【Abstract】Objective To evaluate the status of vascular endothelial growth factor (VEGF) expression in breast carcinoma and benign disease and define the relationship with age,menopause, tumor size，clinical stage，distant metastasis and lymph node metastasis. Methods Seventy cases of invasive ductal breast carcinomas,30 benign breast diseases and 7 adjacent nonneoplastic specimens were assessed for VEGF protein expression by immunohistochemistry LSAB method. Results VEGF were expressed more frequently in breast cancer than in benign diseases.VEGF was significantly correlated with axillary lymph node metastasis and distant metastasis,whereas no statistical correlation with other factors. Conclusion VEGF status has certain value to make differential diagnosis between malignant and benign breast diseases and predict the possibilities of distant and lymph node metastasis.
Objective To investigate the effects of adenovirus-mediated melanoma differentiation-associated gene-7 (mda-7)/IL-24 and/or adriamycin (ADM) on transplanted human hepatoma in nude mice and to explore a new way for hepatoma gene therapy combined with chemotherapy. Methods The recombinant adenovirus vector carrying Ad.mda-7 was constructed; Ad.mda-7 and/or ADM were injected into the tumor-bearing mice. Their effects on the growth of the tumor and the survival time of the mice were observed. The expressions of VEGF and TGF-β1 were detected by an immunohistochemistry method. Results Ad.mda-7 was constructed and expressed in vivo successfully. Compared with other three groups 〔control group (43.4±1.67) d, ADM group (64.2±4.14) d, Ad.mda-7 group (61.4±1.67) d〕, the mice treated with Ad.mda-7 combined with ADM had longer average survival time 〔(83.8±4.82) d, P＜0.01〕; the average size of tumor treated with Ad.mda-7 combined with ADM diminished significantly compared with that treated with ADM or Ad.mda-7 separately (P＜0.01). VEGF and TGF-β1 expressions of Ad.mda-7 group were (56.2±7.7)%, (35.2±4.5)%, respectively, and were lower than those in ADM group (VEGF: P＜0.05; TGF-β1: P＜0.01). VEGF expression of Ad.mda-7+ADM group was (37.3±5.0)%, and was significantly lower than that in other three groups (P＜0.01). TGF-β1 expression of Ad.mda-7+ADM group was (31.2±3.1)% and significantly lower than that in control group and ADM group (P＜0.01), however, there was no significant difference compared with Ad.mda-7 group (Pgt;0.05). Conclusion Ad.mda-7 combined with ADM has b antitumor potency and synergistic effects and suppresses the growth of human HCC xenograft in nude mice, possibly by inducing the apoptosis of hepatoma cell lines and suppressing tumor angiogenesis.
Objective To investigate the effect of local delayed releasing vascular endothelial growth factor (VEGF) on accelerating healing of intestinal anastomotic stoma. Methods An intra-abdominal infection modal of rabbit was established by artificial appendix perforation, and excision and anastomosis of terminal ileum were subsequently performed after 12 h. The animals were divided into four groups (32 for each group) with different reagents on anastomotic surface: control group, fibrin glue group (FG group), VEGF group, and VEGF＋FG group. The incidence of stomal leak, anastomosic bursting pressure, hydroxyproline content, and expression of VEGF in cured stoma tissue were measured respectively on day 3, 5, 7 and 14 after operation. Results The total incidence rate of leakage was lower in FG group and VEGF group than that in control group, but there was no statistical significance （Pgt;0.05）. The incidence rate was significantly lower in FG＋VEGF group than that in control group （Plt;0.05）. On day 14 postoperatively, the bursting pressure of anastomotic stoma, hydroxyproline content, and positive cell expression rate of VEGF protein (except VEGF group) were significantly increased in FG＋VEGF group than those in other three groups （Plt;0.05, Plt;0.01）. Conclusion Local delayed release of VEGF by fibrin glue can improve the healing of intestinal anastomotic stoma and reduce the incidence of stomal leak.
【Abstract】Objective To observe the changeable expressions of vascular endothelial growth factor (VEGF) and integrin β3 during the angiogenetic process of granulation tissue. Methods mRNA and protein of VEGF and integrin β3 in human normal subcutaneous tissue, proliferative granulation tissue and mature granulation tissue were observed by RT-PCR and immunohistochemistry staining. Results The expressions VEGF and integrin β3 were low in normal subcutaneous tissue and were much higher in proliferative granulation tissue. When the granulation tissue was mature, the expression was decreased again. Conclusion VEGF and integrin β3 are important regulating factors in ngiogenesis.
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 understand the features of lymphatic vessel, and to summarize the foundation and mechanism of the promotion and inhibition of tumor lymphangiogenesis recorded on the current studies of animal experiments and clinical researches. Methods The related literatures of the structural features of lymphatic vessel, lymphatic endothelial molecular markers, the origin of lymphatic tumors, the molecular mechanisms and regulatory factors were reviewed, and the relationship between tumor lymphangiogenesis and lymphatic metastasis, the treatment targeting at the formation of the anti-tumor lymphatic vessel and its existing problems were also analyzed. Results Hyperplasia of lymphatic vessels occurred during the process of tumor formation and progression. The structural features of the lymphatic vessels in the tumor were conducive to tumor lymphatic metastasis. In recent years, methods of anti-lymphangiogenesis and inhibition of tumor lymphatic metastasis had achieved considerable success in animal experiments. However, there were still a lot of problems need to be solved. Conclusion Tumor lymphangiogenesis has a significantly positive correlation between tumor lymphatic metastasis and patients’ prognosis, which may indicate that treatment against the formation of tumor lymphatic vessel maybe effective.
Objective To detect the expressions of osteopontin （OPN）， breast tumor kinase （Brk）， and vascular endothelial growth factor （VEGF） in the breast cancer tissue， the adjacent （2cm） normal breast tissue， and the distal（＞5cm） normal breast tissue， and analyze their clinical significances. Method The immunohistochemical method was used to detect the expressions of OPN， Brk， and VEGF in the breast cancer tissue， the adjacent （2cm） normal breast tissue， and the distal （＞5cm） normal breast tissue from 40 cases of breast cancer. Results ① The expressions of OPN，Brk， and VEGF in the breast cancer tissue were significantly higher than those of the adjacent （2cm） normal breast tissue and the distal （＞5cm） normal breast tissue （P＜0.01）， the expression of Brk in the adjacent （2cm） normal breast tissue was significantly higher than that of the distal （＞5 cm） normal breast tissue （P＜0.05）. ② In the breast cancer tissue， the OPN and Brk protein expressions were not associated with age， tumor diameter， and histological grade （P＞0.05），were associated with lymph node metastasis and TNM stage （P＜0.05）. The VEGF protein expression was not associated with age and tumor diameter （P＞0.05）， but was associated with histological grade， lymph node metastasis， and TNM staging （P＜0.05）. ③ In the breast cancer tissue， OPN， Brk， and VEGF had positive correlation with each other （P＜0.05）， but not in the adjacent （2cm） normal breast tissue and the distal （＞5 cm） normal breast tissue （P＞0.05）. Conclusions The expressions of OPN and Brk from the same signal pathway increase by turns in the distal （＞5 cm） normal breast tissue， adjacent （2cm） normal breast tissue， and breast cancer tissue. OPN induced the adhesion and migration of endothelial cells to accelerate vascular repair through VEGF and Brk has correlation with the progress of tumor invasion and metastasis through participating in tumor vascularization.
Objective To investigate the effect of Adenovirus-mediated averse vascular endothelial growth factor165（Ad-aVEGF165）on the growth of human melanoma cells(A375) in vivo and in vitro.Methods In vitro,the 100 multiplicity of infection of Aadenovirus-mediated green fluorescent protein（Ad-GFP）and Ad-aVEGF165 were transfected into human endothelium cell of vessel 304（ECV 304） and A 375. ECV 304 cells were divided into 3 groups: A 375 group, AdGFP group and AdaVEGF 165group. A375cells were also divided into 3 groups:1640 group, Ad-GFP group and AdaVEGF165 group. Their effects were analyzed by proliferation assay, cell cycle, and VEGF expression. In vivo,A375cells were injected into the axilla of the nude mouse. When the tumor formed, they were transplanted into another 15 mice. After treatment, the tumor was excised for naked eye observation, HE observation and microvascular density(MVD) counting. Results The cell supernatant fluid of A 375 group and AdGFP group could stimulate ECV304 cell growth，butthat of AdaVEGF165 group could inhibit the growth of ECV304 cell.All the A375cells in 3 groups had the proliferation trend, showing no statistically significant difference（Pgt;0.05）. ECV 304 cell proliferation index(PI) in Ad-aVEGF165group reduced（Plt;0.05）. There was no statistically significant difference（Pgt;0.05） in the PI of A 375 cell. The A 375cell integral optical densities were 234.41±13.8 in 1640 group， 222.73±3.67 in AdGFP group and 180.84±6.34 in Ad-aVEGF165group. The tumor volume in Ad-aVEGF165 group was smaller than that in Ad-GFP group and PBS group at 2 weeks after operation, the trend became much obvious with the time delay. AdaVEGF165 brought to much tissue necrosis under HE stain. The MVD of PBS group, Ad-GFP group and Ad-aVEGF165group were 65 10/view,52±11/view and 30±6/view, respectively. Conclusion In Vitro, Ad-VEGF 165gene could inhibited ECV304 cells’ growth by weakening VEGF expression of A 375cells. In vivo, Ad-aVEGF 165could inhibit the growth of human melanoma from blockinmicrovascular.
Objective To study the expression of inducible nitric oxide synthase (iNOS)，endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) in human gastric cancer and their relationship with tumor angiogenesis and to investigate the interaction of NOS and VEGF in gastric cancer. Methods The expression and distribution of VEGF, iNOS and eNOS in 34 gastric cancer specimens were detected with immunohistochemistry. Microvessel density (MVD) was counted with FⅧRAg immune specific staining. Results The expression rates of iNOS, eNOS and VEGF in 34 gastric cancers were 73.5%, 82.4% and 91.2% respectively. The expression of VEGF had a significant positive relation with iNOS, but not with eNOS. The MVDs of VEGF or iNOS positive gastric cancers were obviously higher than those of VEGF or iNOS negative gastric cancers. There was no significant difference between the MVDs of eNOS positive gastric cancers and eNOS negative ones. Conclusion MVD increases with increase of expression of VEGF and iNOS in gastric cancer. It is indicated that VEGF and iNOS can promote gastric cancer angiogenesis. VEGF and iNOS have a significant positive correlation, which suggests that in human gastric cancer, iNOS plays an important role in the production and action of VEGF.
ObjectiveTo analyze the regulative rule of mRNA of vascular endothelial growth factor (VEGF) in mice with oxygen-induced retinopathy, and to elucidate the possible mechanism of occurrence of neovascularization in retinopathy of prematurity (ROP).MethodsSixty 7-day-old C57BL/6J mice were divided into oxygen-induced retinopathy group and control group. In oxygen-induced retinopathy group, 36 mice were exposed to 75% oxygen for 5 days and then to room air for 5 days; in control group, 24 mice were raised in room air. Vascular perfusion of fluorescein and retinal stretched preparation were used to observe the morphologic changes of retinal vessels. Reversal transcriptionpolymerase chain reaction (RT-PCR) was used to observe changes of VEGF mRNA in each group. ResultsIn oxygen-induced retinopathy group, the morphologic characteristics of retinal vessels were the unperfused area at the center of superficial and deepseated vessels, and the neovascularization appeared at mid-peripheral retina after 2 days in relative hypoxia condition. The results of RT-PCR showed space-time corresponding relation between expression of VEGF and neovascularization, which meant that the transcription of VEGF mRNA decreased in hyperxia conditionand increased in relative hypoxia condition. ConclusionHypoxia is the main reason of occurrence of retinal neovascularization; increased expression of VEGF caused by relative hypoxia after hyperxia might be effective in reducing the occurrence of neovascularization in ROP.(Chin J Ocul Fundus Dis, 2005,21:292-295)
