Abnormal expression of vascular endothelial growth factor (VEGF) and its clinical features in tissues of human lung cancer

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.

Nerve growth factor alleviates cerebral infarction and neurologic deficits by regulating VEGF,SDF-1 and S100A12 expression through PI3K pathway

Stroke remains the leading cause of death and disability worldwide,which destroys the quality of patients’lives and thus is becoming a heavy burden to the society.However,the current therapeutic approaches are far from satisfaction.The objective of this study is to elucidate the impact of nerve growth factor(NGF)on the brain damage induced by cerebral ischemia and its potential molecular mechanism.Middle cerebral artery occlusion(MCAO)rats were used as animal models and neurological functions were evaluated by modified Neurological Severity Score(NSS).Brain cell apoptosis was analyzed by TUNEL-positive staining while brain infarct size was determined according to 2% 2,3,5-triphenyltetrazolium chloride(TCC)staining volume.Rats receiving NGF demonstrated significantly alleviated brain damage,reflected by a substantial improvement in the neurobehavioral outcome,a decrease in brain cell apoptosis and shrinkage of brain infarct volume.Further analysis revealed a markedly elevated circulating vascular endothelial growth factor(VEGF)and stromal cell-derived factor 1(SDF-1)levels as well as a significant downregulation of SA10012 expression in NGF treated group compared with the untreated group.Strikingly,the protective effect of NGF on cerebral ischemic injury was abolished in rats treated with both NGF and PI3K inhibitors,indicating that phosphoinositide-3-kinase(PI3K)signaling is essential for NGF function.In conclusion,NGF treatment might be a potential therapeutic approach against cerebral infarction by downregulating SA10012 expression and upregulating VEGF,SDF-1 in a PI3K signaling dependent manner.

Hypoxia-inducible factor 1alpha and vascular endothelial growth factor in Glioblastoma Multiforme:a systematic review going beyond pathologic implications

Glioblastoma multiforme(GBM)is an aggressive primary brain tumor characterized by extensive heterogeneity and vascular proliferation.Hypoxic conditions in the tissue microenvironment are considered a pivotal player leading tumor progression.Specifically,hypoxia is known to activate inducible factors,such as hypoxia-inducible factor 1alpha(HIF-1α),which in turn can stimulate tumor neo-angiogenesis through activation of various downward mediators,such as the vascular endothelial growth factor(VEGF).Here,we aimed to explore the role of HIF-1α/VEGF immunophenotypes alone and in combination with other prognostic markers or clinical and image analysis data,as potential biomarkers of GBM prognosis and treatment efficacy.We performed a systematic review(Medline/Embase,and Pubmed database search was completed by 16th of April 2024 by two independent teams;PRISMA 2020).We evaluated methods of immunoassays,cell viability,or animal or patient survival methods of the retrieved studies to assess unbiased data.We used inclusion criteria,such as the evaluation of GBM prognosis based on HIF-1α/VEGF expression,other biomarkers or clinical and imaging manifestations in GBM related to HIF-1α/VEGF expression,application of immunoassays for protein expression,and evaluation of the effectiveness of GBM therapeutic strategies based on HIF-1α/VEGF expression.We used exclusion criteria,such as data not reporting both HIF-1αand VEGF or prognosis.We included 50 studies investigating in total 1319 GBM human specimens,18 different cell lines or GBM-derived stem cells,and 6 different animal models,to identify the association of HIF-1α/VEGF immunophenotypes,and with other prognostic factors,clinical and macroscopic data in GBM prognosis and therapeutic approaches.We found that increased HIF-1α/VEGF expression in GBM correlates with oncogenic factors,such as miR-210-3p,Oct4,AKT,COX-2,PDGF-C,PLDO3,M2 polarization,or ALK,leading to unfavorable survival.Reduced HIF-1α/VEGF expression correlates with FIH-1,ADNP,or STAT1 upregulation,as well as with clinical manifestations,like epileptogenicity,and a favorable prognosis of GBM.Based on our data,HIF-1αor VEGF immunophenotypes may be a useful tool to clarify MRI-PET imaging data distinguishing between GBM tumor progression and pseudoprogression.Finally,HIF-1α/VEGF immunophenotypes can reflect GBM treatment efficacy,including combined first-line treatment with histone deacetylase inhibitors,thimerosal,or an active metabolite of irinotecan,as well as STAT3 inhibitors alone,and resulting in a favorable tumor prognosis and patient survival.These data were supported by a combination of variable methods used to evaluate HIF-1α/VEGF immunophenotypes.Data limitations may include the use of less sensitive detection methods in some cases.Overall,our data support HIF-1α/VEGF’s role as biomarkers of GBM prognosis and treatment efficacy.

The effect of AD-VEGF-siRNA on the expression of vascular endothelial growth factor in osteosarcoma-bearing nude mice

Objective： To explore the influence of AD-VEGF-siRNA on the expression of vascular endothelial growth factor （VEGF） in neoplasm and blood serum. Methods： Transplantable model of human osteosarcoma was successfully established by the way of subcutaneous injection of VEGF highly expressed human MG63 osteosarcoma cells. These mice were divided randomly into three groups： AD-VEGF-siRNA group, 15 mice; AD-EGFP group, 15 mice; PBS group, 15 mice. Three mice were additionally raised without any treatment. The drug was injected intratumorally 200 μL at each time, once a day. The total dose of virus was 2 × 10^9 pfu. Three osteosarcema-bearing mice of each group were sacrificed at 11th, 14th ,17th day after the implantation of MG63 cells. The expression of VEGF in implanted tumors and blood serum was detected by ELISA methods. Then the left mice were all sacrificed at the end of experiment （19th day）. The expression of VEGF in implanted tumors was detected by RT-PCR and immune histochemistry methods, and that in implanted tumors and blood serum was detected by ELISA methods. Results： （1） Tumors in mice could be seen at 5th day from the implantation of MG63 cells. （2） The expression of VEGF could be detected in all groups by RT-PCR and immune histochemistry, Which was much lower in the group receiving AD-VEGF-siRNA therapy than two control groups （P 〈 0.05）. （3） The expression of VEGF in blood serum of osteosarcoma-bearing mice was much higher than that of three healthy mice by ELISA （P 〈 0.05）. （4） The expression of VEGF in blood serum and neoplasm in AD-VEGF-siRNA group was much lower than that in two control groups （P 〈 0.05）. Conclusion： AD-VEGF-siRNA could effectively inhibited VEGF expression in vivo. This technology would bring some good references for our therapy of antiangiogenesis in osteosarcoma.

IMPACTS ON THE EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN ISCHEMIC BRAIN TISSUE TREATED WITH CLUSTER PUNC-TURE ON SCALP ACUPOINTS IN RATS

Objective To probe into the impacts on the expression of vascular endothelial growth factor （VEGF） in ischemic brain tissue treated with cluster puncture on scalp acupoints in rats, Methods 128 rats were randomized into pseudo-operation group, model group, scalp-needling group and cluster-needling group, In scalp-needling group, penetration method was used on the focal side from Bǎihuì （百会 GV20） to Qǔbīn （曲鬓 GB7）, and in cluster-needling group, penetration method was used on both sides from GV20 to GB7, and a common needling on GV 20. Needles were punctured 2 mm in depth, constantly rotated for 10min, retained for 2 h. Immunohistrochemical method was applied to determine VEGF expression and microvessel density （MVD）. Results With the intervention of cluster puncture on scalp acupoints, VEGF： expressions on every time-spot after ischemia were enhanced apparently, superior to those in scalp-needling group. On the three time-spots of the 7^th, 14^th and 21^st days, MVD was increased after cluster puncture on scale acupoints, superior to those in scalp-needling group. Conclusion Cluster puncture on scalp acupoints up-regulated VEGF expression and promoted regeneration of microvessel.

益气化瘀汤联合羟苯磺酸钙治疗糖尿病肾病气虚血瘀证的疗效及对VEGF,IGF-1表达水平的影响研究

【目的】观察益气化瘀汤(由黄芪、山药、茯苓、炒芡实、旱莲草、金樱子、焦山楂、女贞子、丹参、益母草等组成)联合羟苯磺酸钙治疗糖尿病肾病(DN)气虚血瘀证的临床疗效及对血管内皮生长因子(VEGF)、胰岛素样生长因子1(IGF-1)的影响。【方法】将90例DN气虚血瘀证患者随机分为观察组和对照组,每组各45例。所有患者均接受基础降糖治疗和控制血压、调节脂代谢紊乱等治疗。在此基础上,对照组患者给予羟苯磺酸钙治疗,观察组患者在对照组的基础上联合益气化瘀汤治疗,疗程为3个月。观察2组患者治疗前后中医证候积分、肾功能指标及血清VEGF、IGF-1水平的变化情况,并评价2组患者的临床疗效。【结果】(1)疗效方面,治疗3个月后,观察组的总有效率为91.11%(41/45),对照组为75.56%(34/45),组间比较(χ2检验),观察组的疗效明显优于对照组(P<0.05)。(2)中医证候积分方面,治疗1个月和3个月后,2组患者的中医证候积分均较治疗前明显降低(P<0.05),且治疗3个月后又均较治疗1个月后明显降低(P<0.05);组间比较,观察组在治疗1个月和3个月后对中医证候积分的降低作用均明显优于对照组(P<0.01)。(3)肾功能指标方面,治疗后,2组患者的血肌酐(Scr)、尿素氮(BUN)、肾小球滤过率(GFR)等肾功能指标均较治疗前明显改善(P<0.05),且观察组对各项肾功能指标的改善作用均明显优于对照组(P<0.01)。(4)血清VEGF、IGF-1水平方面,治疗后,2组患者的血清VEGF、IGF-1水平均较治疗前明显降低(P<0.05),且观察组对血清VEGF、IGF-1水平的降低作用均明显优于对照组(P<0.01)。(5)治疗过程中,2组患者均无明显不良反应发生,具有较高的安全性。【结论】益气化瘀汤联合羟苯磺酸钙治疗DN气虚血瘀证患者疗效确切,可有效下调血清VEGF、IGF-1水平,明显改善患者肾功能,显著减轻患者临床症状,且具有较高的安全性。

缺血性视网膜静脉阻塞继发黄斑水肿患者基线血清己糖激酶1抗体滴度与抗VEGF治疗后视力改善的相关性分析

目的分析缺血性视网膜静脉阻塞继发黄斑水肿(RVO-ME)患者基线血清己糖激酶1抗体滴度与抗血管内皮生长因子(VEGF)治疗后视力改善的相关性。方法招募2017年6月至2020年2月在首都医科大学宣武医院确诊为缺血性RVO-ME并接受初始抗VEGF治疗的53例患者,其中缺血性视网膜中央静脉阻塞(CRVO)23例(CRVO组),缺血性视网膜分支静脉阻塞(BRVO)30例(BRVO组)。另选取该院同期30例行超声乳化的白内障患者作为对照组。研究对象行基线血清己糖激酶1抗体滴度检测、眼科常规检查和光学相干断层成像(OCT)检查。所有RVO-ME患者按照“3+按需治疗方案(pro re nata,PRN)”向玻璃体内注射抗VEGF药物治疗。随访12个月,采用多元线性回归分析缺血性RVO-ME患者抗VEGF治疗后视力改善的影响因素。结果CRVO组基线logMAR BCVA高于对照组和BRVO组,CRVO组和BRVO组基线CRT、基线血清己糖激酶1抗体滴度高于对照组,且CRVO组基线CRT、基线血清己糖激酶1抗体滴度高于BRVO组,差异有统计学意义(P<0.05)。RVO-ME患者基线血清己糖激酶1抗体滴度与随访6个月(r=0.377,P=0.005)、9个月(r=0.362,P=0.008)和12个月(r=0.465,P<0.001)时BCVA改善呈正相关,与随访12个月时中断EZ横向长度减少值(r=0.401,P=0.001)呈正相关。多元线性回归分析结果显示,基线logMAR BCVA、基线血清己糖激酶1抗体滴度是缺血性RVO-ME患者抗VEGF治疗随访12个月时BCVA改善的影响因素(P<0.05)。结论己糖激酶1抗体作为一种新的血清生物标志物,与缺血性RVO-ME患者抗VEGF治疗后的视力改善相关。

罗沙司他对腹膜透析大鼠腹膜HIF-1α/VEGF信号通路表达及纤维化的影响

目的评估罗沙司他(Roxadustat,ROX)对腹膜透析(Peritoneal dialysis,PD)大鼠腹膜组织的缺氧诱导因子-1α(Hypoxia-inducible factors-1α,HIF-1α)以及血管内皮生长因子(Vascular endothelial growth factor,VEGF)表达的影响,同时探讨其对腹膜纤维化程度的潜在作用及其机制。方法购入30只雄性SPF级大鼠,采用5/6肾脏切除法成功构建尿毒症腹膜透析大鼠模型27只,随机分为空白对照组、PD组、PD+ROX组(联合给予5 mg/kg ROX,每周3次),每组9只,持续给药1个月,在灌胃后6、12、24 h以及4周时,采用ELISA法检测腹膜透析液、血液中HIF-1α和VEGF的浓度。利用HE和Masson染色观察大鼠腹膜组织病理变化,WB和免疫组化检测腹膜组织中HIF-1α、VEGF蛋白表达变化。结果HE和Masson染色观察各组大鼠腹膜组织,与空白对照组比较,PD组大鼠的腹膜组织出现了明显的纤维化和血管增生现象;与PD组相比,PD+ROX组大鼠的腹膜组织纤维化和血管增生现象明显改善。PD组腹膜组织中HIF-1α和VEGF蛋白表达水平显著高于空白对照组(P均<0.05),增加ROX干预后,PD+ROX组大鼠腹膜组织中HIF-1α和VEGF蛋白表达水平显著降低(P均<0.05)。PD+ROX组血清中HIF-1α和VEGF浓度均高于空白对照组和PD组(P均<0.05)。PD+ROX组腹膜透析液中HIF-1α和VEGF浓度均低于空白对照组和PD组(P均<0.05)。结论ROX治疗能有效降低腹膜组织及腹膜透析液中的HIF-1α和VEGF表达水平,缓解由葡萄糖腹膜透析引起的腹膜组织纤维化和血管增生。同时,血清中HIF-1α和VEGF水平的提升有助于改善腹膜组织的缺氧状况,抑制HIF-1α/VEGF信号通路,对抗腹膜纤维化。

Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma

BACKGROUND: Angiogenesis is known to be essential to the survival, growth, invasion, and metastasis of tumor cells. Vascular endothelial growth factor (VEGF) are an important angiogenic factor regulating tumor angiogenesis, but its significance and tumor pathologic features are un- clear in hepatocellular carcinoma (HCC). In the present study, we analyzed expression of tissue VEGF, alteration of microvascular density (MVD) in microvessel angiogenesis, development and metastasis of HCC, and level of serum VEGF in differential diagnosis of benign and malignant liv- er diseases. METHODS: Tumor specimens were prospectively collected from HCC patients undergoing resection. Total RNAs were extracted and the expression levels were detected from different parts of HCC tissues. The cellular distributions of VEGF and MVD of liver tumors and their paracancerous and distal cancerous tissues were investigated by streptavi- din peroxidase (S-P) immunohistochemistry, respectively. The VEGF levels of circulating blood and hepatoma tissues were measured by enzyme-linked immunosorbent assay. RESULTS: The incidence of VEGF expression was 63.9% in HCCs (23/36 cases), 78.3% in non-encapsulated HCCs (18/23), and 90.9% in HCCs with extrahepatic metastasis (10/11), respectively. The VEGF expression was tightly correlated with MVD (P <0.01). The MVD in HCC with metastasis, low differentiation or non-encapsulation was significantly higher than that in HCC with intact capsule, high differentiation, or no metastasis. No significant diffe- rence was found between VEGF, MVD, tumor size, and hepatitis virus infection. The level of total RNA in HCC tis- sues was significantly lower but the VEGF level significantly higher than those in paracancerous or distal cancerous ones (P<0.01). The abnormal expression levels of VEGF in sera of HCC patients were directly correlated with the me- tastasis and recurrence of tumors. CONCLUSION: The high expression of VEGF and abnor- mality of tissue MVD are useful predictors for vascular inva- sion and metastasis of liver tumors.

Impact of Serum Vascular Endothelial Growth Factor on Prognosis in Patients with Unresectable Hepatocellular Carcinoma after Transarterial Chemoembolization

Objective： To investigate the expression level with unresectable hepatocellular carcinoma （HCC） and to assess the impact of serum VEGF as chemoembolization （TACE） treatments. of serum vascular endothelial growth factor （VEGF） in patients and its relationship with the c a predictive factor for HCC nicopathological characteristics, prognosis during transarterial Methods： Serum VEGF levels were measured using enzyme-linked immunosorbent assay （ELISA） in 60 random patients who underwent TACE or transarterial infusion （TAI） for unresectable HCC between May and September 2008 and 12 healthy volunteers were also involved in this study to serve as control. All patients＇ clinicopathological features were retrospectively analyzed. Serum VEGF levels were correlated with clinicopathological features of the HCC patients. The patients＇ survival rates were analyzed with Kaplan-Meier survival curves and compared by the log-rank test. The prognostic significance of serum VEGF levels and factors related to survival rate were evaluated by univariate and multivariate analysis. Results： The median serum VEGF level in the HCC patients was 285 pg/ml （range 14-1,207 pg/ml）, significantly higher than that of healthy controls （P=0.021）. The serum VEGF levels were significantly correlated with platelet counts （r=0.396, P=0.002） but not other clinicopathological features. Patients with serum VEGF level 〉285 pg/ml had worse overall survival compared with those with serum VEGF level 〈285 pg/ml （P=0.002）. By multivariate analysis, the serum VEGF level was a significant prognostic factor. Conclusion： High serum VEGF levels may predict poor prognosis of HCC after TACE. This study highlights the importance of tumor biomarker as a prognostic predictor in TACE therapy for HCC, which has an intrinsic problem of unavailability of histopathological prognostic features.

Apelin and vascular endothelial growth factor are associated with mobilization of endothelial progenitor cells after acute myocardial infarction

This study was designed to determine the levels of early endothelial progenitor cells （EPCs）, apelin, vascu- lar endothelial growth factor （VEGF） and stromal cell-derived growth factor-1 （SDF-1） after acute myocardial infarction （AMI）, and to investigate the relationships between these cytokines and early EPCs. Early EPCs, de- fined as CD133＋, KDR＋, and CD34~ cells, were quantified by flow cytometry. The levels of early EPCs and those cytokines in AMI patients were significantly different from those with coronary artery disease or controls （P 〈 0.05）. Plasma apelin levels were inversely correlated with Gensini score and early EPCs （both P 〈 0.01）. Early EPCs, VEGF and SDF-1 showed different patterns of changes in AMI patients during the first 24 h. The trend in the change of early EPCs was proportionally correlated with that of VEGF （P 〈 0.05）. AMI patients exhibited in- creased early EPCs with remarkably decreased apelin levels and enhanced VEGF levels.

The Effect of Simvastatin on mRNA Expression of Transforming Growth Factor-β1,Bone Morphogenetic Protein-2 and Vascular Endothelial Growth Factor in Tooth Extraction Socket

Aim To determine the effect of local simvastatin application on the mRNA expression level of transforming growth factor-β1 （TGF-β1）, bone morphogenetic protein-2 （BMP-2） and vascular endothelial growth factor （VEGF） in the tooth sockets of rat. Methodology Forty-eight male Wistar rats were randomly divided into experimental and control groups （n=24）. Polylactic acid/polyglycolic acid copolymer carriers, with or without simvastatin, were implanted into extraction sockets of right mandibular incisors. The expression of TGF-β1, BMP-2 and VEGF mRNA was determined by in situ hybridization in the tooth extraction socket at five days, one week, two weeks and four weeks after implantation. Results The fusiform stroma cells in the tooth extraction socket began to express TGF-β1, BMP-2 and VEGF mRNA in both experimental and control groups from one week after tooth extraction until the end of experiment. The expression of TGF-131 and BMP-2 mRNA in the experimental group was significantly up-regulated after one, two and four weeks, and expression of VEGF mRNA was significantly increased after one and two weeks compared with that in the control group. Conclusion The findings indicate that local administration of simvastatin can influence alveolar bone remodeling by regulating the expression of a school of growth factors which are crucial to osteogenesis in the tooth extraction socket.

Serum Vascular Endothelial Growth Factor Levels in Patients with Non-small Cell Lung Cancer and Its Relations to the Micrometastasis in Peripheral Blood

To examine the relationship between the levels of the serum vascular endothelial growth factor （VEGF） and the micrometastasis of peripheral blood in patients with non-small cell lung cancer （NSCLC）, 108 NSCLC patients, including 40 patients with benign lung diseases and 30 healthy controls, were investigated. The serum VEGF levels were detected by ELISA and CK19 mRNA in peripheral blood by reverse transcriptase-polymerase chain reaction （RT-PCR）. In NSCLC group, the serum VEGF levels and the positive rate of CK19 mRNA in peripheral blood were 479.8±268.5 pg/mL and 66.7%, which were significantly higher than those of the other two groups respectively （P〈0.01）, and both of them were increased significantly with the progression of clinical stage of the tumors （P〈0.01）. Serum VEGF levels as well as the positive rate of CK19 mRNA in different pathological types of lung cancer had no significant differences （P〉0.05）. Serum VEGF levels in the patients positive for CK19 mRNA was 561.7±325.6 pg/mL. It is significantly higher than that in the negative patients （P〈0.01）. There existed a significant correlation between serum VEGF levels and expression of CK19 mRNA in peripheral blood in NSCLC patients （P〈0.001）. The detection of serum VEGF levels and CK19 mRNA in peripheral blood is helpful in judging the condition and the prognosis of NSCLC patients, and serum VEGF levels and CK19 mRNA are independent of the pathological types of lung cancer. The micrometastasis in peripheral blood of NSCLC patients is significantly associated with serum VEGF levels.

STI571 (Glivec) suppresses the expression of vascular endothelial growth factor in the gastrointestinal stromal tumor cell line,GIST-T1

AIM： To estimate whether S-TI571 inhibits the expression of vascular endothelial growth factor （VEGF） in the gastrointestinal stromal tumor （GIST） cells. METHODS： We used GIST cell line, GIST-T1. It has a heterogenic 57-bp deletion in exon 11 to produce a mutated c-KIT, which results in constitutive activation of c-KIT. Cells were treated with/without STI571 or stem cell factor （SCF）. Transcription and expression of VEGF were determined by RT-PCR and flow cytometry or Western blotting, respectively. Activated c-KIT was estimated by immunoprecipitation analysis. Cell viability was determined by PITT assay. RESULTS： Activation of c-KIT was inhibited by STI571 treatment. VEGF was suppressed at both the transcriptional and translational levels in a temporal and dose-dependent manner by STI571. SCF upregulated the expression of VEGF and it was inhibited by S-13571. STI571 also reduced the cell viability of the GIST-T1 cells, as determined by PTT assay. CONCLUSION： Activation of c-KIT in the GIST-T1 regulated the expression of VEGF and it was inhibited by ST571. STI571 has antitumor effects on the GIST cells with respect to not only the inhibition of cell growth, but also the suppression of VEGF expression.

A fusion protein containing murine vascular endothelial growth factor and tissue factor induces thrombogenesis and suppression of tumor growth in a colon carcinoma model

Induction of tumor vasculature occlusion by targeting a thrombogen to newly formed blood vessels in tumor tissues represents an intriguing approach to the eradication of primary solid tumors. In the current study, we construct and express a fusion protein containing vascular endothelial growth factor (VEGF) and tissue factor (TF) to explore whether this fusion protein has the capability of inhibiting tumor growth in a colon carcinoma model. The murine cDNA of VEGF A and TF were amplified by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned into prokaryotic expression plasmid pQE30 with a linker. The expression product recombinant VEGF-TF (rVEGF-TF) was purified and proved to have comparable enzyme activity to a commercial TF and the capability of specific binding to tumor vessels. Significant decrease of tumor growth was found in the mice administered with rVEGF-TF on Day 6 after initiated rVEGF-TF treatment (P<0.05), and the tumor masses in 2 of 10 mice were almost disappeared on Day 14 after the first treatment. In addition, valid thrombogenesis and tumor necrosis were observed in the tumor tissues injected with rVEGF-TF. Our results demonstrate that occlusion of tumor vasculature with rVEGF-TF is potentially an effective approach for cancer therapy.

Evaluation of Serum and Aqueous Humor Vascular Endothelial Growth Factor in Neovascular Glaucoma

Purpose: The study aimed to evaluate and correlate between the levels of vascular endothelial growth factor (VEGF) in serum and aqueous humor in cases of neovascular glaucoma (NVG) to stand up on if it can be used as a marker for early detection of such cases. Methods: This observational case control study included 60 eyes, divided into 3 groups, group A of 30 eyes presented by cataract of different causes (not diabetic patients and no signs of NVG) as a control group and group B of 30 eyes with NVG due to different causes, group C of the same eyes in group B but after one month of treatment by intravitreal bevacizumab and laser treatment by pan retinal photocoagulation (PRP). Serum VEGF was estimated in all groups, also aqueous humor VEGF was estimated in group A and B only. In addition glycosylated hemoglobin (HbA1c) was estimated in group B;statistical analysis of the results was performed. Results: The study revealed that the commonest cause of NVG was proliferative diabetic retinopathy (PDR) in 26 cases (86.7%), HbA1c in group B revealed mean value 7.68% ± 2.75%. Serum VEFG level in the group B of cases of NVG was significantly higher than the control group A (P 0.05). Conclusions: VEGF is considered a good marker for the NVG either in serum or aqueous humor, laser treatment and the use of anti-VEGF are crucial treatment for such cases, and also glycemic control is a must for regulation of the vascular process in diabetic patients for prevention of such ocular neovascularization.

Vascular endothelial growth factor expression and significance in different grades of the Breast Imaging Reporting and Data System

Objective Mammography is the only modality proven to reduce mortality in breast cancer,and ultrasonography is a well-known adjunct to mammography screening.The Breast Imaging Reporting and Data System(BI-RADS) classification is a practical tool and is correlated with histopathology and combined use with triple assessment(examination,imaging,and biopsy) of palpable diagnostic cases.This study aimed to investigate the relationship between vascular endothelial growth factor(VEGF) expression and different grades of BI-RADS in breast cancer.Methods Ninety-six patients with breast carcinoma were evaluated using BI-RADS by ultrasonography,mammography,and a combination of both modalities.In the combined imaging assessment,BI-RADS 1-4a grade was considered when the score of ultrasonography and mammography was lower than 4a,and BI-RADS 4b-5a grade was considered when the score of ultrasonography and mammography was higher than 4a.Immunohistochemical Ultra SensitiveTM S-P method was employed to evaluate the expression of VEGF in 96 patients.Fifty patients with benign breast disease were selected as the control group.The relationship between VEGF expression and different grades of BI-RADS and that between VEGF expression and other standard prognostic parameters associated with invasive breast cancer,such as size,grade,cancer stage,and metastasis were analyzed.Results The sensitivities of ultrasonography and mammography alone was 74.0% and 84.4%,respectively;However,the sensitivity of their combination increased to 90.6%.The positive rates of VEGF in invasive breast cancer BI-RADS 4b-5(59/87,67.8%) were higher than those in BI-RADS 4a(3/9,33.3%,P<0.05) and benign breast disease tissues(BI-RADS 1-4a,11/50,22.0%)(P<0.05).There was a positive correlation between VEGF overexpression and BI-RADS 4b-5,histological grade(Ⅲ),lymph node metastasis,and distant metastasis of invasive breast cancer.VEGF expression was not related to the age and size of the tumor in each group(P>0.05).Conclusion There was a positive correlation between VEGF overexpression and BI-RADS 4b-5 grade.The overexpression of VEGF might be an important biological marker for the invasion and metastasis of breast cancer.

Vascular endothelial growth factor induced angiogenesis following focal cerebral ischemia/reperfusion injury in rabbits

BACKGROUND： Therapeutic angiogenesis has opened up new pathway for the treatment of ischemic cerebrovascular disease in recent years. The exploration of the effect of vascular endothelial growth factor （VEGF） on inducing angiogenesis following ischemia/reperfusion injury can provide better help for the long-term treatment of cerebrovascular disease in clinic. OBJECTIVE： To observe the effect of VEGF on inducing angiogenesis following focal cerebral ischemia /reperfusion injury in rabbits through the angiogenesis of microvessels reflected by the expression of the factors of vascular pseudohemophilia. DESIGN： A randomized controlled animal tria SETTNG： Department of Medical Imaging, Second Hospital of Hebei Medical University MATERIALS： Sixty-five healthy male New Zealand rabbits of clean degree, weighing （2.6±0.2） kg, aged 4.5-5 months, were used. The polyclonal antibody against vascular pseudohemophilia （Beijing Zhongshan Company）, recombinant VEGF165 （Peprotech Company, USA）, biotinylated second antibody and ABC compound （Wuhan Boster Company） were applied. METHODS： The experiments were carried out in the Laboratory of Neuromolecular Imaging and Neuropathy, Second Hospital of Hebei Medical University from May to August in 2005. （1） The rabbits were randomly divided into three groups： sham-operated group （n=15）, control group （n=25） and VEGF-treated group （n=-25）. In the control group and VEGF-treated group, models were established by middle cerebral artery occlusion （MCAO） induced focal cerebral ischemia/reperfusion. In the VEGF-treated group, VEGF165 （2.5 mg/L） was stereotactically injected into the surrounding regions of the infarcted sites immediately after the 2-hour ischemia/reperfusion; Saline of the same dosage was injected in the control group. But the rabbits in the sham-operated group were only drilled but not administrated. （2） The experimental indexes were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment respectively, 3 rabbits in the sham-operated group and 5 in the control group and VEGF-treated group were observed at each time point. The brain tissues in the surrounding regions of the infarcted sites were collected. The positive expressions of the factors of vascular pseudohemophilia in vascular endothelial cells were analyzed with immunohistochemical method. The microvessels in unit statistical field were counted with the imaging analytical software. MAIN OUTCOME MEASURES： The changes of microvascular density in the brain tissue and the positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of the infarcted sites were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment. RESULTS： All the 65 New Zealand rabbits were involved in the analysis of results without deletion. Changes of the number of microvessels at different time points in each group： There were no obvious changes at different time points in the sham-operated group. The numbers of microvessels at 7 and 14 days were obviously more in the control group than in the sham-operated group [（6.0±1.1）, （9.0±0.9） microvessels; （3.0±1.1）, （3.0±1.1） microvessels; P〈 0.05-0.01], and those at 3, 7, 14 and 28 days were obviously more in the VEGF-treated group than in the control group [（8.3±2.0）, （13.4±1.4）, （15.5±2.3）, （6.8± 1.0） microvessels; （3.4±0.6）, （6.0±1.1）, （9.0±0.9）, （3.2±0.8） microvessels; P 〈 0.01]. （2） Positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of infarcted sites： There were no obvious changes at different time points in the sham-operated group. In the control group, the changing law of the expressions was the same as that for the number of microvessels that the expression began to mildly increase at 7 days, reached the peak value at 14 days, and began to reduce at 28 days. In the VEGF-treated group, the expression was obviously increased at 3 days, also reached the peak value at 14 days, and reduced to the normal level at 70 days, but the expressions were obviously stronger than those in the control group at the same time points. CONCLUSION： Angiogenesis can be obviously induced in rabbits after the focal cerebral ischemia/reperfusion injury is treated with VEGF for 18 days.

Interleukin-1α, 6 regulate the secretion of vascular endothelial growth factor A, C in pancreatic cancer

Vascular endothelial growth factor (VEGF, namely VEGF-A) is an angiogenic polypeptide and VEGF-C is a lymphangiogenic polypeptide that has been implicated in cancer growth, invasion and metastasis. Several cytokines and growth factors play an important part in cancer progression. These cytokines and growth factors are the principal mediators of cancer cells-stromal cell interaction , which is critical for invasion of cancer cells to the surrounding tissues and metastatic dissemination to distant organs. In this study, we studied VEGF-A, C expression in cultured human pancreatic cancer cell lines and whether the presence of VEGF-A, C in the cell lines is regulated by cytokines interleukin-lct (EL-1α), and interleukin-6 (IL-6). METHODS: We used Northern blot and Western blot methods to analyze expression of the gene and protein of VEGF-A, C in all 6 tested cell lines (ASPC-1, CAPAN-1, MIA-PaCa-2, PANC-1, COLO-357 and T3M4) respectively. To analyze what is the regulator for this VEGF-A, C expression in pancreatic cancer,we used the reverse transcription -polymerase chain reaction (RT-PCR) method to analyze VEGF-A, C expression in cultured human pancreatic cancer cell lines (CAPAN-1 and COLO-357) under the stimulation with IL-1α (10μg/L) or IL-6 (100 μg/L). RESULTS:Northern blot analysis revealed the presence of the 4.1-kb VEGF-A mRNA transcript and 2.4-kb VEGF-C mRNA transcript in all 6 tested cell lines. Immunoblotting with highly specific anti-VEGF-A, anti-VEGF-C antibody revealed the presence of a molecular weight of 43-kDa VEGF-A protein and 55-kDa VEGF-C protein in all the cell lines. RT-PCR analysis revealed the levels of the VEGF-A and VEGF-C gene were 1-2 fold and a 1-fold increase in the COLO-357 cell line by stimulation with IL-la, however, no effect was found in the CAPAN-1 cell line. The levels of the VEGF-A and VEGF-C gene were 2-5 fold and a 1-fold increase in the CAPAN-1 cell line by stimulation with IL-6, but, no effect was found in the COLO-357 cell line. CONCLUSION:These findings suggested that the expression of VEGF-A, C and their regulation by IL-1α, IL-6 in pancreatic cancer contributes to the lymphatic and distant metastasis and the disease progression.

Variations in vascular endothelial growth factor levels during ovarian superovulation and reduction of ovarian hyperstimulation incidence in young women: A prospective study

Variations in Vascular Endothelial Growth Factor (VEGF) levels were prospectively evaluated in 18 young women undergoing in vitro fertilization treatments according to the “Long Protocol” and a typical pattern of VEGF levels was recorded. A significant increase in VEGF concentrations was observed only when the follicles reached a mean diameter of 15.3 mm in concurrence with mature oocyte retrieval. Since an increase in VEGF levels is related to follicular vascularity and oocyte developpment, our study supports the approach that oocyte retrieval may be performed when follicles > 15 mm in diameter appear. Anticipating egg retrieval in young patients with an optimal ovarian reserve may decrease the incidence of severe ovarian hyperstimulation, without compromising the treatment results.