- and anti-angiogenesis factors. The synergistic efficacy of anti-angiogenesis and chemotherapy was found during "the normalization window". CEC could be a feasible blood biomarker for defining "vascular normalization window" and providing the evidence to make an optimizing combination therapeutic schedule in human tumor.