[Effect of vascular endothelial growth factor 165 gene transfection on repair of bone defect: experiment with rabbits].
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To investigate the influence of vascular endothelial growth factor (VEGF) 165 gene transfection on the repair of bone defect.38 New Zealand rabbits underwent resection of a segment 1 cm in length in bilateral radii filled with absorbable gelatin sponge. Dilated solution of the plasmid pcDNA3.1/VEGF165 was injected into the bone defect of one side and normal saline was injected into the contralateral bone defect. 1, 2, 4, 6, 8, and 12 weeks later X ray examination was conducted to observe the repair of bone defect, and then 5 rabbits were killed at each time points to take out the bone defects. HE staining was used to observe the bone repair. The levels of microvessel density (MVD) 1 and 2 weeks after the operation were observed. RT-PCR was used to detect the mRNA expression of VEGF in the bone defect. Based on the results of RT-PCR the tissue mRNA expression of VEGF65 was detected by real-time quantitative polymerase chain reaction (RQ-PCR).X-ray examination showed that there was no significant difference in the wound healing between the two group 1 week after the operation in all rabbits. Some callus could be seen in the experimental group 2 weeks after. Twelve weeks after the operation the reconstruction of bone cortex was completed. Similar process occurred in the control sides but more lately. The MVD level 7 days after of the experimental group was 47.0 +/- 7.5, significantly higher than that of the control group (42.2 +/- 6.4, t = 2.4519, P = 0.0179), and the MVD level 14 days after of the experiment group was 69.1 +/- 5.4, significantly higher than that of the control group (56.1 +/- 6.1, t = 8.0347, P = 0.0000). In the experimental group the mRNA expression amounts of VEGF165 could be found 1 week after, gradually increased and peaked 3 weeks after, then decreased, and became stable 6 weeks after. The mRNA expression amounts of VEGF165 in the control group were lower than those of the experimental group.Local application of PcDNA3.1/VEGF(165) vector promotes the expression of VEGF165, and enhances the quantity of the angiogenesis, extra cellular matrix and healing of bone defect.Cite
Angiogenesis and bone formation are vital for fracture healing. Nerve growth factor (NGF) not only promotes neuronal survival but also enhances the proliferation and differentiation of osteoblasts. Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis. However, the potential correlation of NGF and VEGF levels with fracture healing in patients with traumatic brain injury (TBI) remains unclear.This study enrolled 22 patients with clavicle fracture and concomitant TBI (CFT group) and 25 patients with clavicle fracture alone (CF group). Serum NGF levels were measured with ELISA. The expressions of NGF, VEGF, and CD31 in callus tissues were measured with immunohistochemistry.The fracture healing time in CFT group (82.22±13.61 days) was significantly shorter than that in CF group (127±25.05 days; P<0.001). The expression of CD31, marker of blood vessels, in callus tissues of CFT group was higher compared with that of CF group. Serum NGF levels and the expression of NGF in callus tissues of CFT group were higher than those in CF group (P<0.01). The expressions of CD31, NGF, and VEGF are correlated with shorter fracture healing time.The formation of blood vessels was increased in CFT group compared with CF group. NGF and VEGF levels were higher in CFT group than in CF group and correlated with shorter fracture healing time. Accelerated fracture healing in patients with TBI may be due to NGF- and VEGF-mediated angiogenesis at the fracture site.
CD31
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Parthenogenesis
Vasculogenesis
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Vascular endothelial growth factor (VEGF)-induced angiogenesis is involved in the etiology of some cardiovascular diseases. The soluble form of VEGF receptor, FLT-1 (sFLT-1), is a potent antagonist of VEGF. Therefore, we investigated whether transfection with the sFLT-1 gene could inhibit VEGF-induced angiogenesis. Human embryonic kidney (HEK)-293 cells were transfected with plasmids containing VEGF and sFLT-1 (pCMV-VEGF and pCMV-sFLT-1) by the calcium-phosphate co-precipitation method. VEGF- and/or sFLT-1-transfected HEK-293 cells were incubated for 24 h, and then conditioned medium was collected. The effects of conditioned medium on angiogenesis were tested by incorporation of [3H]thymidine into human umbilical vein endothelial cells (HUVECs). Expression of VEGF protein was determined by Western blotting. The conditioned medium from sFLT-1 gene-transfected HEK-293 cells significantly inhibited recombinant VEGF-induced increase in [3H]thymidine incorporation by HUVECs. VEGF gene-transfected HEK-293 cells secreted VEGF protein into conditioned medium. This conditioned medium increased [3H]thymidine incorporation by HUVECs, which was significantly inhibited by co-transfection of sFLT-1 gene with VEGF gene. These observations suggested that sFLT-1 gene transfer could inhibit VEGF-induced DNA synthesis of vascular endothelial cells.
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Objective To explore the expression and its implication of angiogenesis and invasiveness related factor in primary and recurrent glioma. Methods Expressions of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected by immunohistochemical technique. The morphological characteristics of ultrastructure of glioma were observed by transmission electron microscope (TEM). Results The expressions of VEGF, MMP-2 and MMP-9 varied in different grades of primary glioma. With the elevation of the malignant degree of the primary glioma, positive staining rates of VEGF, MMP-2 and MMP-9 increased significantly. The expression of VEGF correlated with both MMP-2 and MMP-9 expression. Compared with the primary glioma, the immunoreactivities of VEGF, MMP-2 and MMP-9 in recurrent glioma increased, especially in those with more severe malignancy. Under transmission electron microscope, endothelial cells markedly proliferated and protruded from the deficiency of basemembrane, concomitantly with edema of the extracapillary gap, plasma extravasation as well as some small worm-eaten caverns in the basemembrane.Conclusion VEGF, MMP-2 and MMP-9 play important roles in glioma angiogenesis and invasiveness.Inhibition of their expressions may be a useful therapy to glioma.
Key words:
Glioma; Vascular endothelial growth factor A; Matrix metalloproteinases; Microscope, electron, transmission
Extravasation
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目的将学习 4 polyamidoamine/vascular endothelial 增长系数反感觉 oligodeoxynucleotide (G4PAMAM/VEGFASODN ) 在脉管的 endothelial 增长系数(VEGF ) 和它乳癌房间并且在脉管的 endothelial 房间的抑制上的 mRNA 的表情上加重的产生的效果。我们检验了 G4PAMAM/VEGFASODN 化合物和它的 pH 稳定性的形态学的方法在 vitro transfection 效率和毒性，并且 VEGF 和它的 mRNA 的表情。甲基 thiazolyl tetrazolium 试金被用来在脉管的 endothelial 房间上检测混合物的禁止的功能。结果混合物是在直径的大约 10 nm 并且是同类地网状的。从 pH 5 ～ 10，它显示出相当一个缓冲能力。在 1:40 的费用比率的 48-h transfection 率是 98.76% ，比脂肪的一些组(P<0.05 ) 的显著地高。任何一个都没在房间上 transfection 产品显示出明显的毒性。在 G4PAMAM/VEGFASODN transfection 以后的 VEGF 蛋白质和它的 mRNA 的表情显著地减少了。有低毒性，高安全，和高 transfection 的结论评价， G4PAMAM/VEGFASODN 能是有希望的基因向量。明确地，它高效地禁止 VEGF 基因表示，放一个基础因为进一步在 vivo 动物学习。
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Fractures require adequate stability and blood supply to heal. The vascular supply to long bones is compromised in a fracture, and the ability to heal hinges on the ability of new blood vessels to proliferate from surrounding vessels in a process known as angiogenesis. This process is largely driven by the growth factor, vascular endothelial growth factor (VEGF), whose levels are increased locally and systemically during fracture healing. VEGF is involved in many steps throughout the fracture healing cascade, from initially being concentrated in fracture hematoma, to the promotion of bone turnover during the final remodeling phase. This article reviews the current literature surrounding the role of VEGF and other growth factors in reestablishing vascular supply to fractured bone, as well as medications and surgical techniques that may inhibit this process.
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Objective To investigate the effect of tumor susceptibility gene 101 (TSG101) on angiogenesis of gastric cancer SGC7901 cells.Methods Cells were divided into the non-transfection group,the empty plasmid transfection group and TSG101 eukaryotic expression plasmid transfection group (the TSG101 transfection group).Then we examined the relationship between TSG101 expression and tumor angiogenesis in vivo and in vitro experiment respectively.To detect the expression of vascular endothelial growth factor (VEGF),vascular endothelial growth factor receptor (VEGFR) and the angiogenesis.Resuits (1) It was showed that the angiogenesis number of HUVEC and the expression levels of VEGF in the TSG101 transfection group was significantly higher than that of the empty plasmid transfection group and the non-transfection group in vitro experiment (19.50 ± 3.02 vs.11.50 ± 2.51,11.34 ± 1.63 and 514.80 ± 18.16 vs.306.58 ± 20.86,305.69 ± 29.93,all P ＜ 0.05).(2) It was showed the microvascular density in tumor tissue of nude mice and the serum VEGF concentration of the TSG101 transfection group was significantly higher than that of the empty plasmid transfection group and the non-transfection group in vivo experiment [11.16±2.31 vs.5.33 ±1.63,4.66 ±1.36 and (340.64±19.15) ng/L vs.(219.34 ± 13.56),(206.10 ± 25.85) ng/L,all P ＜ 0.05].The expressions of VEGF and VEGFR of the TSG101 transfection grou p (0.548 ± 0.032,0.345 ± 0.042) of tumor tissue in nude mice were significantly higher than that of the non-transfection group (0.350 ± 0.034,0.203 ± 0.030) and the empty plasmid transfection group (0.327 ± 0.020,0.206 ± 0.036,P ＜ 0.05).Conclusion The high-expression of TSG101 in SGC7901 cells may promote angiogenesis of gastric cancer by up-regulated expression of VEGF.
Key words:
Tumor susceptibility gene 101 ; Angiogenesis; Vascular endothelial growth factor; Gastric cancer
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Vascular endothelial growth factor (VEGF) is a potent paracrine angiogenic factor involved in angiogenesis. We determined whether antisense VEGF transfection can suppress angiogenic activity of a human squamous cell carcinoma of the head and neck (SCCHN) cell line.Human SCCHN cell lines were screened for VEGF secretion by ELISA. The highest VEGF secreting cell line was transfected with an antisense VEGF vector. Endothelial cell migration assays were performed using the conditioned medium from the transfected clones. Tumorigenicity assays of the transfectants in nude mice were also performed.Antisense VEGF expression exhibited a 20-fold inhibition of VEGF secretion. The addition of conditioned medium from the antisense clones resulted in 50% reduction of endothelial migration. There was no effect on in vivo tumorigenicity.Antisense VEGF transfection effectively down-regulated VEGF secretion from SCCHN cells that had high VEGF secretion. Targeting VEGF expression may be useful for suppressing angiogenesis in head and neck cancer.
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To explore a new method for the therapy of avascular necrosis of the femoral head.The recombinant plasmid pCD-hVEGF165 was mixed with collagen and was implanted in the necrotic femoral head. The expression of vascular endothelial growth factor (VEGF) was examined by RNA dot hybridization and immunohistochemical techniques. Repair of the femoral head was observed by histological and histomorphometric analysis.The expression of VEGF was detected in the femoral head transfected with the VEGF gene. The femoral head transfected with the VEGF gene showed a significant increase in angiogenesis 2 and 4 weeks after gene transfection and a significant increase in bone formation 6 and 8 weeks after gene transfection on histomorphometric analysis (P < 0.01).Transfection of the VEGF gene enhances bone tissue angiogenesis. Repair of osteonecrosis could be accelerated accordingly, thus providing a potential method for therapy of osteonecrosis.
Avascular Necrosis
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This study was to find out the influence of microRNA-129-5p on proliferative ability, invasiveness, and metastasis of lung tumor cells and tumor angiogenesis. Besides, the effects of microRNA-129-5p on vascular endothelial growth factor (VEGF) level and potential regulatory mechanisms were also what we were interested in.Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay was performed to detect the microRNA-129-5p level in tumor tissues and paracancerous tissues of 50 patients with LCa, and the interaction between microRNA-129-5p expression and LCa pathological parameters was analyzed. The untreated cell group (NC) and the transfected microRNA-129-5p overexpression group (microRNA-129-5p mimics) were established, and then, the transfection efficiency of microRNA-129-5p was further verified by qRT-PCR. In H1299 and SPC-A1, cell counting kit-8 (CCK-8), Tube-formation experiments, and transwell invasion and migration tests were performed to evaluate the influence of the microRNA on the biological function of LCa cells. Finally, the potential mechanism of action of VEGF, a downstream gene of microRNA-129-5p, was explored by bioinformatics analysis and recovery experiments.QRT-PCR results showed that the level of microRNA-129-5p in cancer tissues of LCa patients was notably lower than that in normal tissues, and the difference was statistically significant. Compared with patients with highly expressed microRNA-129-5p, patients with low level had higher rates of lymph node or distant metastasis, and the overall survival rate was lower. Compared with NC group, cell proliferation, invasiveness and migration ability, and tumor angiogenesis capacity were strikingly decreased in microRNA-129-5p mimics group. Subsequently, VEGF expression was validated conspicuously enhanced in LCa cell line and tissue and was negatively correlated with microRNA-129-5p. In addition, the recovery experiment demonstrated that overexpression of VEGF could counteract the impact of microRNA-129-5p mimics on tumor angiogenesis and the invasive and migratory capacity of LCa cells, which then together led to the malignant progression of LCa.The above studies demonstrated that microRNA-129-5p was strikingly correlated with LCa lymph node or distant metastasis and poor prognosis, and it can inhibit the malignant progression of this cancer. The investigation also demonstrated that microRNA-129-5p may inhibit proliferation capacity and invasiveness of LCa cells and tumor angiogenesis via regulating VEGF.
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