期刊文献+
共找到860篇文章
< 1 2 43 >
每页显示 20 50 100
Expression of vascular endothelial growth factor and its role in oncogenesis of human gastric carcinoma 被引量:37
1
作者 Du-Hu Liu Xue-Yong Zhang Dai-Ming Fan Yu-Xin Huang Jin-Shan Zhang Wei-Quan Huang Yuan-Qiang Zhang Qing-Sheng Huang Wen-Yu Ma Yu-Bo Chai Ming Jin Institute of Digestive Disease,Xijing Hospital,~2 Department of Gastroenterology,Tangdu Hospital,~3Department of Histology and Embryology,~4 Department of Microbiology,~5 Department of Biochemistry,Fourth Military Medical University,Xi’an 710033,Shaanxi Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第4期500-505,共6页
AIM: To establish the role of vascular endothelial growth factor (VEGF) in the oncogenesis of human gastric carcinoma more directly. METHODS: The expression of VEGF and its receptor kinase-domain insert containing rec... AIM: To establish the role of vascular endothelial growth factor (VEGF) in the oncogenesis of human gastric carcinoma more directly. METHODS: The expression of VEGF and its receptor kinase-domain insert containing receptor (KDR) in human gastric cancer tissue were observed by immunohistochemical staining. VEGF levels were manipulated in human gastric cancer cell using eukaryotic expression constructs designed to express the complete VEGF(165) complimentary DNA in either the sense or antisense orientation. The biological changes of the cells were observed in which VEGF was up-regulated or down-regulated. RESULTS: VEGF-positive rate was 50%, and VEGF was mainly localized in the cytoplasm and membrane of the tumor cells, while KDR was mainly located in the membrane of vascular endothelial cells in gastric cancer tissues and peri-cancerous tissue. In 2 cases of 50 specimens, the gastric cancer cells expressed KDR, localized in both the cytoplasm and membrane. Introduction of VEGF(165) antisense into human gastric cancer cells (SGC-7901, immunofluorescence intensity, 31.6%)) resulted in a significant reduction in VEGF-specific messenger RNA and total and cell surface VEGF protein (immunofluorescence intensity, 8.9%) (P【0.05). Conversely, stable integration of VEGF(165) in the sense orientation resulted in an increase in cellular and cell surface VEGF (immunofluorescence intensity, 75.4%) (P【0.05). Lowered VEGF levels were associated with a marked decrease in the growth of nude mouse xenografted tumor (at 33 days postimplantation, tumor volume: 345.40 +/- 136.31 mm3)(P【0.05 vs control SGC-7901 group: 1534.40 +/- 362.88 mm3), whereas up-regulation of VEGF resulted in increased xenografted tumor size (at 33 days postimplantation, tumor volume: 2350.50 +/- 637.70 mm3) (P【0.05 vs control SGC-7901 group). CONCLUSION: This study provides direct evidence that VEGF plays an important role in the oncogenesis of human gastric cancer. 展开更多
关键词 gene Expression Regulation Neoplastic Adult Aged Animals Cell Division Cloning Molecular DNA Antisense DNA Complementary endothelial growth factors Endothelium vascular Female Humans LYMPHOKINES Male MICE Mice Nude Middle Aged Neovascularization Pathologic Receptor Protein-Tyrosine Kinases Receptors growth factor Receptors vascular endothelial growth factor Stomach Neoplasms Transfection Tumor Cells Cultured vascular endothelial growth factor A vascular endothelial growth factors
下载PDF
Reduction of tumorigenicity of SMMC-7721 hepatoma cells by vascular endothelial growth factor antisense gene therapy 被引量:33
2
作者 Yu Cheng Tang Yu Li Guan Xiang Qian Department of Biochemistry, Shanghai Second Medical University, Shanghai 200025, China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第1期22-27,共6页
AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cass... AIM: To test the hypothesis to block VEGF expression of SMMC-7721 hepatoma cells may inhibit tumor growth using the rat hepatoma model. METHODS: Amplify the 200 VEGF cDNA fragment and insert it into human U6 gene cassette in the reverse orientation transcribing small antisense RNA which could specifically interact with VEGF165, and VEGF121 mRNA. Construct the retroviral vector containing this antisense VEGF U6 cassette and package the replication-deficient recombinant retrovirus. SMMC-7721 cells were transduced with these virus and positive clones were selected with G418. PCR and Southern blot analysis were performed to determine if U6 cassette integrated into the genomic DNA of positive clone. Transfected tumor cells were evaluated for RNA expression by ribonuclease protection assays. The VEGF protein in the supernatant of parental tumor cells and genetically modified tumor cells was determined with ELISA. In vitro and in vivo growth properties of antisense VEGF cell clone in nude mice were analyzed. RESULTS: Restriction enzyme digestion and PCR sequencing verified that the antisense VEGF RNA retroviral vector was successfully constructed.After G418 selection, resistant SMMC-7721 cell clone was picked up. PCR and Southern blot analysis suggested that U6 cassette was integrated into the cell genomic DNA. Stable SMMC-7721 cell clone transduced with U6 antisense RNA cassette could express 200 bp small antisense VEGF RNA and secrete reduced levels of VEGF in culture condition. Production of VEGF by antisense transgene-expressing cells was 65+/-10 ng/L per 10(6) cells, 42045 ng/L per 10(6) cells in sense group and 485+/-30 ng/L per 10(6) cells in the negative control group, (P【 0.05). The antisense-VEGF cell clone appeared phenotypically indistinguishable from SMMC-7721 cells and SMMC-7721 cells transfected sense VEGF. The growth rate of the antisense-VEGF cell clone was the same as the control cells. When S.C. was implanted into nude mice, growth of antisense-VEGF cell lines was greatly inhibited compared with control cells. CONCLUSION: Expression of antisense VEGF RNA in SMMC-7721 cells could decrease the tumorigenicity, and antisense-VEGF gene therapy may be an adjuvant treatment for hepatoma. 展开更多
关键词 gene Therapy Animals Carcinoma Hepatocellular Cell Division DNA Polymerase III endothelial growth factors Endothelium vascular Enzyme-Linked Immunosorbent Assay gene Expression Humans Liver Neoplasms LYMPHOKINES MICE Mice Nude Neovascularization Pathologic Promoter Regions (genetics) RNA Antisense Research Support Non-U.S. Gov't Transduction genetic Tumor Cells Cultured vascular endothelial growth factor A vascular endothelial growth factors
下载PDF
Phosphatase and tensin homology deleted in chromosome 10,hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression
3
作者 钱群 《外科研究与新技术》 2005年第3期165-166,共2页
To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein express... To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein expression in the patients with human colorectal adenomas and adenocarcinomas.Methods The expression of PTEN,HIF-1 alpha gene was detected by using in situ hybridization,and the VEGF expression levels by immunohistochemistry in colorectal adenomas and primary colorectal adenocarcinoma.Results Strong expression of HIF-1 alpha was detectable in the majority of colorectal dadenocarcinoma,particularly surrounding areas of necrosis in adenocarcinoma.PTEN,HIF-1 alpha mRNA and VEGF protein were positive in 51.6%,67.7% and 59.7% respectively in 62 cases of adenocarcinomas,and 77.8%,44.4% and 33.3% respectively in 18 cases of adenomas.The positive rate of VEGF was higher in the patients with colorectal adenocarcinomas than that in those with adenomas,whereas that of PTEN mRNA was contrary.HIF-1 mRNA expression was correlated significantly with lymph node metastasis,liver metastasis,Duke’s stage and recurrence.During colorectal tumor progression,the expression of HIF-1 alpha mRNA was positively correlated with the VEGF protein expression (χ2= 4.751 ,P<0.05),but negatively with the PTEN mRNA expression(χ2=21.84,P<0.01).Conclusion The absence or low expression of PTEN and the increased levels of HIF-1α and VEGF may paly an important role in carcinogenesis and progression of colorectal carcinoma.These results suggest that VEGF upregulated by HIF-1 alpha gene may be involved in angiogenesis of colorectal adenocarcinoma.4 refs,1 tab. 展开更多
关键词 Phosphatase and tensin homology deleted in chromosome 10 hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression
下载PDF
Expression of Vascular Endothelial Growth Factor in Bone Morphogenetic Protein-2 Induced Osteogenesis 被引量:5
4
作者 傅德皓 《Journal of Wuhan University of Technology(Materials Science)》 SCIE EI CAS 2005年第B12期118-122,共5页
An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in bone morphogenetic protein- 2 induced osteo... An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in bone morphogenetic protein- 2 induced osteogenesis . The experimental results demonstrated that the expression signals of VEGF mRNA and VEGF appeared in cytoplasm during condensation of mesenehymal cell. As the mesenchymal cells differentiated into precartilage, the expression signals decreased in mesenehymal cells, but increased in chondrocytes and kept getting denser in the process of cartilage maturity. The peak expression of VEGF mRNA and VEGF in the experimental group appeared on the 14 th day, accompanied by numerons hypertrophic chondrocytes. When mature cartilage calcified and neu, bone trabecula formed, the expression of VEGF mRNA and VEGF decreased in chondrocytes, but still expressed moderately in the osteoblasts and osteocytes. Signals of VEGF mRNA and VEGF can not be detected in the control groups. 展开更多
关键词 bone morphogenetic protein vascular endothelial growth factor gene expression in situ hybridization IMMUNOHISTOCHEMISTRY
下载PDF
Experimental Study of Vascular Endothelial Growth Factor Gene Therapy for Avascular Necrosis of the Femoral Head 被引量:6
5
作者 杨操 杨述华 +3 位作者 杜靖远 李进 许伟华 熊宇芳 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2003年第3期297-299,316,共4页
To explore a new method for the therapy of the avascular necrosis of the femoral head, the recombinant plasmid pCD-hVEGF 165 was mixed with collagen and was implanted in the necrotic femoral head The expression... To explore a new method for the therapy of the avascular necrosis of the femoral head, the recombinant plasmid pCD-hVEGF 165 was mixed with collagen and was implanted in the necrotic femoral head The expression of vascular endothelial growth factor (VEGF) was detected by RNA dot hybridization and immunohistochemical method The repair of the femoral head was observed by histological method The results showed that the expression of VEGF was detectable in the femoral head treated with VEGF gene Angiogenesis in these femoral heads was more abundant than the control Bone repairing was augmented in the femoral head treated with VEGF gene The results suggest that angiogenesis in bone tissue could be augmented by gene transfection of VEGF and bone repairing would be accelerated accordingly 展开更多
关键词 vascular endothelial growth factor gene therapy avascular necrosis of the femoral head
下载PDF
Effect of Antisense Oligodeoxynucleotide of Vascular Endothelial Growth Factor C on Lymphangiogenesis and Angiogenesis of Pancreatic Cancer 被引量:3
6
作者 李凯 陶京 +5 位作者 李弢 许州 杨智勇 吴河水 熊炯炘 王春友 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2007年第1期51-53,共3页
In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular endothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense olig... In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular endothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografied human pancreatic cancer cells (Panc-1) was established. Thirty nude mice were randomly divided into 3 groups: PBS control group (group A), scramble-sense control group (group B) and antisense group (group C). All nude mice were treated once every 2 days as 3 times per week, for 3 weeks (oligonucleotide 10 mg/kg every time). After treatments were completed, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohistochemical method to examine microvessel density (MVD), lymphtic vessel density (LVD) of pancreatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5±41.5, 221.5±52.3 and 108.6±14.9 pg/mL in groups A, B and C respectively (P〈0.01). LVD in groups A, B and C was 13.8±2.1, 12.4±1.9 and 4.2±1.6 respectively (P〈0.01). MVD in groups A, B and C was 27.5±8.7, 25.9±4.2 and 19.4±5.6 respectively with no significant difference among the groups (P〉0.05). It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis. 展开更多
关键词 pancreatic cancer vascular endothelial growth factor C LYMPHANGIOgeneSIS ANGIOgeneSIS .gene therapy
下载PDF
Intrastriatal Gene Transfer of Vascular Endothelial Growth Factor Rescues Dopaminergic Neurons in a Rat Parkinson's Disease Model 被引量:2
7
作者 田有勇 孙圣刚 +3 位作者 汤翠菊 王家宁 陈小武 乔娴 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2006年第6期670-673,共4页
To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructe... To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructed recombinant replication-deficent adenoviral vectors carrying the gene of VEGF165 (Ad-VEGF), and injected Ad-VEGF (or Ad-LacZ and PBS as controls) into the striatum of rats 7 days after the lesion by 6-hydroxydopamine. The rat rotational behavior analysis and tyrosine hydroxylase (TH) immunohistochemistry were performed to assess the change of dopaminergic neurons. Our results showed that the rats receiving Ad-VEGF injection displayed a significant improvement in apomorphine-induced rotational behavior and a significant preservation of TH-positive neurons and fibers compared with control animals, It is concluded that intrastriatal gene transfer by Ad-VEGF may rescue the dopaminergic neurons from degeneration in a rat model of PD. 展开更多
关键词 vascular endothelial growth factor A Parkinson's disease gene therapy gene transfer technique
下载PDF
Association of vascular endothelial growth factor-634G/C and receptor for advanced glycation end products G82S gene polymorphisms with diabetic retinopathy 被引量:2
8
作者 Asmaa Kamal Khaled Abu Eleinen Ibrahem Siam 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2016年第8期1106-1111,共6页
AIMTo investigate the association of receptor for advanced glycation end products (RAGE) G82S and vascular endothelial growth factor (VEGF) -634 G/C gene polymorphisms with diabetic retinopathy (DR).METHODSOur cross-s... AIMTo investigate the association of receptor for advanced glycation end products (RAGE) G82S and vascular endothelial growth factor (VEGF) -634 G/C gene polymorphisms with diabetic retinopathy (DR).METHODSOur cross-sectional study included 61 diabetic patients, 12 of them had proliferative diabetic retinopathy (PDR), 15 had non proliferative diabetic retinopathy (NPDR), 34 had no diabetic retinopathy (NDR) and 61 healthy controls. Participants were tested for RAGE G82S and VEGF -634 G/C polymorphisms by polymerase chain reaction-restriction fragment length polymorphism.RESULTSWe found a significant association between VEGF -634 G/C polymorphism and PDR as PDR patients had increased incidence of VEGF -634 CC genotype compared to NDR patients [odds ratio for CC vs (GC+GG)=6.5, 95% CI=1.5-27.8, P=0.021]. Also VEGF -634 CC genotype and C allele were significantly higher in the PDR than in NPDR patients, which is a novel finding in our study (P=0.024, 0.009 respectively). The mean triglycerides level was significantly higher in diabetic patients with CC genotype (P=0.01) as compared to patients with other genotypes. All cases and control subjects were of the same heterozygous RAGE 82G/S genotype.CONCLUSIONPatients carrying VEGF -634 C polymorphism have a higher risk of PDR development, so VEGF -634 G/C polymorphism could be used as a predictive marker for PDR in diabetic patients. We could not find a significant association between RAGE G82S polymorphism and DR. 展开更多
关键词 diabetic retinopathy vascular endothelial growth factor receptor for advanced glycation end products gene polymorphism
下载PDF
Lentiviral-mediated vascular endothelial growth factor 165 gene transfer into neural stem cells promotes proliferation 被引量:1
9
作者 Shanshan Zhang Xiangrong Zheng Fei Yin Jielu Tan Yujia Yang 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第19期1457-1461,共5页
We constructed a lentiviral vector carrying vascular endothelial growth factor 165, which was used to transfect neural stem cells. The transfection rate was approximately 50%, as determined by flow cytometry. Vascular... We constructed a lentiviral vector carrying vascular endothelial growth factor 165, which was used to transfect neural stem cells. The transfection rate was approximately 50%, as determined by flow cytometry. Vascular endothelial growth factor protein was detected in neural stem cells and promoted proliferation. 展开更多
关键词 vascular endothelial growth factor 165 gene therapy LENTIVIRUS neural stem cells TRANSFECTION
下载PDF
Construction of human eukaryotic expression plasmid vascular endothelial growth factor 165 and its expression in transfected vascular smooth muscles 被引量:5
10
作者 Zhong-Jun Wu, Xiao-Hong Yang, Shu-Sen Zheng, Su-Fen Yang and De Shi Organ Transplant Center, First Affiliated Hospital,Zhejiang University School of Medicine, Hangzhou 310003, China Department of General Surgery, Affiliated Hospital of ZunyiMedical College, Zunyi 563003 , China and Department ofVascular Surgery, Chongqing Medical University, Chongqing 400016 , Chi-na 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2004年第3期355-359,共5页
BACKGROUND: The highly specific vascular endothelialgrowth factor (VEGF) induces the growth of vascular en-dothelial cell. This study was to construct the eukaryoticexpression plasmid of vascular endothelial growth fa... BACKGROUND: The highly specific vascular endothelialgrowth factor (VEGF) induces the growth of vascular en-dothelial cell. This study was to construct the eukaryoticexpression plasmid of vascular endothelial growth factorl65(VEGF165) and observe its expression in vascular smoothmuscles (VSMCs).METHODS: The primers were designed and synthesizedaccording to the gene sequences of human VEGF165. TheVEGF165 gene was obtained from umbilic artery tissue bythe method of RT-PCR, then it was cloned to eukaryoticexpression plasmid pBudCE4.1 by recombination strategy.The eukaryotic expression plasmid named pBudCE4.1/VEGF165 was identified by restriction enzyme digestion,and was sequenced. The pBudCE4.1/VEGF165 was trans-fected into VSMCs by using lipofection. The VEGF165 ex-pression of mRNA and protein was detected by RT-PCRand Western blot respectively.RESULTS: VEGF165 was shown about 576bp by RT-PCR.Sequencing revealed the amplified VEGF165 gene was iden-tical with that in the GeneBank. Restrictive enzyme (HindBam HI) digestion analysis showed that recombinantexpression plasmid pBudCE4. l/tVEGF165 had been con-structed successfully. The expression of VEGF165 at mRNAand protein levels in the transformed VSMCs had beendemonstrated by RT-PCR and Western blot.CONCLUSIONS: The recombinant eukaryotic expressionplasmid pBudCE4.1/VEGF165 has been successfully con-structed and expressed in transformed VSMCs. The presentstudy has laid a foundation for VEGF165 gene therapy ofvascular stenosis in the transplant organ. 展开更多
关键词 eukaryotic expression plasmid human vascular endothelial growth factor vascular smooth muscle cell gene transfer organ transplant
下载PDF
Elevation of vascular endothelial growth factor production and its effect on revascularization and function of graft islets in diabetic rats 被引量:6
11
作者 Ying Cheng Yong-Feng Liu Jia-Lin Zhang Tie-Min Li Ning Zhao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第20期2862-2866,共5页
AIM: To determine whether the elevated vascular endothelial growth factor (VEGF) expression produced by the transfected vascular endothelial cells (VECs) could stimulate angiogenesis of the graft islets and exert its ... AIM: To determine whether the elevated vascular endothelial growth factor (VEGF) expression produced by the transfected vascular endothelial cells (VECs) could stimulate angiogenesis of the graft islets and exert its effect on the graft function. METHODS: Thirty diabetic recipient rats were divided into three groups (n = 10 per group). In the control group,300 IEQ islets were transplanted in each rat under the capsule of the right kidney,which were considered as marginal grafts. In the VEC group,VEC together with the islets were transplanted in each rat. In the VEGF group,VEC transfected by pIRES2-EGFP/ VEGF165 plasmid and the islets were transplanted in each rat. Blood glucose and insulin levels were evaluated every other day after operation. Intravenous glucose tolerance test (IVGTT) was performed 10 d after the transplantation. Hematoxylin and eosin (HE) staining was used to evaluate the histological features of the graft islets. Immunohistochemical staining was used to detect insulin-6,VEGF and CD34 (MVD) expression in the graft islets. RESULTS: Blood glucose and insulin levels in the VEGF group restored to normal 3 d after transplantation. In contrast,diabetic rats receiving the same islets with or without normal VECs displayed moderate hyperglycemia and insulin,without a significant difference between these two groups. IVGTT showed that both the amplitude of blood glucose induction and the kinetics of blood glucose in the VEGF group restored to normal after transplantation. H&E and immunohistochemical staining showed the presence of a large amount of graft islets under the capsule of the kidney,which were positively stained with insulin-6 and VEGF antibodies in the VEGF group. In the cell masses,CD34-stained VECs were observed. The similar masses were also seen in the other two groups,but with a fewer positive cells stained with insulin-6 and CD34 antibodies. No VEGF-positive cells appeared in these groups. Microvessel density (MVD) was significantly higher in the VEGF group compared to the other two groups. CONCLUSION: Elevated VEGF production by trans-fected vascular endothelial cells in the site of islet transplantation stimulates angiogenesis of the islet grafts. The accelerated islet revascularization in early stage could improve the outcome of islet transplantation,and enhance the graft survival. 展开更多
关键词 Islet transplantation Revascularization vascular endothelial growth factor gene transfer vascular endothelial cells
下载PDF
Expression of Vascular Endothelial Growth Factor (VEGF) in Human Osteosarcoma Cells Transfected with Adeno-associated Virus-antisense VEGF
12
作者 徐卫国 陈安民 +1 位作者 张衣北 易成腊 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2004年第3期279-280,283,共3页
Summary: The expression of protein vascular endothelial growth factor (VEGF) in osteosarcoma cells transfected with adeno-associated virus (rAAV)-antisense VEGF was studied to provide the foundation of osteosarcoma tr... Summary: The expression of protein vascular endothelial growth factor (VEGF) in osteosarcoma cells transfected with adeno-associated virus (rAAV)-antisense VEGF was studied to provide the foundation of osteosarcoma treatment through antivascularization. The rAAV-antisense VEGF at different doses (0, 20, 50, 100, 200, 240 μl) was transfected into osteosarcoma MG-63 cell. The cells and culture supernatants were collected before and after tansfection. The expression of VEGF protein was detected by using immunohistochemical staining (SP) and Western blot. SP and Western-blot tests revealed that the MG-63 Cells transfected with rAAV-antisense VEGF had less staining than those without transfection with rAAV-antisense VEGF, and the staining intensity was negatively correlated with the doses of genes. The corresponding A values of transfected genes with different doses of rAAV-antisense VEGF (0, 20, 50, 100, 200, 240 μl) were 86 614±13 776, 73 245±15 414, 61 078±12 124, 54 657±10 953, 39 802±11 308, 32 014±15 057 respectively, with the difference being significant (P<0.05). It was concluded that the expression of VEGF protein in MG-63 cells could be inhibited by rAAV-antisense VEGF. 展开更多
关键词 adeno-associated virus antisense vascular endothelial growth factor gene transfection OSTEOSARCOMA
下载PDF
Expression of thymidine kinase mediated by a novel non-viral delivery system under the control of vascular endothelial growth factor receptor 2 promoter selectively kills human umbilical vein endothelial cells 被引量:9
13
作者 Ying Wang Hui-Xiong Xu +1 位作者 Ming-De Lu Qing Tang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第2期224-230,共7页
AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endo... AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels. 展开更多
关键词 MICROBUBBLE ULTRASOUND gene therapy vascular endothelial growth factor receptor 2 Humanumbilical vein endothelial cells
下载PDF
Inhibitory effect of antisense vascular endothelial growth factor RNA on the profile of hepatocellular carcinoma cell line in vitro and in vivo 被引量:7
14
作者 Ji-Hui Hao Ming Yu +3 位作者 Hui-Kai Li Yu-Rong Shi Qiang Li Xi-Shan Hao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第7期1140-1143,共4页
AIM: To evaluate the effect of antisense vascular endothelial growth factor (VEGF) RNA (PCMV-FGEV) transfection on the profile of hepatocellular carcinoma (HCC) SMMC-7721 cells in vitro and in vivo.METHODS: SM... AIM: To evaluate the effect of antisense vascular endothelial growth factor (VEGF) RNA (PCMV-FGEV) transfection on the profile of hepatocellular carcinoma (HCC) SMMC-7721 cells in vitro and in vivo.METHODS: SMMC-7721 cells were transfected with PCMV-FGEV antisense, PCMV-VEGF sense and empty vector plasmid encapsulated by lipofectamine as antisense group, sense group and control group respectively. The positive cell clones were selected with G418. The stable transfection and expression of VEGF in the cells were determined by RT-PCR and immunohistochemistry. Cell proliferation was observed by Mnassay. FACS analysis was used to determine the effect of PCMV-FGEV transfection on cell apoptosis. The growth of transfected cells in vivo was also observed in nude mice.RESULTS: VEGF expression was reduced in SMMC-7721 transfected with PCMV-FGEV, which was confirmed by RT-PCR and immunohistochemistry. No effect of PCMV- FGEV transfection was found on cell proliferation and cell apoptosis of SMMC-7721 in vitro. The growth of cells transfected with PCMV-FGEV was slow in nude mice and accompanied with obvious apoptosis. The latent time of tumors in the antisense group was 25.0 :l: 1.8 d, which was longer than that in sense and control groups (F= 19.455, P〈 0.01). The average tumor weight in antisense group (0.96 g±0.28 g) was the smallest among the three groups (F= 21.501, P〈 0.01).CONCLUSION: The expression of VEGF can be inhibited by antisense PCMV-FGEV. Antisense PCMV-FGEV has no effect on cell proliferation and apoptosis of SMMC-7721 in vitro but can inhibit tumor growth and induce cell apoptosis in vivo. 展开更多
关键词 Antisense RNA vascular endothelial growth factor gene expression Hepatocellular carcinoma TRANSFECTION
下载PDF
Abnormal expression of vascular endothelial growth factor (VEGF) and its clinical features in tissues of human lung cancer 被引量:3
15
作者 Xinhua Wu Dengfu Yao +4 位作者 Gongshen Shi Liwei Qiu Wei Wu Songshi Ni Xueguang Zhang 《Journal of Nanjing Medical University》 2005年第4期194-198,共5页
Objective: Angiogennesis, the formation of new blood vessels from the existing vascular bed, is essential step for growth and invasion of primary rumor. Vascular endothelial growth factor (VEGF) is known to play cr... Objective: Angiogennesis, the formation of new blood vessels from the existing vascular bed, is essential step for growth and invasion of primary rumor. Vascular endothelial growth factor (VEGF) is known to play crucial role in tumor angiogenesis. In the present study, we investigate the expression of VEGF and VEGF-mRNA in the angiogennesis, metastasis and prognosis of lung cancer. Methods: The VEGF cellular distributions and expression in 38 specimens of patients with lung cancer were investigated with immunohistochemistry stain technology. The total RNAs in 38 tissues of lung cancer was measured, then the levels of VEGF-mRNA expression were analyzed by a reverse-transcription polymerase chain reaction (RT-PCR) assay. The levels of VEGF in sera of patients with lung cancer, benign lung diseases and healthy controls were detected through Enzyme linked immunosorbent assay (ELISA) method. Results: The VEGF positive stain was 76% in 38 cases of lung cancer specimens. The 89% rate of VEGF stain was found for clinical stage Ⅲ cases and 92% for stage Ⅳ lung cancers. The significantly higher expression of VEGF was evidenced in patients with lymph node metastasis (84%), distant metastasis (90%), and lung cancers with lower histological differentiation (89%), respectively. The expression level of total RNA was significantly higher in patients with lung cancers than that in their paracancerons or distant lung tissues. The VEGF expressions were tightly correlated with total RNA concentration of lung carcinoma ( P 〈 0.01 ). The predominant expressions of VEGF121 and VEGF165 gene fragments were found in lung cancer specimens by RT-PCR analysis. No significant difference of serum VEGF levels was detected between cases with lung cancer and patients with benign diseases. However, the VEGF level of cases with benign diseases was decreased significantly after patients with anti-inflammation medication. Conclusion: The present data suggested that the rumor tissue VEGF expression and VEGF-mRNA analysis in patients with lung cancer be a useful indicator for angiogenesis and metastasis of lung cancer. 展开更多
关键词 vascular endothelial growth factor lung cancer gene amplification immuno-histochemlstry ELISA
下载PDF
The Mechanical Study of Vascular Endothelial Growth Factor on the Prevention of Restenosis after Angioplasty
16
作者 刘启功 陆再英 +2 位作者 周洪莲 颜进 张卫东 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2001年第3期195-197,共3页
The mechanism of vascular endothelial growth factor (VEGF) on the prevention of restenosis after angioplasty was investigated. The cultured vascular endothelial cells (VEC) were incubated with the conditioned medium (... The mechanism of vascular endothelial growth factor (VEGF) on the prevention of restenosis after angioplasty was investigated. The cultured vascular endothelial cells (VEC) were incubated with the conditioned medium (CM) from vascular smooth muscle cells (VSMC) infected with recombinant adenoviruses containing the hVEGF 165 gene. To observe the effects of VEGF on proliferation and NO, ET, 6-keto-PGF1α secretion of VEC, WST-1 method, Griess method and radioimmunoassay were used respectively. The PDGF-B mRNA transcription in VECs was detected by RT-PCR. It was showed that NO, 6-keto-PGF1α and OD value were markedly increased in a dose-dependent manner in the VEGF-treated groups as compared with those in the control group, while ET and PDGF-B mRNA were significantly decreased in the VEGF-treated groups (P<0.05 or P<0.01). Adenovirus vector mediated hVEGF 165 gene could promote the proliferation of VECs and improve NO, PGI 2 secretion, inhibit ET secretion and PDGF-B mRNA transcription in the VECs. The above results offered further theoretical evidence for VEGF on the prevention of restenosis after angioplasty. 展开更多
关键词 vascular endothelial growth factor ADENOVIRUS gene therapy vascular endothelial cell RESTENOSIS
下载PDF
Human Vascular Endothelial Growth Factor cDNA Cloning and Expression in Osteoblasts
17
作者 杨操 杨述华 屈伸 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2001年第2期134-137,共4页
Human vascular endothelial growth factor cDNA was amplified by nested polymerase chain reaction method from the HL60 cells. Then a pCD hVEGF 165 recombinant plasmid was constructed. Rabbit osteoblasts were tra... Human vascular endothelial growth factor cDNA was amplified by nested polymerase chain reaction method from the HL60 cells. Then a pCD hVEGF 165 recombinant plasmid was constructed. Rabbit osteoblasts were transfected with pCD hVEGF 165 plasmid by lipofectin mediated gene transfer. The transient expressive results were detected by immunohistochemical method. It was observed that the expression of human VEGF gene was detected 72 h after transfecting distinctly. 展开更多
关键词 vascular endothelial growth factor gene amplification gene expression
下载PDF
Vascular endothelial growth factor,p53,and the H-ras oncogene in Egyptian patients with bladder cancer
18
作者 Farha A El-Chennawi Fatma A Auf +5 位作者 Shereen S Metwally Youssef M Mosaad Atallah A Shaaban Mahmoud Abdo El-Baz Ziyad E Tawhid Zakaria F Lotfy 《World Journal of Gastrointestinal Oncology》 SCIE CAS 2009年第1期62-68,共7页
AIM:To evaluate the relationship between vascular endothelial growth factor(VEGF),p53,and the H-ras oncogene and different clinicopathological parameters in Egyptian patients with Schistosoma-associated transitional c... AIM:To evaluate the relationship between vascular endothelial growth factor(VEGF),p53,and the H-ras oncogene and different clinicopathological parameters in Egyptian patients with Schistosoma-associated transitional cell carcinoma of the bladder.METHODS:The study included 50 patients with transitional cell carcinoma for whom radical cystectomy and urinary diversions were carried out.VEGF and p53 protein expressions were evaluated with an immunohistochemical staining method,and H-ras oncogene mutations were analyzed with a polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) technique.RESULTS:High grade tumors revealed higher p53 immunostaining than low grade tumors(P = 0.016).p53 and VEGF protein expressions,as well as H-ras oncogene mutations,had an insignificant impact on patient outcomes(P = 0.962,P = 0.791,and P = 967,respectively).Cancer extension to regional lymph nodes was associated with poor outcomes(P = 0.008).CONCLUSION:VEGF,p53 and the H-ras oncogene have no relation to patient survival and outcome in Schistosoma-associated transitional cell carcinoma. 展开更多
关键词 Bladder cancer Transitional cell carcinoma vascular endothelial growth factor p53 H-RAS ONCOgene
下载PDF
Efficacy and safety of therapeutic angiogenesis from direct myocardial administration of an adenoviral vector expressing vascular endothelial growth factor 165
19
作者 张端珍 盖鲁粤 +2 位作者 范瑞云 董蔚 文应峰 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第5期643-648,145,共6页
OBJECTIVE: To investigate whether direct administration of adenoviral vectors (Ad) containing the complementary deoxyribonucleic acid (cDNA) of vascular endothelial growth factor 165 (Ad-VEGF165) induces porcine coron... OBJECTIVE: To investigate whether direct administration of adenoviral vectors (Ad) containing the complementary deoxyribonucleic acid (cDNA) of vascular endothelial growth factor 165 (Ad-VEGF165) induces porcine coronary collateral vessel formation, improves regional myocardial perfusion and function and is safe. METHODS: Three weeks after miniature swine underwent left thoracotomy and placement of an Ameroid constrictor on the left circumflex coronary artery (LCX), Ad-VEGF165 (n = 6) or the control, Ad expressing beta-galactosidase cDNA (Ad-Gal, n = 6), was directly administered into the ischemic myocardium in the circumflex distribution. All animals were sacrificed 4 wk after the second surgery. Myocardial perfusion and function were assessed by electrocardiogram-gated single photon emission computed tomography (GSPECT) imaging. Ex vivo coronary angiography was performed to examine collateral vessels. Toxicity was assessed by blood analyses on the day just before (day 0) and on day 1, 3, 7, 28 after vector delivery and by vascular, myocardial and liver histology after sacrifice. RESULTS: GSPECT imaging 4 wk after administration of Ad-VEGF165 demonstrated significant reduction in ischemic area (P 展开更多
关键词 ADENOVIRIDAE Animals Collateral Circulation Coronary Angiography Coronary Vessels DNA Complementary Electrocardiography endothelial growth factors Female gene Therapy gene Transfer Techniques genetic Vectors LYMPHOKINES Male Myocardial Ischemia Neovascularization Physiologic SWINE Swine Miniature Tomography Emission-Computed Single-Photon Treatment Outcome vascular endothelial growth factor A vascular endothelial growth factors
原文传递
Effect of vascular endothelial growth factor 165 gene transfection on bone defects and its mRNA expression in rabbits 被引量:11
20
作者 ZHAO Dong-mei WANG Hai-bin +5 位作者 YANG Jia-feng WU Shi-qing LIU Jun-li XU Fu-yu QIU Li-ping CAI Jing-long 《Chinese Medical Journal》 SCIE CAS CSCD 2007年第13期1187-1191,共5页
Background Gene therapy has been a hot spot in repair of bone defects in recent years. This study aimed to construct a recombinant plasmid pcDNA3.1-VEGF165, and to observe the effect of vascular endothelial growth fac... Background Gene therapy has been a hot spot in repair of bone defects in recent years. This study aimed to construct a recombinant plasmid pcDNA3.1-VEGF165, and to observe the effect of vascular endothelial growth factor 165 (VEGF165) gene therapy on bone defects in rabbits. Methods Total RNA was extracted from rabbit bone tissues. VEGF165 cDNA fragment was prepared by reverse transcription and the gene was cloned by polymerase chain reaction (PCR). Plasmid pMD18-T/VEGF165 combined with pcDNA3.1 was cloned to reconstruct pcDNA3.1-VEGF165 plasmid. Thirty New Zealand white rabbits weighing (2.50±0.13)kg were used to establish models of bone defects (1 cm in length) of the bilateral radii. The bone defects were repaired with absorbable gelatin sponge. After the operation, physiological sodium chloride solution was injected into the injured site in one of the forelegs of the rabbits as the control group, and pcDNA3.1-VEGF165 plasmid (0.2 ml, 200 ng) was injected into the opposite foreleg as the experiment groups. At weeks 1, 2, 4, 6, 8, and 12 after the treatments, the bones were examined by X-ray, and the specimens of the bone defects were collected, stained with HE, and observed under a light microscope. The expression of VEGF165 mRNA was examined by real-time quantitative polymerase chain reaction (RQ-PCR). Results The pcDNA3.1-VEGF165 plasmid with a correct sequence was constructed successfully. Postoperative X-ray found no difference between the two groups at week 1. In the experiment group, callus and synostosis were observed after 2 weeks, and osteosis structure was normal at week 12; these phenomena occurred much later in the control group. In the experiment group, HE staining showed a large amount of newly formed blood vessels after 2 weeks, a number of bone trabeculae with osteoblasts proliferation at 4 weeks, and fresh bone cortex and reformed medullary cavity at 12 weeks; whereas in the control group these structures formed in later phases. The VEGF165 mRNA in the experiment group was expressed at a low level at week 1, reached the peak at weeks 3, and then decreased to a normal level after 6 weeks. Conclusions Local use of pcDNA3.1-VEGF165 plasmid at bone defects can upregulate the expression of VEGF165 and accelerate the formation of capillaries and the repair of bone defects. Angiogenesis and osteogenesis can be promoted by a combination of pcDNA3.1-VEGF165 and gelatin sponge. 展开更多
关键词 gene transfection vascular endothelial growth factor bone defects
原文传递
上一页 1 2 43 下一页 到第
使用帮助 返回顶部