Objective To investigate the serum levels of endostatin and vascular endothelial growth factor ( VEGF) at different therapy stages of mouse Lewis lung carcinoma, and elucidate the relation to the progress and prognosis of tumor. Methods Forty-four Lewis lung carcinoma-bearing C57BL/6 mice were randomly divided into 4 groups, ie. a non-therapy group, a chemotherapy group, a gene therapy group, and a combination therapy group ( chemotherapy plus gene therapy) . Eleven healthy mice were included as normal control group. Serum was collected on the 0th, 5th, 19th day after therapy for measurement of endostatin and VEGF by ELISA. The correlations were analysed between endostatin and VEGF levels in each group. Results ( 1) The serum endostatin levels had no significant difference in all groups on the 0th day,but increased significantly on the 5th day in the gene and combination therapy groups than those in other three groups ( all P lt;0. 01) . Then the endostatin level decreased on the 19th day in the gene and combination therapy groups, but still higher than those in the chemotherapy group and the normal group. ( 2 ) On the contrary, serum VEGF levels of the gene and combination therapy groups decreased significantly on the 5th day and increased little on the 19th day, which were both significant lower than those in chemotherapy group on the 5th and 19th day( all P lt; 0. 05) . There were significant diferences between the three therapy groups and the non-therapy group( all P lt;0. 05) . ( 3) Negative correlations between VEGF and endostatin levels were revealed in the gene and combination therapy groups ( r = - 0. 77 and - 0. 761 respectively) .Correlation was not found in the non-therapy and chemotherapy groups. Conclusion The serum levels of endostatin and VEGF might be used as monitoring indices of antiangiogenesis therapy.
Objective To construct the recombined DNA pcDNA3.1-hBMP-2 and transfect into human marrow stromal stem cells (MSCs) in vitro, and to explore theeffects of transfection on cellular proliferation and expression of vascular endothelial growth factor (VEGF). Methods The expression of human bone morphogenetic protein 2(hBMP-2) in these cells after transfection was determined by in situ hybridization and immunohistochemical analysis and Western blot analysis. The changes of cell proliferation were observed by flow cytometry. The effects of BMP-2 gene transfection on expression of VEGF in the cells were analyzed by in situ hybridization of VEGF cDNA probe. Results Stable expressionof hBMP-2 in pcDNA3.1-hBMP-2 transfected MSCs was confirmed in the levels of mRNA and protein.Cellular proportion in S period increased, which indicated that the synthesis of cell DNA increased. The expression of VEGF in the cells increased obviously. Conclusion With the help of lipofectamine, the pcDNA3.1-hBMP-2 were transfected into human MSCs successfully. hBMP-2 plays an important role in promoting cellular proliferation and vascular generation during bone repair.
【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 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 explore a new method of treating early avascular necrosis of femoral head (AVNFH). Methods Sixty-nine New Zealand adult rabbitswith a mean weight of 2.8 kg after AVNFH presenting were randomly divided into three groups. In group A, deproteinized bone(DPB) combined with the recombinant plasmid pcDNA3.1/vascular endothelial growth factor 165(VEGF165) was implanted in the drilled channel of the necrotic femoral head. In group B, only DPB was implanted. In group C, channel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained 3 days, 1, 2, 4, 8 and 16 weeks after operation. The expression of VEGF165 was examined by RT-PCR, Western blot and immunohistochemical techniques. X-ray testedbone formation generally. Angiogenesis and repair of the femoral head were observed by histological and histomorphometric analysis. Results In group A, the expressions of VEGF165 mRNA and protein were detected 3 days postoperatively, reached apex 1 week and lasted more than 3 weeks after implantation. The ratios of IOD of collagen type Ⅰ were 0.29±0.11, 0.55±0.13 and 0.67±0.10 IOD/μm2 respectively at 2, 4 and 8 weeks postoperatively and the ratios of IOD of new capillary vessels were 0.33±0.10and 0.57±0.16 IOD/μm2 respectively at 2, 4 weeks postoperatively in group A, showing statistically significant difference (Plt;0.01) when compared with groups B and D. X-ray test indicated much bone callus formed early. Conclusion Transfection of the VEGF165 gene can enhance local angiogenesis at early stage andDPBVEGF165 compound can improve bone formation. Deproteinized bone combined with VEGF165 gene provides a potential method for therapy of osteonecrosis.
ObjectiveTo evaluate the effects of nerve growth factor (NGF) on angiogenesis and skeletal muscle fiber remodeling in ischemic hindlimbs, and study the relationship of NGF and vascular endothelial growth factor (VEGF) to angiogenesis. MethodsEighteen mice were randomly allocated to normal control group (n=6), blank control group (n=6), and NGF gene transfection group (n=6). The left hindlimb ischemia model was established by ligating the femoral artery. NGF plasmid (125μg) was injected into the mouse ischemic gastrocnemius in the NGF gene transfection group. The same volume of normal saline (200μL) was injected into the mouse ischemic gastrocnemius in the blank control group. The gastrocnemius of left hindlimb was harvested under the condition of peritoneal cavity anesthesia on the 21th day after operation, and then the mice were sacrificed. The gastrocnemius of three groups were tested by hematoxylin-eosin staining, proliferating cell nuclear antigen (PCNA) and CD34 were determined by immunohistochemistry staining. Skeletal muscle fiber type was tested by myosin ATPase staining. NGF and VEGF protein expression were detected by enzyme linked immunosorbent assay. ResultsOn the 21th day after surgery, compared with the blank control group, the skeletal muscle atrophy degree was weaker, the functional assessment score was significantly lower (P < 0.05), the endothelial cell proliferation index, capillary density, the typeⅠskeletal muscle fiber proportion, NGF and VEGF expression were significantly higher (P < 0.05) in the NGF gene transfection group. ConclusionsNGF gene transfection could promote NGF and VEGF expression and angiogenesis in ischemic hindlimbs, and induce typeⅠskeletal muscle fibers formation in ischemic hindlimbs. The molecular regulation mechanism still needs to be further studied.
Objective To evaluate the effect of vascular endothelial growth factor (VEGF) on tumor angiogenesis, and its usage in tumor therapy.Methods The recent literatures about VEGF and angiogenesis were reviewed and analyzed. The advances of VEGF study were summarized. The effects of anti-angiogenesis in tumor biological therapy were introduced.Results Angiogenesis had been identified as an important factor for promoting tumor growth. VEGF was a basic and pivotal factor in tumor angiogenesis. The anti-angiogenesis treatments aimed at VEGF, including the applications of VEGF inhibitor and gene therapy of adenovirus medium, had got great progress. Conclusion VEGF is a leading factor of tumor angiogenesis, the anti-angiogenesis therapy aimed at VEGF has probably provided a new chance to malignant tumor treatment.
For research the relationship between the expression of vascular endothelial growth factor (VEGF) and the cell proliferation. The expression of VEGF was evaluated while the cell cycle of hepatoma cell line Hep G2, which was synchronized at G0 phase with serum deprivation, and reinitiated with TPA and blocked with antisense oligonucleotides of c-jun. Results: Hep G2 cell did not express VEGF at G0 phase. TPA could induce the expression of VEGF as well as initiation of cell cycle. The antisense oligonucleotides of c-jun could prohibit the expression of VEGF and arrest the cell cycle at G0 phase. Conclusion: The fact that the expression of VEGF accompanies the initiation of cell cycle suggests that they be regulated by the same singnal pathway, the expression of VEGF may be a marker indicating the proliferation of hepatoma cells.
ObjectiveTo observe the effects of hydroxysafflor yellow A (HSYA) on microvessel density (MVD) of mice transplanted Lewis lung cancer and mRNA expression of vascular endothelial growth factor (VEGF) so as to explore the tumor-inhibiting mechanism of HSYA. MethodsSixty tumor-bearing C57/BL mice were randomly divided into five groups, with 12 mice in each group, namely a control group, a cyclophosphamide (CTX) group (25mg/kg), a large dose HSYA group (112mg/L), a medium dose HSYA group (56mg/L), and a small dose HSYA group (28mg/L). These different drugs were administered by intraperitoneal injection. The mice were sacrificed 22 days after the treatment. Tumor tissues were sampled and examined by immunohistochemical method and quantitative real-time PCR to detect the expression of MVD and VEGF mRNA. ResultsThe MVD of the medium and small dose HSYA groups and CTX group were 30.01±3.12, 22.56±2.11 and 16.21±2.40, respectively, which were significantly lower than 41.10±2.93 of the control group and 37.66±3.04 of the large dose HSYA group (χ2=2.82, P=0.010;χ2=3.16, P=0.007;χ2=4.58, P=0.000) and (χ2=1.98, χ2=0.038;χ2=2.45, P=0.016;χ2=3.82, P=0.001). The difference in VEGF amplified fluorescence expression threshold between the HSYA groups and the control group was not significant. However, after amplification, the expression of VEGF mRNA in the small dose HSYA group was only 0.43±0.16, which was obviously lower than 0.82±0.06 in the control group (F=0.77, P=0.038). ConclusionHSYA can significantly reduce MVD in mice transplanted Lewis lung cancer and down-regulate expression of VEGF mRNA to achieve tumor-inhibiting effect.
Objective To research the effects of vascular endothelial growth factor (VEGF) on the survival of reverse flow axial skin flaps. Methods A 8 cm×2 cm full thickness transverse dorsal flap based on right deep circumflex iliacartery was elevated in 20 SpragueDawley rats, which length crossing midline was 4.0 cm. The rats were randomized into two groups:experimental group (n=10), subcutaneous VEGF injections into the flap (200 ng, 200 μl) after flap elevation; controlgroup (n=10), subcutaneous saline injections into the flap (200 μl) after flap elevation. The flap was immediately sutured to its recipient beds then the injection was executed. Seven days after operation, the survival area of flaps and density of vessels were observed and measured, meanwhile its histological representation of the flaps was examined. Results After 7 days of recovery, the mean survival area of flaps was 15.55±0.27 cm2 in experimental group and 13.42±0.57 cm2 in control group. The difference was significant between experimental group and control group (P＜0.01). The mean vessel density of flaps was 21.00±3.16 in control group and 34.40±3.75 in experimental group. The difference was significant between experimental group and control group (P＜0.01). Histological analysis demonstrated that a qualitatively greater amount of granulation tissue, regular collagen fiber and a lot of fibrillated cells were observed in experimental group. Erythrocytes were leaked outfrom vessels, and inflammatory cells were observed around in control group. Conclusion In early survival of flaps, the VEGF can improve the survival of areverse flow axial skin flap through improving angiogenesis and increasing the perfusion of vessel. It is an effective method of improving the survival of reverse flow axial skin flaps that VEGF is fully injected in subcutaneous flaps by single, when flaps are elevated.