VEGF mRNA Expression in Epithelial Ovarian Cancer: Correlation with rs699947 Gene Variant

We tested whether adenosine has differential effects on vascular endothelial growth factor (VEGF) expression under normoxic and hypoxic conditions, and whether A(1) or A(2) receptors (A(1)R; A(2)R) mediate these effects. Myocardial vascular smooth muscle cells (MVSMCs) from dog coronary artery were exposed to hypoxia (1% O(2)) or normoxia (20% O(2)) in the absence and presence of adenosine agonists or antagonists for 18 h. VEGF protein levels were measured in media with ELISA. VEGF mRNA expression was determined with Northern blot analysis. Under normoxic conditions, the adenosine A(1)R agonists, N(6)-cyclopentyladenosine and R(-)-N(6)-(2-phenylisopropyl)adenosine did not increase VEGF protein levels at A(1)R stimulatory concentrations. However, adenosine (5 microM) and the adenosine A(2)R agonist N(6)-[2-(3, 5-dimethoxyphenyl)-2-(2-methylphenyl)]ethyl adenosine (DPMA; 100 nM) increased VEGF protein levels by 51 and 132% and increased VEGF mRNA expression by 44 and 90%, respectively, in cultured MVSMCs under normoxic conditions. Hypoxia caused an approximately fourfold increase in VEGF protein and mRNA expression, which could not be augmented with exogenous adenosine, A(2)R agonist (DPMA), or A(1)R agonist [1,3-diethyl-8-phenylxanthine (DPX)]. The A(2)R antagonist 8-(3-chlorostyryl)-caffeine completely blocked adenosine-induced VEGF protein and mRNA expression and decreased baseline VEGF protein levels by up to approximately 60% under normoxic conditions but only by approximately 25% under hypoxic conditions. The A(1)R antagonist DPX had no effect. These results are consistent with the hypothesis that 1) adenosine increases VEGF protein and mRNA expression by way of A(2)R. 2) Adenosine plays a major role as an autocrine factor regulating VEGF expression during normoxic conditions but has a relatively minor role during hypoxic conditions. 3) Endogenous adenosine can account for the majority of basal VEGF secretion by MVSMCs under normoxic conditions and could therefore be a maintenance factor for the vasculature.

Objective To investigate the effects and mechanism of 17β-estradiol on the retinal neovasularization in rats with oxygen-induced retinopathy (OIR).Methods A total of 48 SD rats were randomly divided into control group A,control group B,experimental group A and experimental group B with 12 rats in each group.The rats in control group A and experimental group A received a hypodermic injection of 0.1 ml PBS,and the rats in control group B and experimental group B group received an a hypodermic injection of 0.1 ml 17β-estradiol.At postnatal day 7 (P7) and P14,the mRNA expression of vascular endothelial growth factor (VEGF) and Hypoxia-inducible factor (HIF) -1α in the retina were measured by real-time polymerase chain reaction (RT-PCR). At P14,endothelial cell nuclei breaking through the internal limiting membrane were counted after staining with hematoxylin and eosin (HE),and the protein expression of VEGF was measured after immunohistochemical staining.The changes of retinal ultrastructure were observed by transmission electron microscopy.Results At P14,the difference of the number of endothelial cell nuclei among four groups was statistically significant (F=10.7,P＜0.05).The number of endothelial cell nuclei in experimental group A was increased greater than that in control group A (P＜0.05) and experimental group B(q=5.16,P＜0.05).But there was no difference between control group A and experimental group B (q=0.25,P＞0.05).The difference of VEGF protein expression among the four groups was statistically significant (P＜0.05).Comparing experimental group A with control group A,B and experimental group B,the difference was statistically significant (P＜0.05).In experimental group A there was ganglion cell swelling,pale staining cytoplasm and mitochondria vacuolizationin,while these were normal in other three groups.At P7 and P14,the differences of VEGF and HIF-1 mRNA expression among four groups were statistically significant (F=14.7,16.1,13.4,17.5； P=0.001,0.005,0.003,0.009).At P7,the VEGF mRNA expression in control group B was more than that in control group A (q=5.22,P＜0.05).The VEGF mRNA expression in experimental group B was more than that in experimental group A (q=4.32,P＜0.05).At P14,the VEGF mRNA expression in control group B was more than that in control group A (q=3.72,P＜0.05),but there was no difference of HIF-1 mRNA expression between two groups.The VEGF and HIF-1 mRNA expression in experimental group B were both decreased more than those in experimental group A (q =5.12,4.08 ； P＜ 0.05).Conelusions 17β-estradiol has the effect of two-way regulation in VEGF mRNA,which increases VEGF expression in retina under hyperoxic conditions so as to develop the vascular system； which reduces VEGF and HIF-1αexpression so as to prevent pathologic neovascularization under hypoxic conditions. It provides some protection from the damage of retinal neovascularization. Key words: Retinal neovascularization/drug therapy; Estradiol; Vascular endothelial growth factors; Hypoxia-inducible factor 1, alpha subunit

Objective To explore the effect of vascular endothelial growth factor (VEGF) mRNA interference on glioblatoma.Methods After knockdown of VEGF mRNA expression using VEGF short hairpin RNA (shRNA),glioma U251 cell were treated with chemotherapy or radiotherapy or chemotherapy plus radiotherapy or without any treatment.The changes in cell cycle,apoptosis rate,cell colony-formation and cell morphology were observed.Results After knockdown of VEGF mRNA expression using VEGF shRNA,VEGF mRNA expression levels in glioma U251 cells were inhibited significantly (P ＜ 0.001).The flow cytometry revealed that both control cells and VEGF shRNA-transfected cells exhibited G0-G1 arrest,and reduced number of cells in the G2 and M phases.The apoptosis rate of VEGF shRNA-transfected cells was increased as compared with the control cells.It was found that various concentrations of paciltaxel reduced U251 cell viability 50％ inhibitory dose(IC50) =28.1 mg/L,and VEGF knockdown significantly sensitized U251 cells to paclitaxel (IC50 =0.02 mg/L).Groups with drug treatment alone and the radiotherapy alone showed no significant difference (P ＞ 0.05).After VEGF knockdown,colony formation in paclitaxel and radiation treated U251 cells was significantly reduced (17.57％ to 6.33％,P ＜ 0.01).Under the microscopy,it was found VEGF shRNA-transfected cells showed worse cellular morphology than non-transfected cells.Conclusion The interference of VEGF gene expression can inhibit the proliferation of U251cells,promote apoptosis of U251 cells,and increase the sensitivity of U251 cells to chemotherapy and radiotherapy.Liposomal paclitaxel can enhance drug delivery in vivo,although the VEGF gene interference and the practical application of liposomal paclitaxel remains to be tested and explored in future studies. Key words: Glioblastoma; Vascular endothelial growth factor; RNA interference ; Radiotherapy; Chemotherapy

The effect of transfection of antisense vascular endothelial growth factor (VEGF) gene on the growth of hemangioma was studied. A total of 49 cases of capillary hemangiomas of the skin were collected. Immunohistochemical method was used to detect the expression of PCNA in hemangioma tissues. According to the finding, 49 cases of hemangiomas fell into proliferating phase (27 cases) and involuting phase (22 cases) respectively. Another 5 cases of normal skin tissues adjacent to the tumor tissues served as control. Immunohistochemical staining was performed to detect the expression of VEGF in the tumor tissues and the normal tissues. The average absorbance (A) values and the average positive area rate of VEGF were measured by image analysis system (HPIAS-2000). Endothelial cells from the tumor tissues in proliferating phase were cultured. Eukaryotic expression vector was constructed by sub-cloning, and transfected into human hemangioma endothelial cells by using cation liposome as vector. The expression of VEGF mRNA and protein was detected by RT-PCR and indirect immunofluorescence assay (IFA), respectively, and the biological characteristics of the transfected endothelial cells were examined by MTT assay and flow cytometry (FCM) after transfection. Immunohistochemical results showed that the expression of VEGF in proliferating endothelial cells was remarkably higher than those in involuting endothelial cells and normal endothelial cells (P<0.01), but there was no significant difference in the expression of VEGF between involuting endothelial cells and normal ones (P>0.01). Electrophoresis and sequencing indicated that the eukaryotic expression vector containing antisense VEGF gene, i.e. pcDNA3.1-VEGF, was successfully constructed. After VEGF antisense RNA recombinant was transfected into hemangioma endothelial cells, RT-PCR revealed that the expression of VEGF mRNA in pcDNA-VEGF (V) group and blank group was obviously higher than that in pcDNA-VEGF (A) group, and that the expression of endogenous VEGF mRNA in pcDNA-VEGF (A) group was significantly inhibited. Immunohistochemical result demonstrated that, compared with blank group, there was statistically significant difference between pcDNA-VEGF (A) and pcDNA-VEGF (V) groups (P<0.01), but there was no significant difference between pcDNA-VEGF (V) group and blank group (P>0.05). The activity of endothelial cell proliferation was reduced significantly after transfection, and obvious apoptosis occurred in hemangioma endothelial cells after transfection of antisense VEGF. It was suggested that VEGF plays an important role in the pathological change of hemangiomas by promoting endothelial cell proliferation and angiogenesis. Antisense VEGF gene transfection could effectively inhibit the growth of hemanioma endothelial cells.

Aims: Serotonin 1A receptor (5-HT1A) and vascular endothelial growth factor (VEGF) are widely expressed in the neurons of the hippocampus and have significant roles in the pathophysiological processes of major depressive disorders (MDDs). The present study was designed to examine 5-HT1A and VEGF gene polymorphisms and whether the gene–gene interaction of 5-HT1A and VEGF gene variants was associated with MDD. Methods: A total of 264 MDD patients and 264 healthy controls were included in the present genetic study. The rs6295, rs1364043, and rs878567 single-nucleotide polymorphisms (SNPs) in the 5-HT1A gene and the rs699947, rs833061, and rs2010963 SNPs in the VEGF gene were selected for genotypic analyses. The generalized multifactor dimensionality reduction method was employed to assess their interactions. Results: The genotype distributions of the two genes’ respective SNPs were significantly different between patients and controls for 5-HT1A rs6295 (p = 0.041) and VEGF rs2010963 (p = 0.035); however, no significant allelic variation in 5-HT1A (rs6295, rs1364043, and rs878567) and VEGF (rs699947, rs833061, and rs2010963) was found. The interactions between 5-HT1A (rs6295, rs1364043, and rs878567) and VEGF (rs699947, rs833061, and rs2010963) had a cross-validation (CV) consistency of 10/10 and a p value of 0.0107, which was considered as the best generalized multifactor dimensionality reduction (GMDR) model. Conclusions: The interactions between 5-HT1A and VEGF gene polymorphisms may play a key role in the development of MDD in the Northern Chinese Han population.

Objective To evaluate the effect of different doses of Astragalus membranaceus on the levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in serum and lung tissues of rats with pulmonary embolism. Methods Seventy-six clean-grade healthy male Sprague-Dawley rats, aged 8-9 weeks, weighing 140-170 g, were assigned to control group (group C, n=11) and experimental group (group E, n=65) by a random number table method.The rats with pulmonary embolism in group E were further divided into 4 subgroups using a random number table method: pulmonary embolism group (group P), low-dose Astragalus membranaceus group (group H1), median-dose Astragalus membranaceus group (group H2) and high-dose Astragalus membranaceus group (group H3). The model of pulmonary embolism was established by injecting autologous blood clots into the right jugular vein.At 1 h and 1, 2, 3, 4, 5 and 6 days after successful establishment of the model, Astragalus membranaceus 20, 40 and 60 g/kg were injected intraperitoneally in H1-3 groups, respectively, while the equal volume of normal saline was given instead in group P. The chest was opened after anesthesia on day 7, and blood samples were collected from cardiac chambers for determination of concentrations of serum VEGF and bFGF by enzyme-linked immunosorbent assay.The pulmonary specimens were obtained from the upper lobe of right lungs for determination of the expression of VEGF and bFGF mRNA (using real-time polymerase chain reaction). Results Compared with group C, the concentrations of serum VEGF and bFGF were significantly increased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in the other four groups (P<0.05). Compared with group P, the serum bFGF concentration was significantly increased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in H1-3 groups (P<0.05). Compared with group H1, the serum bFGF concentration was significantly increased, the expression of VEGF mRNA and bFGF mRNA in lung tissues was up-regulated in H2 and H3 groups (P<0.05). Compared with group H2, the expression of VEGF and bFGF mRNA in lung tissues was significantly up-regulated in group H3 (P<0.05). Conclusion Astragalus membranaceus can up-regulate the expression of VEGF and bFGF in lung tissues in a dose-dependent manner, thus improving pulmonary embolism in rats. Key words: Astragalus membranaceus; Pulmonary embolism; Vascular endothelial growth factors; Fibroblast growth factor 2

Essential roles of angiotensin II in vascular endothelial growth factor expression in sleep apnea syndrome

POS-804 Donor vascular endothelial growth factor gene polymorphism association with acute allograft rejection in live related renal transplant recipient patients

ABSTRACTBackground: Preeclampsia (PE), is a pregnancy-specific complication with the placental origin which associated with altered expression of angiogenic factors. Vascular Endothelial Growth Factor (VEGF), is a growth and hypoxia-induced factor which contributes to the regulation of various processes. The present study has investigated the association of the placental VEGF −634G/C (rs2010963), −1154G/A (rs1570360), and −2549 I/D (18bpindel) polymorphisms in the promoter region with VEGF mRNA expression in PE women and control group.Methods: This case-control study was performed on the placenta of 84 PE women and 103 controls after delivery. Genotyping of the VEGF polymorphisms was done by PCR or PCR, PCR-RFLP or sequencing methods. The mRNA expression levels were measured by Quantitative Real-Time PCR.Results: The relative mRNA expression of VEGF gene was significantly higher in PE women compared to controls. The relative mRNA expression of VEGF gene was significantly higher in women with −634CC genotype compared to CG + GG genotypes in PE women and total studied women but not in control women. However, there was no association between the placental VEGF −1154G/A and −2549 I/D polymorphisms and VEGF mRNA expression neither in PE nor in control groups.Conclusion: The current study found higher mRNA expression of placental VEGF gene in PE women. The mRNA expression of the placental VEGF gene has been up-regulated in the placenta of women with −634CC genotype. No association was found between the placental VEGF −1154G/A and −2549 I/D polymorphisms and VEGF mRNA expression.

VEGF, FGF2, TGFB1 and TGFBR1 mRNA expression levels correlate with the malignant transformation of the uterine cervix

We have quantified the expression of all 4 isoforms of vascular endothelial growth factor (VEGF) mRNA in non-small-cell lung cancer (NSCLC) using a new kinetic quantitative PCR method, real-time quantitative (RTQ) RT-PCR, and investigated the association between VEGF expression at the mRNA and protein levels and the clinicopathologic variables, tumor angiogenesis, patient survival and timing of relapse. Surgical tumor specimens from 72 NCSLC patients (37 squamous-cell carcinomas, 35 adenocarcinomas) were examined. Twenty-eight patients had stage I, 10 stage II and 34 stage IIIA or IIIB disease. Total VEGF mRNA (all 4 isoforms) was quantified by RTQ RT-PCR, while VEGF protein expression and microvessel number in tumors were assessed immunohistochemically. VEGF mRNA was detected in all 72 tumor samples at significantly higher levels than in adjacent normal tissue. Tumoral VEGF mRNA levels correlated strongly with the VEGF protein staining score and microvessel count. Adenocarcinomas showed significantly higher VEGF mRNA expression and a higher protein staining score than squamous-cell carcinomas. High tumoral VEGF mRNA expression was associated with advanced (IIIA or IIIB) tumor stage, lymph node metastasis, high tumoral microvessel counts, short patient survival (<24 months) and early relapse (<12 months), while a high VEGF protein staining score was associated with high tumoral microvessel counts, short patient survival and early relapse. Patients with high tumoral levels of both VEGF mRNA and protein had significantly shorter survival and earlier relapse. In multivariate analysis, the VEGF protein staining score and nodal status were the most important independent predictors of survival and recurrence. We conclude that RTQ RT-PCR is a sensitive method for detecting and quantifying VEGF mRNA expression in NSCLC and that the expression levels of total VEGF mRNA and protein in NSCLC are strongly associated with histologic type, tumor angiogenesis, survival and timing of relapse. High VEGF expression in adenocarcinomas may contribute to their greater metastatic potential.

Cyclopentenone-prostaglandin derivatives, including the peroxisome-proliferator activated receptor gamma (PPARgamma) ligand 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2), inhibit tumor cell growth in vitro and in vivo. As 15d-PGJ2 was found to stimulate the expression of vascular endothelial growth factor (VEGF) in endothelial cells, we investigated whether 15d-PGJ2 induces this angiogenic factor in the human androgen-independent PC 3 prostate and the 5637 urinary bladder carcinoma cell line. In PC 3 cells, 15d-PGJ2 caused a dose-dependent increase in VEGF mRNA expression, as determined by RT-PCR. Stimulation started after 6 h, and after 72 h, VEGF mRNA expression reached a maximum of 3.3+/-0.3 U, 4.4+/-0.3 U and 6.1+/-0.1 U with 1, 5 and 10 microM 15d-PGJ2, respectively. Between 12-72 h, VEGF protein production was stimulated by up to 2-fold with 5 and 10 microM 15d-PGJ2 as assessed by ELISA in PC 3 cell-conditioned medium. In 5637 cells, 15d-PGJ2 did not alter VEGF mRNA expression for up to 72 h. Thereafter, VEGF mRNA expression was transiently increased from 2.3+/-0.8 U in control cells to 4.6+/-0.5 U in 1 microM and 5.9+/-0.6 U in 5 microM 15d-PGJ2-treated cells. VEGF protein production was only moderately stimulated (1.7-fold). 10 microM 15d-PGJ2 had no effect on VEGF mRNA expression in 5637 cells, but effectively reduced viability in both cell lines. 15d-PGJ2 also increased PPARgamma mRNA expression in both cell lines. While in PC 3 cells, stimulation of PPARgamma mRNA expression occurred after 72 h, in 5637 cells, a transient stimulation took place after 6 h (4-fold). We demonstrated that 15d-PGJ2 induces VEGF in PC 3 and 5637 cancer cells. This might be important if PG-analogues are considered as antitumor agents.

Neoangiogenesis is essential for successful wound repair. Platelets are among the earliest cells recruited to a site of skeletal injury and are thought to provide numerous factors critical to successful repair. The release of platelet-derived growth factor (PDGF) after skeletal injury increases osteoblast proliferation, chemotaxis, and collagen synthesis; however, its angiogenic effect on osteoblast biology remains unknown. The purpose of this study was to investigate the effect of recombinant human (rh)PDGF-BB on the synthesis of vascular endothelial growth factor (VEGF) by primary neonatal rat calvarial osteoblasts. Furthermore, the authors investigated whether PDGF works in concert with hypoxia, another component of the fracture microenvironment, to additively or synergistically induce VEGF production. Osteoblast cultures were stimulated with varying concentrations of rhPDGF-BB (1, 10, 50, and 100 ng/ml) in normoxic and hypoxic (<1% oxygen) conditions for 0, 3, 6, 12, and 24 hours, and VEGF gene expression was analyzed by Northern blot analysis. To determine whether rhPDGF-BB-induced VEGF messenger RNA (mRNA) expression was transcriptionally mediated or required de novo protein synthesis, transcription, and translation, studies were performed using actinomycin D and cycloheximide, respectively. Treatment with 50 ng/ml rhPDGF-BB resulted in a 2.4-fold increase in VEGF mRNA expression after 3 hours. Interestingly, rhPDGF-BB and hypoxia seemed to have an additive effect, resulting in a 3.7-fold increase in VEGF mRNA expression after 6 hours in primary neonatal rat calvarial osteoblasts. Furthermore, by using actinomycin D and cycloheximide, the authors demonstrated that the rhPDGF-BB-induced VEGF mRNA expression was transcriptionally mediate and not dependent on de novo protein synthesis. These data demonstrate that rhPDGF-BB transcriptionally increases osteoblasts VEGF mRNA expression in vitro. Furthermore, the semiquantitative results suggest that rhPDGF-BB and hypoxia act additively to increase VEGF mRNA expression. It is postulated that similar mechanisms may occur in vivo, at a site of skeletal injury, to induce neoangiogenesis and promote fracture repair.

To observe the effect of Biantie (bian stone plaste) pretreatment on serum level of prolyl hydroxylase domain 2 (PHD2) and hypoxia-inducible factor-1α (HIF-1α) in rats with acute hypobaric hypoxia induced-brain injury, and to explore the possible mechanism of Biantie on preventing brain injury at high altitude. Forty-five male SD rats were randomly divided into a blank group, a model group, a Biantie group, a medication group and a Biantie+inhibitor group, 9 rats in each group. The rats in the Biantie group the and the Biantie+inhibitor group were pretreated with Biantie at "Taiyuan" (LU 9), "Neiguan" (PC 6) and "Renying" (ST 9), 2 h each time, once a day; the rats in the medication group were treated with intragastric administration of rhodiola capsule solution (280 mg/kg) for 14 d; the rats in the Biantie+inhibitor group were intraperitoneally injected with the PHD inhibitor dimethyloxalyl glycine (DMOG) at a dose of 40 mg/kg 24 h before the establishment of the model. After the intervention, except for the blank group, the rats in the remaining 4 groups were placed in the oxygen chamber to simulate a high-altitude environment to establish the acute hypobaric hypoxia brain injury model. The arterial blood-gas analysis indexes [blood oxygen saturation (SaO2), lactic acid (Lac), blood sodium (Na+), blood potassium (K+)] and brain water content were detected in each group; the histomorphology of cerebral cortex was observed by HE staining; the serum levels of PHD2 and HIF-1α as well as vascular endothelial growth factor (VEGF) were detected by ELISA; the VEGF protein expression in brain tissue was detected by Western blot; the VEGF mRNA expression in brain tissue was detected by real-time fluorescent quantitative PCR. Compared with the blank group, the levels of SaO2 and Na+ in the model group were decreased (P<0.05), while the levels of Lac and K+ as well as the water content of brain tissue were increased (P<0.05). Compared with the model group, the level of SaO2 in the Biantie group and the medication group was increased (P<0.05), while the levels of Lac, K+ and the water content of brain tissue were decreased (P<0.05); the level of Na+ in the Biantie group was increased (P<0.05). Compared with the Biantie group, the level of SaO2 in the Biantie+inhibitor group was decreased (P<0.05), and the level of Lac and the water content of brain tissue were increased (P<0.05). In the model group, the cortical tissue cells were loose and disordered, the cortical blood vessels were dilated, and the cells were obviously swollen; the anoxic injury in the Biantie group and the medication group was lighter, and the anoxic injury in the Biantie+inhibitor group was more obvious than that in the Biantie group. Compared with the blank group, the serum PHD2 content in the model group was decreased and the HIF-1α content was increased (P<0.05), and the content of VEGF in serum and VEGF protein and mRNA expressions in brain were increased (P<0.05). Compared with the model group, the content of PHD2 in serum in the Biantie group and the medication group was increased (P<0.05), and the level of HIF-1α was decreased (P<0.05), and the content of VEGF in serum as well as VEGF protein and mRNA expressions in brain were decreased (P<0.05). Compared with the Biantie group, the serum PHD2 content in the Biantie+inhibitor group was decreased and HIF-1α level were increased (P<0.05), and the content of VEGF in serum as well as VEGF mRNA expression in brain were increased (P<0.05). Biantie at "Taiyuan" (LU 9), "Neiguan" (PC 6) and "Renying" (ST 9) could regulate serum PHD2/HIF-1α to down-regulate VEGF expression, reduce brain edema and enhance anti-hypoxia ability, so as to achieve the purpose of preventing brain injury at high altitude.

Vascular endothelial growth factor (VEGF) is a potent and specific mitogen for vascular endothelial cells and promotes neovascularization in vivo. To determine whether interleukin-1 beta (IL-1 beta), which is present in atherosclerotic lesions, induces VEGF gene expression in vascular smooth muscle cells, we performed RNA blot analysis on rat aortic smooth muscle cells (RASMC) with a rat VEGF cDNA probe. IL-1 beta increased VEGF mRNA levels in RASMC in a time- and dose-dependent manner. As little as 0.1 ng/ml IL-1 beta increased VEGF mRNA levels by 2-fold and 10 ng/ml IL-1 beta increased VEGF mRNA by 4-fold. We also measured the half-life of VEGF mRNA and performed nuclear run-on experiments before and after addition of IL-1 beta to see if IL-1 beta increased VEGF mRNA levels by stabilizing the mRNA or by increasing its rate of transcription. The normal, 2-h half-life of VEGF mRNA in RASMC was lengthened to 3.2 h (60%) by IL-1 beta, and IL-1 beta increased the rate of VEGF gene transcription by 2.1-fold. In immunoblot experiments with an antibody specific for VEGF, we found that IL-1 beta increased VEGF protein levels in RASMC by 3.3-fold. Together these data indicate that IL-1 beta induces VEGF gene expression in smooth muscle cells. This IL-1 beta-induced expression of VEGF may accelerate the progression of atherosclerotic lesions by promoting the development of new blood vessels.