ObjectiveTo observe the expression of Rap1, guanosine triphosphate-Rap1 (GTP-Rap1), vascular endothelial growth factor (VEGF) and β-catenin in experimental choroidal neovascularization (CNV).MethodsForty-two brown Norwegian rats were randomly divided into a blank control group (7 rats) and a model group (35 rats). Both eyes were enrolled. The CNV model was established by holmium ion laser photocoagulation in the model group. At 3, 7, 14, 21, and 28 days after photocoagulation, fluorescein fundus angiography (FFA) and choroidal vascular smear were performed to observe the degree of fluorescein leakage and CNV area in rats; Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to detect the expression of Rap1, GTP-Rap1, VEGF, β-catenin and mRNA in CNV.ResultsThe results of FFA examination showed that a large disc-shaped fluorescein leaked in the photo-condensation spot 14 days after photocoagulation. Laser confocal microscopy showed that compared with 7 days after photocoagulation, CNV area increased at 14, 21, 28 days after photocoagulation, and the difference were statistically significant (t=3.725, 5.532, 3.605;P＜0.05). Western blot showed that there was no significant difference in the relative expression of Rap1 protein in CNV at different time points after photocoagulation between the two groups (P=0.156). Compared with the blank control group, the relative expression of GTP-Rap1 protein was significantly decreased, the relative expression of VEGF and β-catenin protein were significantly increased in the model group (P=0.000). The results of RT-PCR showed that there was no significant difference in the relative expression of Rap1 mRNA at different time points after photocoagulation between the two groups (P=0.645), but there were significant difference in the relative expression of β-catenin mRNA (P=0.000). At 7, 14, 21 and 28 days after photocoagulation, there were significant difference in the relative expression of GTP-Rap1 and VEGF mRNA between the two groups (P=0.000).ConclusionsThe expression of GTP-Rap1 in experimental CNV is significantly lower than that in normal rats.
Esophageal cancer is one of the common malignant tumors with high incidence and poor prognosis. Angiogenesis-related pathways play an important role in the occurrence and development of esophageal cancer. Vascular endothelial growth factor (VEGF) is the main mediator of angiogenesis. In addition to promoting angiogenesis and maintaining the survival of neovascularization, VEGF can also directly act on esophageal cancer cells and promote the occurrence and development of tumors. This article reviews the biology of VEGF and its effect on blood vessels, the expression of VEGF in esophageal cancer cells and its influencing factors, the role of VEGF in esophageal cancer cells, the immunomodulatory activity of VEGF and the clinical study of VEGF inhibitors. The purpose of this study is to provide a basis for more rational use of VEGF inhibitors in the treatment of esophageal cancer.
ObjectiveTo investigate the effects of transforming growth factor-β (TGF-β) in choroidal neovascularization (CNV) induced by laser in mice. Methods Eighty male C57BL/6J mice at the age of 6-8 weeks old were randomly divided into the normal control, photocoagulation model, photocoagulation with phosphate buffered saline (PBS control group) and photocoagulation with TGF-β receptor inhibitor groups (TGF-β receptor inhibitor group), twenty mice of each group. Fundus argon laser photocoagulation was performed in the photocoagulation model group, PBS control group and TGF-β receptor inhibitor group to induce CNV. One week, two, three and four weeks after the laser procedure, fundus fluorescein angiography (FFA) was carried out in the normal control or photocoagulation model groups to observe CNV formation dynamically. Western blot was used to analyze the expressions of TGF-β in the retina from the mice of normal control or photocoagulation model groups, and VEGF or TNF-α in the retina of normal control, PBS control or TGF-β receptor inhibitor groups. The CNV areas of each group were evaluated by using fluorescein stain on retinal pigment epithelium (RPE)/choroid flat mounts after two weeks of photocoagulation. ResultsThe FFA results showed the retinal vessels centered on the optic disc and arranged radially, while the choroidal vascular present network distribution in the normal control mice. Significant leakage of fluorescein showed discoid strong fluorophore in photocoagulation sites of retina at one week after photocoagulation. The quantitative analysis results of Western blot demonstrated that the TGF-β protein expression levels in retina of photocoagulation model mice gradually increased with time passing. The protein expression levels of TGF-β were significant differences in the photocoagulation model group comparing with the normal control group (F=13.042, P < 0.05). The protein expression levels of TNF-α (F=14.721, 17.509) and VEGF (F=18.890, 11.251) increased significantly in retina of PBS control or TGF-β receptor inhibitor groups when compared with that of normal control group at one week, two, three and four weeks after photocoagulation, and the differences were both statistically significant (P < 0.05). Compared with PBS control group, the protein levels of TNF-α and VEGF in retina from TGF-β receptor inhibitor group were significantly reduced, the differences was statistically significant (F=21.321, 16.160, P < 0.05). Two weeks after laser photocoagulation, a distinct reduction in CNV lesion size in the TGF-β receptor inhibitor group mice when compared to PBS or normal control groups, the differences was statically significant (F=4.482, P < 0.05). ConclusionTGF-β may promote CNV formation by up-regulating both TNF-α and VEGF protein expressions, the application of its specific inhibitor is able to reduce CNV progression.
Objective To evaluate an effect of the vascularendothelial growth factor (VEGF) geneactivated matrix (GAM) on repair of the sciatic nerve defect in rats. Methods The peripheral nerve extracellular matrix（ECM） was harvested by the chemical extraction from 30 SD rats. The VEGF-GAM comprised of ECM and the plasmids encoding VEGF. Thirty adult Wistar rats were made as a model of the asciatic nerve defect and were randomly divided into the following 3 groups(n=10): Group A (VEGF-GAM conduits), Group B (ECM conduits),and Group C (autografts). At 12 weeks, the rats from each groupwere subjected to an inspection for the walking tract analysis and electrophysiological and histomorphological studies.Results The VEGF DNA could be retained in GAM, promoting the transgene expressing in the sciatic nerve, and more importantly, in the axotomized neurons in the spinal cord for 12 weeks. The motor neuron recovery rate in Group A (79.13％±2.53％) was similar to that in Group C (75.26％±4.48％, Pgt;0.05), but significantly better than that in Group B (56.09％±1.89％, Plt;0.01). The number of the regenerationaxons in the distal sciatic nerve in Group A (13 463±794/mm2) was significantly lower than that in Group C (16 809±680/mm2, Plt; 0.01), but significantly higher than that in Group B (10 260±1 117/mm2,Plt;0.01). The motor nerve conduction velocity in Group A (16.44±1.65 m/s) was significantly lowerthan that in Group C (23.79±2.75 m/s, Plt;0.01), but significantly higherthan that in Group B (12.8 ±1.42 m/s, Plt;0.01). The recovery rate of thegastrocnemius muscle wet weight in Group A (71.40％±3.05%) was significantlylower than that in Group C (87.00％±1.87％,Plt;0.01), but significantly higher than that in Group B (50.00％±4.90％, Plt;0.01). The sciatic nerve function index in Group A （39.37%±4.81%） was significantly lower 〖KG6〗than that in Group C (26.27%±2.71%, Plt;0.01), but significantly higher than that in Group B (4693％±296％, Plt;0.01). Conclusion The results indicate that VEGF-GAM as a bridge can promote the functional recovery of the defected sciatic nerve in rats, but the effect is not so good as that by autografts.
ObjectiveTo explore the effect and mechanisms of bone marrow mesenchymal stem cells (BMSCs) on healing quality of acetic acid-induced gastric ulcer. MethodsForty-eight clean grade male Wistar rats were used to establish the model of gastric ulcer with acetic acid and were randomly divided into 3 groups after 3 days of modeling, 16 rats each group. After the abdominal cavity was open and stomach was pulled out, no treatment was given in group A, 150 μL phosphate buffered saline (PBS) and 150 μL BMSCs at passage 4+PBS (1×108 cells/100 μL) were injected into the gastric wall surrounding the ulcer at 5 different points in groups B and C respectively. After 10 days, the ulcer area was measured, the mucosal thickness and the number of dilated glands were tested in the regenerative mucosa by histological method. And the expression of vascular endothelial growth factor (VEGF) was detected at ulcerative margin by immunohistochemical method. ResultsThe ulcer area in group C was significantly smaller than that of groups A and B (P<0.01), but no significant difference was found between groups A and B (P>0.05). HE staining showed that group C had thicker regenerative gastric mucosa, less dilated glands, and more regular mucosal structure than groups A and B, showing significant differences in regenerative gastric mucosa thickness and dilated glands number (P<0.01), but no significant difference between groups A and B (P>0.05). Immunohistochemical staining showed that the positive expression of VEGF in the ulcer margin mucosa of group C was significantly higher than that of groups A and B. The integral absorbance (IA) value of VEGF expression in group C was significantly higher than that in groups A and B (P<0.01), but no significant difference between groups A and B (P>0.05). ConclusionBMSCs can accelerate ulcer healing by the secretion of VEGF, and improve the quality of ulcer healing.
Objective To observe the effect of vascular endothelial growth factor (VEGF) in fracture healing and to investigate the influence of VEGF and VEGF antibody in fracture healing. Methods One hundred and five rabbits were used tomake fracture model in the left radius and randomly divided into control group,VEGF group and VEGF antibody group. VEGF and VEGF antibody were used in the VEGF group and VEGF antibody group respectively, then the blood flow of the fracture ends was measured by single photon emission computed tomography (SPECT) 8,24 , 72 hours, 1, 3, 5 and 8 weeks after fracture, the X-ray films of the fracture sites were taken after 1, 3, 5 and 8 weeks to observe the fracture healing. Results The blood flow of the fracture ends in the VEGF groupincreased during aperiod from 8h to 3wk after fraction when compared with that of the control group, and no obvious difference was seen on the X-ray films between the two groups. In the VEGF antibody group, the blood flow of the fracture ends decreased obviously when compared with that of the control group. The fracture healing processwas interfered seriously and nonunion change was seen in the fracture site. Conclusion The lack of VEGF will interfere with the fracture healing process and result in nonunion in the fracture site. Administration of ectogenous VEGF may promote fracture healing by increasing the blood flow of the fracture ends.
Objective To study the expression and significance of CCR chemokine receptor-7 (CCR7) protein and vascular endothelial growth factor-D (VEGF-D) protein in the progression of breast cancer, including normal breast tissue, slight and moderate atypical hyperplasia, severe atypical hyperplasia and intraductal carcinoma in situ, as well as invasive ductal carcinoma. Methods Immunohistochemistry was used to detect the expression of CCR7 and VEGF-D protein in the nomal breast tissue (n=20), slight and moderate ductal atypical hyperplasia tissue (n=20), severe atypical hyperplasia and intraductal carcinoma in situ tissue, as well as invasive ductal breast carcinoma tissue (n=73). In addition, the D2-40 staining was also used to determine lymphatic microvessel density (LMVD). Meanwhile, the relationship between the expression of the two kinds of protein and clinicopathological factors/LMVD was analyzed by statistical analysis in breast cancer, and the correlation between expression of CCR7 protein and expression of VEGF-D protein was analyzed too. Results ①The positive rates of CCR7 protein (χ 2 =23.905，P<0.050) and VEGF-D protein (χ 2 =22.349，P<0.050) were gradually increased in the normal breast tissue group 〔CCR7 protein: 0 (0/20), VEGF-D protein: 5.0% (1/20)〕, slight and moderate atypical hyperplasia group 〔CCR7 protein: 5.0% (1/20), VEGF-D protein: 20.0% (4/20)〕, severe atypical hyperplasia and intraductal carcinoma in situ group 〔CCR7 protein: 30.0% (6/20), VEGF-D protein: 40.0% (8/20)〕, and invasive ductal carcinoma group 〔CCR7 protein: 47.9% (35/73), VEGF-D protein: 57.5% (42/73)〕. ②The LMVD value gradually increased in normal breast tissue group (2.00±1.02), slight and moderate atypical hyperplasia group (6.70± 3.48), severe atypical hyperplasia and intraductal carcinoma in situ group (9.01±2.13), as well as invasive ductal carcinoma group (16.32±4.07), there was significant difference between any 2 groups (P<0.050). ③The expressions of CCR7 protein and VEGF-D protein were correlated with clinical staging, histological grading, lymph node metastasis, and expression of human epidermal growth factor receptor-2 (HER-2) protein in patients with breast cancer (P<0.050), the higher positive rates of CCR7 and VEGF-D protein occurred in patients with higher histological grading, later clinical staging of Ⅲ+Ⅳ (compared with staging of Ⅰ+Ⅱ), lymph node metastasis (compared with no lymph node metastasis), and positive expression of HER-2 protein (compared with negative expression of HER-2 protein). The result indicated that LMVD value was related with expression of VEGF-D protein (r=0.623, P<0.010) in patients with breast cancer, but there was no correlation with expression of CCR7 protein (r=-0.303, P>0.050). Furthermore, there was weak positive correlation between expression of CCR7 protein and expression of VEGF-D protein in breast cancer (r=0.112, P<0.050). Conclusion The results strongly suggest that the expression levels of the VEGF-D protein and CCR7 protein indicate the potential of translation some extent, and they play an important role in the progression of breast cancer.
【Abstract】Objective To introduce the current studies of the role of vascular endothelial growth factorC (VEGFC) and VEGFD in lymphangiogenesis and lymph node metastasis of gastrointestinal neoplasma. Methods The related literatures in recent 5 years were reviewed. Results The growth factors VEGFC and VEGFD enhance lymphangiogenic metastasis of gastrointestinal neoplasma with the property of angiogenesis and lymphangiogenesis. In gastric adenocarcinoma, VEGFC mRNA and tissue protein expression correlate with lymphatic invasion, lymph node metastasis, venous invasion and reduced 5year survival rates. The role of VEGFC in esophageal squamous cancer and colorectal cancer and VEGFD in colorectal cancer is not certain, with conflicting reports in the published literatures.Conclusion The VEGFC, VEGFD/VEGFR3 signal pathway may become the ideal target for inhibition of tumor proliferation and metastases, antilymphangiogenesis therapy may be a novel potential strategy in tumor biological therapy.
Objective To investigate the expression of pigment epitheliumderived factor (PEDF) and vascular endothelial growth factor (VEGF) in choroidal melanoma. Methods The expression of VEGF and PEDF protein in fifty-eight cases of paraffinembeded choroidal melanoma samples was measured by immunohistochemistry, the expression of PEDF mRNA in thirtynine choroidal melanoma samples was assayed by in situ hybridization. Results PEDF protein was detected in 13/58 cases (22.4%) of choroidal melanoma, the positive rate in nonsclerainvasion group (12/38, 31.6%) was higher (Plt;0.05) than that in sclerainvasion group (1/20, 5%). VEGF protein was detected in 43/58 cases (64%) of choroidal melanoma, the positive rate in nonsclerainvasion group (25/38, 65.8%) was lower (Plt;0.05) than that in sclerainvasion group (18/20, 90%). The expression of PEDF mRNA was detected in 18/39(46.2) cases, the positive rate in nonsclerainvasion group was higher (Plt;0.05) than that in sclerainvasion group. Conclusions Imbalanced expression of VEGF and PEDF in choroidal melanoma may play a key role in the angiogenesis, tumor progression and metastasis.
ObjectiveTo investigate the relationship between vascular endothelial growth factor receptor-3 (VEGFR-3) and clinical pathology of gastric carcinoma(GC).MethodsThe expression of VEGFR-3 in 80 GCs and 20 gastric benign tissues (GBT) was detected by immunohistochemistry(SP), by which the density of lymphatic vessels (DLV) was calculated. ResultsThe DLV in GC was （5.800 0±2.318 9）/×200, in GBT （2.380 0±0.462 9）/×200(P=0.000); in GC with lymph node metastasis （6.948 3±1.583 1）/×200, without lymph node metastasis （2.772 7±0.428 9）/×200 （P=0.000）. In poorly differentiated type group, DLV was （7.681 8±0.982 9）/×200, higher than that in moderately and highly differentiated type group 〔(3.500 0±1.028 2）/×200, P=0.000〕. DLV in pTNM Ⅰ+Ⅱ was （4.291 7±1.688 0）/×200, in Ⅲ+Ⅳ （8.062 5±0.759 4）/×200 （P=0.000）.ConclusionDLV shows positive relations with pTNM stage, differentiation and lymph node metastasis of GC.