Enhanced angiogenesis of porous collagen scaffolds by incorporation of TMC/DNA complexes encoding vascular endothelial growth factor

Objective To observe the expressions of microRNA-126 (miR-126) and vascular endothelial growth factor (VEGF) in gastric carcinoma tissues, and their associations with clinicopathologic features in gastric carcinoma. Methods Fifty cases of gastric carcinoma and matched tumor adjacent tissue specimens were collected from January 2012 to March 2014 in Shangqiu First People′s Hospital of He′nan Province. Expression levels of miR-126 and VEGF were examined using reverse transcriptase-polymerase chain reaction. Their correlations and the relationships between them and the clinicopathologic features in gastric carcinoma were analyzed. Results The expression levels of miR-126 in the tumor tissues and in the adjacent tissues were 0.652±0.102 and 0.379±0.069, and the expression levels of VEGF in the two kinds of tissues were 1.523±0.142 and 0.604±0.152, with statistical differences (t=8.374, P=0.001; t=29.508, P=0.000). The expressions of miR-126 and VEGF were significantly positively correlated (r=0.735, P=0.001). The expre-ssions of miR-126 and VEGF were associated with TNM stage (t=22.981, P=0.001; t=18.053, P=0.001), depth of tumor invasion (t=26.792, P=0.001; t=20.234, P=0.001) and tumor node metastasis (t=26.668, P=0.001; t=14.857, P=0.001). Conclusion miR-126 is high expressed in the gastric carcinoma tissue, which may up-regulate the expression of VEGF, and lead to the formation of new blood vessels, then to promote gastric carcinogenesis and the development process. Key words: Stomach neoplasms; MicroRNAs; Vascular endothelial growth factors

Effects of Increased Renal Tubular Vascular Endothelial Growth Factor (VEGF) on Fibrosis, Cyst Formation, and Glomerular Disease

Vascular endothelial growth factor (VEGF) is a potent mitogen and permeability factor for endothelial cells that plays a central role in angiogenesis, vascular maintenance, inflammation, and cancer. VEGF also mediates the homeostatic adaptation to hypoxic conditions by promoting an increase in vascular density to compensate for decreased oxygenation. This process is triggered by an oxygen-sensitive transcription factor, hypoxia-inducible factor-1 (HIF1alpha), which becomes active in hypoxic tissues, leading to the synthesis and secretion of VEGF. The role of HIF1alpha in other processes that involve angiogenesis such as in inflammation is less clear. Of interest, endothelial cells not only respond to but also store and secrete VEGF, which is required for the maintenance of the integrity of the vascular system. How this intracellular pool of VEGF is regulated is still not understood. Here, we found that CXCL8/IL8, a potent proangiogenic and inflammatory chemokine, up-regulates VEGF mRNA and protein levels in endothelial cells by acting on its cognate receptor, CXCR2, and that this results in the autocrine activation of VEGFR2. Surprisingly, this process does not involve HIF1alpha but instead requires the activation of the transcription factor NFkappaB. Furthermore, we identified the components of the CBM complex, Carma3, Bcl10, and Malt1, as key mediators of the CXCL8/IL8-induced NFkappaB activation and VEGF up-regulation. Together, these findings support the existence of an NFkappaB-mediated pathway by which the proinflammatory chemokine CXCL8/IL8 controls the expression of VEGF in endothelial cells, thereby promoting the activation of VEGF receptors in an autocrine fashion.

Etiology of OHSS and use of dopamine agonists

Nasal polyposis is characterized by tissue remodelling and oedematous nasal mucosa. Vascular endothelial growth factor (VEGF) plays a significant role in the regulation of remodelling in nasal polyps. TLR4 activation is associated with VEGF expression in murine macrophages and odontoblasts. This study aimed to evaluate whether lipopolysaccharide (LPS), an inducer of TLR4, stimulates VEGF expression and to determine the mechanism underlying VEGF production in nasal polyps. Nasal polyp-derived fibroblasts (NPDFs) were isolated from 10 patients with nasal polyps and exposed to LPS. LPS from Rhodobacter sphaeroides (LRS) was used to inhibit the expression levels of TLR4, MyD88 and VEGF. Messenger RNA (mRNA) expression levels of TLRs, MyD88 and VEGF were determined by gene expression microarray and semiquantitative reverse transcription-PCR. Protein expression levels of TLR4 and VEGF were analysed using western blot, immunofluorescence staining and enzyme-linked immunosorbent assay (ELISA). Activation of MAPKs (ERK, p38, and JNK) and Akt was examined using western blot analysis. The expression level of VEGF was measured by ELISA and western blot analysis in ex vivo nasal polyp organ culture. The protein expression level of VEGF was increased in nasal polyp tissues compared with inferior turbinate tissues. LRS inhibited the mRNA and protein expression of TLR4, MyD88 and VEGF in LPS-stimulated NPDFs. LPS-activated MAPKs and Akt signals, whereas MAPK inhibitors did not inhibit VEGF expression, and only Akt inhibitor blocked VEGF production. LRS reduced the production of VEGF in LPS-stimulated ex vivo organ culture. These results suggest that LPS stimulates the production of VEGF through the TLR4-Akt signalling pathway in nasal polyps. LPS may be involved in the pathogenesis of nasal polyp remodelling.

Background: Oncolytic virotherapy is a promising approach to treat cancers resistant to conventional therapies. Vaccinia virus, an enveloped DNA virus from the poxvirus family, has been used as a vector for cancer vaccines and oncolytic viral gene therapy. Wild type vaccinia virus has a natural tropism for cancer cells both in vitro and in vivo due to their increased proliferation, abnormalities in epidermal growth factor and interferon signalling, in addition to other unknown mechanisms. We have recently demonstrated that the anti-tumor efficacy of the Lister strain vaccinia virus, unlike some replicating oncolytic vectors, is not attenuated by hypoxia and even augmented 20 fold in some pancreatic ductal adenocarcinoma (PDAC) cells lines (CT Hiley et al. Gene Therapy 2009. In press). The underlying mechanism is not fully understood. Methods: Vascular Endothelial Growth Factor (VEGF) was detected by an enzyme-linked immunosorbent assay. The effect of VEGF on vaccinia virus infection in human cancer cells was investigated by manipulation of VEGF expression using siRNAs or gene over-expression. Normal human bronchial epithelial cells (NHBE) were used to assess the effect of VEGF on non-transformed cells. Results: Induction of VEGF by hypoxia was found in the supernatants of cancer cell lines showing sensitisation to vaccinia virus under hypoxia. Replication of vaccinia virus was correlated with the level of VEGF expression in a panel of human cancer cell lines. Knockdown of VEGF expression in Suit2 cells by siRNAs reduced transgene expression over 7 fold at 24 and 48 hours post infection (PI) (P<0.05 & P<0.01 respectively) and viral replication 2 fold (P<0.01) at 72 hours PI. Over-expression of VEGF in cell line MiaPaca2, which has a lower level of VEGF expression, enhanced its sensitivity to vaccinia virus. Four fold over-expression of VEGF in MiaPaca2 resulted in levels of only half that seen in Suit2 cells. However a threefold increase in viral replication was observed at 72 hours PI (P<0.01) together with significant increases in transgene expression at 24 and 48 hours and a 40% increase in cytotoxicity at 6 days PI (p<0.05) compared to control. No similar effect on vaccinia virus tropism was seen in NHBE cells at levels of VEGF expression seen in PDAC cell lines. VEGF also augmented vaccinia virus tumour targeting in vivo after systemic administration (P<0.01 at 60 hours post intravenous injection). Conclusion: This novel finding suggests that VEGF expression by cancer cells is at least partly responsible for some of the tumour tropism of wild type Lister vaccinia virus and highlights its suitability for use in poor prognosis tumour types, notably those with high levels of hypoxia and VEGF expression. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 589.

Cellular senescence prevents the proliferation of cells at risk for neoplastic transformation. Nonetheless, the senescence response is thought to be antagonistically pleiotropic and thus contribute to aging phenotypes, including, ironically, late life cancers. The cancer-promoting activity of senescent cells is likely due to secreted molecules, the identity of which remains largely unknown. Here, we have shown that senescent fibroblasts, much more than presenescent fibroblasts, stimulate tumor vascularization in mice. Weakly malignant epithelial cells co-injected with senescent fibroblasts had larger and greater numbers of blood vessels compared with controls. Accordingly, increased vascular endothelial growth factor (VEGF) expression was a frequent characteristic of senescent human and mouse fibroblasts in culture. Importantly, conditioned medium from senescent fibroblasts, more than medium from presenescent cells, stimulates cultured human umbilical vein endothelial cells to invade a basement membrane, a hallmark of angiogenesis. Increased VEGF expression was specific to the senescent phenotype and increased whether senescence was induced by replicative exhaustion, overexpression of p16(Ink4a), or overexpression of oncogenic RAS. The senescence-dependent increase in VEGF production was accompanied by very little increase in hypoxic-inducible (transcription) factor 1 alpha protein levels, and hypoxia further induced VEGF in senescent cells. This result suggests the rise in VEGF expression at senescence is not a hypoxic response. Our findings may in part explain why senescent cells stimulate tumorigenesis in vivo and support the idea that senescent cells may facilitate age-associated cancer development by secreting factors that promote malignant progression.

To investigate the clinical significance of tissue factor (TF) and vascular endothelial growth factor (VEGF) expression on peripheral blood CD14 positive monocytes in patients with diffuse large B cell lymphoma (DLBCL). The expressions of TF and VEGF on peripheral CD14+ monocytes in 41 patients with DLBCL (DLBCL group) before chemotherapy and after 4 chemotherapeutic courses, and in 20 healthy subjects (control group) were detected by flow cytometry respectively, meanwhile, the relationship of the expression of TF and VEGF with international prognostic indexes (IPI) and short-term effects were analysed. The expression levels of TF and VEGF on peripheral CD14+ monocytes in DLBCL group were significantly higher than those in control group (P<0.01), and a positive correlation was found between the two groups (r=0.755, P<0.01). The expression of TF and VEGF on CD14+ monocytes in patients with prognostic risk factors significantly increased as compared with those in patients without prognostic risk factors (P<0.05), but there were no significant differences of TF and VEGF expressions on CD14+ monocytes in DLBCL group with different sex, age, subtypes (P＞0.05). As compared with patients without prognostic risk factors, the expression levels of TF and VEGF on CD14+ monocytes of patients with prognostic risk factors significantly increased (P<0.05). The expression of TF and VEGF on CD14+ monocytes in DLBCL group showed an increasing tendency along with the increase of IPI index (P<0.01). The expression levels of TF and VEGF on CD14+ monocytes in remission group before chemotherapy were lower than those in non-remission group (P<0.01); after chemotherapy, the expression levels of TF and VEGF on CD14+ monocytes in remission group were lower than those before chemotherapy (P<0.05), while the TF and VEGF expression levels in non-remission group were no singnificauly different from TF and VEGF levels before chemtherapy (P>0.05), the survival of patients in group with low expression of TF and VEGF was superior to that in group with high expression of TF and VEGF (P<0.05). The paripheral blood CD14+ monocytes in DLBCL patients highly express the TF and VEGF, which relate with IPI, therapeutic efficacy and survival, thus the TF and VEGF expression levels are of reference significance for evaluating the therapeutic efficacy and prognosis of patients.

MIA-dependent angiogenesis and lymphangiogenesis are closely associated with progression, nodal metastasis and poor prognosis in tongue squamous cell carcinoma

Objective To observe the expression of matrix metalloproteinase(MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) in retinoblastoma (RB) and its relationship with the differentiation and optic nerve infiltration of RB. Methods Forty paraffin specimens of pathological confirmed RB were studied. They were divided into differentiated group (15 cases) and undifferentiated group (25 cases) , optic nerve infiltration group( 13 cases) and without optic nerve infiltration group(27cases). The expression of MMP-2, MMP-9 and VEGF were detected by immunohistochemistry, their relationships with the differentiation and optic nerve infiltration were also analyzed. Results The positive rate of MMP-2, MMP-9 and VEGF expression in 40 RB cases were 52.5%, 57.5% and 72.5%respectively. The expression of MMP-2, MMP-9 and VEGF in the undifferentiated group were significantly higher than those in the differentiated group (χ2= 9. 037, 9. 253, 8. 095;P＜0. 05). The expression ofMMP-2, MMP-9 and VEGF in RB with optic nerve infiltration group were significantly higher than those in RB without optic nerve infiltration group (χ2=11.045,10. 243, 8. 956;P＜0. 05). The expression of MMP-2,MMP-9 had a positive correlation with the expression of VEGF in RB (r= 0. 126, 0. 314;P ＜ 0. 05).Conclusions MMP-2, MMP-9 and VEGF expressed in RB tumor tissues. The expression of MMP-2,MMP-9 has a positive correlation with the expression of VEGF. The levels of MMP-2, MMP-9 and VEGF expression are related to optic nerve infiltration of RB cells. Key words: Retinoblastoma/pathophysiology; Matrix metalloproteinases; Vascular endothelial growth factors

Experimental abdominal aortic aneurysm growth is inhibited by blocking the JAK2/STAT3 pathway

Objective: To explore the mRNA expression of the related genes of p53, MDM2, vascular endothelial growth factor (VEGF) and hypoxia inducible transcription factors-1 a (HIF-la) in villous samples of spontaneous abortion mouse models and normal pregnancy models, and to discuss the effect of p53, MDM2 on the growth of villous trophoblast cells. Methods: The abortion-prone CBAXDBA/2 matings were established as the model of spontaneous abortion and the non-abortion-prone CBAXBALB/c matings as the model of normal pregnancy. Applied q Real-time PCR method to detect the mRNA expression levels of p53, MDM2, VEGF and HIF-la in villous samples of spontaneous abortion mouse models and normal pregnancy models. Results: The relationship of the mRNA expression level of p53, MDM2, VEGF and HIF-la in vinous samples of spontaneous abortion mouse models: in the villous samples of spontaneous abortion mouse models, the expression of p53 was positively correlated with the expression of MDM2, HIF-la (r = 0.35; r = 0.63), and the relationship was significant (P = 0.01; P < 0.001); but negatively correlated to the expression of VEGF (r = ?0.30), and the relationship was significant (P = 0.03). The expression of MDM2 was positively correlated with the expression of HIF-la (r = 0.28), and the relationship was significant (P = 0.04); and negatively correlated with the expression of VEGF (r = ?0.08), but the relationship was not significant (P = 0.57). The expression of HIF-la was negatively correlated with the expression of VEGF (r = ?0.37), and the relationship was significant (P = 0.007). The relationship of the mRNA expression level of p53,MDM2, VEGF and HIF-1 a in vinous samples of normal pregnancy models: in the vinous samples of normal pregnancy models, the expression of p53 was positively correlated with the expression of MDM2, VEGF and HIF-la (r = 0. 31; r = 0. 48; r = 0. 67), and the relationship was significant (P = 0.03; P = 0.003; P < 0.001). The expression of MDM2 was positively correlated with the expression of VEGF (r = 0. 23), but the relationship was not significant (P = 0.11); and negatively correlated with the expression of HIF-la (r = ?0.03), but the relationship was not significant (P = 0.84). The expression of HIF-la was positively correlated with the expression of VEGF (r = 0. 35), and the relationship was significant (P = 0.01). Conclusion: angiogenesis reduces in villous samples of spontaneous abortion mouse model, P53 and MDM2 involve in angiogenesis in villous samples, unlikely p53, and MDM2 have effects on normal early pregnancy villous angiogenesis and when the cell DNA damages or hypoxia exacerbates, it can induce high expression of p53, MDM2, inhibit angiogenesis in villous samples in early pregnancy. P53, MDM2 generegulate villous trophoblast cell growth by adjusting expression of HIF-1a and VEGF gene, finally influences pregnancy.

Objective To observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA (siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. Methods HaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. Results No significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group (0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α (0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. Conclusions Hypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia. Key words: Keratinocytes; Hypoxia-inducible factor, alpha subunit; RNA interference; Vascular endothelial growth factors

Vascular Endothelial Growth Factor Localization in the Adult

Objective To evaluate the expression of vascular endothelial growth factor(VEGF),basic fibroblast growth factor(bFGF)and transforming growth factor betal(TGFβ1)in non-small cell lung cancer (NSCLC)and to investigate its relationship to histopathological features.Methods The expression of VEGF,bFGF,TGFβ1 and CD34 were examined in 50 specimens of NSCLC and 18 specimens of para-cancerous tissue by using SP immunohistochemistry staining.Assessment of microvessel density(MVD)was performed.Results The positive rate of VEGF,bFGF,TGFβ1 was 68%.The expression of VEGF was related to clinical classification and lymph nodes metastasis.The difference was significant among the groups(χ2=4.535 and 4.235,P＜0.05).The expression of VEGF had no significant relationship with some pathological characteristics such as histological types and pathological grade.The positive rate of bFGF was 58%.The expression of bFGF was related to clinical classification and lymph nodes metastasis.The difference was significant among the groups (χ2=3.864 and 3.876,P＜0.05).The expression of bFGF has no significant relationship with some pathological characteristics such as histological types,pathological grade.The positive rate of TGFβ1 was 60%.The expression of TGFl3l had significant relationship with some pathological characteristics such as histological types,clinical classification and lymph nodes metastasis.The difference was significant among the groups (χ2=3.852,4.151 and 4.443,P＜0.05).The expression ofTGFβ1 was not related to pathological grade.The expression of MVD was related to clinical classification and lymph nodes metastasis.The difference was significant among the groups(t=2.312 and 2.865,P＜0.05).The expression of MVD had 110 significant relationship with some pathological characteristics such as histological types and pathological grade.The four of all factors had closed statistical significance between cancerous tissue and para.cancerous tissue.The expressions of VEGF and bFGFwere related to MVD.The difference Was significant among the groups(t=2.518 and 3.679,P＜0.05).The expression of TGFl3,had no significant relationship with MVD.Conclusion VEGF,bFGF and TGFDl have very high expressions in NSCLC.The expression levels of VEGF,bFGF and TGFβ1 are significantly related to the invasive growth and metastasis of NSCLC.VEGF and bFGF play an important role during the tumor neovascularization and tumor metastasis.TGFβ1 may be associated with the decreasing of immunoregulation function and the escape of immune survival. Key words: NSCLC; VEGF; bFGF; TGFβ1; MVD