Endothelial Growth Factor Gene Expression Research Articles

Expression of Vascular Endothelial Growth Factor and Anti-Proliferative Activity of Flaxseed Oil Alone and In Combination with Mefenamic Acid in Cell Lines

The flaxseed oil contains alpha-linolenic acid, lignans and flavonoids. It possesses anti-inflammatory, antioxidant, and tumor-suppressive effects. The purpose of this research was to explore the anti-proliferative effect of flaxseed oil alone and flaxseed oil in combination with mefenamic acid on human umbilical vein endothelial cells and Kaposi sarcoma cells and to identify the vascular endothelial growth factor gene expression. The anti-proliferative effects of flaxseed oil alone and in combination with mefenamic acid were investigated in vitro by the 3-(4, 5-Dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The effects of flaxseed oil alone and its combination with mefenamic acid were tested on the expression of vascular endothelial growth factor genes using real-time polymerase chain reaction. The results showed that the IC50 of flaxseed oil for human umbilical vein endothelial cells was 29.72 µg/ml and for Kaposi sarcoma cells was 16.48 µg/ml; moreover, the IC50 for flaxseed oil in combination with mefenamic acid for human umbilical vein endothelial cells was 252 µg/ml and for Kaposi sarcoma cells was 147.6 µg/ml. These outcomes revealed that flaxseed oil alone and its combination with mefenamic acid possess anti-angiogenic activity in a concentration-dependent manner which supported by significant decline in vascular endothelial growth factor expression.

The Supression of Migration and Metastasis via Inhibition of Vascular Endothelial Growth Factor in Pancreatic Adenocarcinoma Cells Applied Danusertib.

Pancreatic ductal adenocarcinoma is an extremely deadly type of cancer with a high metastatic potential. Genetic factors in cellular events play an important role in the emergence of this situation. One of these factors is Aurora kinase family members, which play a role in migration, invasion, and cell cycle. In this study, the expression of vascular endothelial growth factor gene, which plays a role in migration, metastasis, and angiogenesis, on cystic fibrosis human pancreatic ductal adenocarcinoma 1 cells of danusertib, a pan-Aurora kinase inhibitor, was examined. The half maximal inhibitory concentration (IC50) value (400 nM) of danusertib in cystic fibrosis human pancreatic ductal adenocarcinoma 1 cells was determined by the wound-healing test depending on the dose and time and migration with CIM-Plate 16 in the xCELLingence system. In addition, the effect of danusertib on migration was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) method and vascular endothelial growth factor gene expression. When the dose- and time-dependent danusertib-applied cystic fibrosis human pancreatic ductal adenocarcinoma 1 cells were compared with the control group, it was observed that the wound formed did not close. In the xCELLigence system CIM-Plate 16 migration analysis, it was observed that migration was inhibited in the group administered danusertib in parallel with the wound dehiscence experiment. The gene expressions of vascular endothelial growth factor decreased 0.5-fold at the 24th hour and 0.3-fold at the 48th hour in the Danusertib-administered groups. Danusertib, a pan-Aurora kinase inhibitor, is predicted to be used as a potential agent in pancreatic cancers due to its antitumor and anti-metastatic effect.

Nanodiamonds inhibit the proliferation and migration of endothelial cells in a tumor/endothelial cells co-culture microfluidic system

The effect of nanoparticles on co-cultured tumor and endothelial cells has not been consistently elucidated. To address this question, a microfluidic chip for the co-culture of tumor and endothelial cells is fabricated herein. Human umbilical vein endothelial cells (HUVECs) and human cervical cancer cells (Hela) are respectively cultured in the upper and lower chambers of the chip, and the culture medium with carboxylated nanodiamonds (NDs) is applied to HUVECs by perfusion. The results show that, compared to the individually cultured HUVECs, the HUVECs co-cultured with Hela exhibit cancerous characteristics, with increased leakiness, migration, and proliferation. It is also found that, while the presence of NDs can lead to increased endothelial permeability, the migration and proliferation of the co-cultured HUVECs are reduced, and the co-cultured HUVECs are hindered, to some extent, from becoming cancerous. The results of gene analysis also prove that NDs can inhibit the proliferation of HUVECs by significantly down-regulating the vascular endothelial growth factor (VEGF) gene expression. The mechanism of suppressing tumor growth by inhibiting tumor vascular proliferation was also demonstrated in-vivo using a tumor-bearing mouse model.

Analysis of ANRIL Isoforms and Key Genes in Patients with Severe Coronary Artery Disease.

ANRIL (Antisense Noncoding RNA in the INK4 Locus), also named CDKN2B-AS1, is a long non-coding RNA with outstanding functions that regulates genes involved in atherosclerosis development. ANRIL genotypes and the expression of linear and circular isoforms have been associated with coronary artery disease (CAD). The CDKN2A and the CDKN2B genes at the CDKN2A/B locus encode the Cyclin-Dependent Kinase inhibitor protein (CDKI) p16INK4a and the p53 regulatory protein p14ARF, which are involved in cell cycle regulation, aging, senescence, and apoptosis. Abnormal ANRIL expression regulates vascular endothelial growth factor (VEGF) gene expression, and upregulated Vascular Endothelial Growth Factor (VEGF) promotes angiogenesis by activating the NF-κB signaling pathway. Here, we explored associations between determinations of the linear, circular, and linear-to-circular ANRIL gene expression ratio, CDKN2A, VEGF and its receptor kinase insert domain-containing receptor (KDR) and cardiovascular risk factors and all-cause mortality in high-risk coronary patients before they undergo coronary artery bypass grafting surgery (CABG). We found that the expression of ANRIL isoforms may help in the prediction of CAD outcomes. Linear isoforms were correlated with a worse cardiovascular risk profile while the expression of circular isoforms of ANRIL correlated with a decrease in oxidative stress. However, the determination of the linear versus circular ratio of ANRIL did not report additional information to that determined by the evaluation of individual isoforms. Although the expressions of the VEFG and KDR genes correlated with a decrease in oxidative stress, in binary logistic regression analysis it was observed that only the expression of linear isoforms of ANRIL and VEGF significantly contributed to the prediction of the number of surgical revascularizations.

Evaluation of Vascular Endothelial Growth Factor Gene Expression in Recellularized Liver Tissue by Mouse Embryo Fibroblast.

The aim of the present study was to evaluate alterations in the vegf gene expression as an angiogenic factor in mouse embryo fibroblasts seeded on the decellularized liver fragments. Liver tissue samples (n = 10) collected from adult male mice were randomly divided into decellularized and native control groups. Tissues were decellularized by treating with 1% Triton X-100 and 0.1% SDS for 24 hours and assessed by H&E staining and SEM. Then DNA content analysis and toxicity tests were performed. By centrifugation, DiI-labeled mouse embryo fibroblasts were seeded on each scaffold and cultured for one week. The recellularized scaffolds were studied by H&E staining, SEM, and LSCM. After RNA extraction and cDNA synthesis, the expression of the vegf gene in these samples was investigated using real-time RT-PCR. Our observations showed that the decellularized tissues had morphology and porous structure similar to the control group, and their DNA content significantly reduced (p < 0.05) and reached to 4.12% of the control group. The MTT test indicated no significant cellular toxicity for the decellularized scaffolds. Light microscopy, SEM, and LSCM observations confirmed the attachment and penetration of embryonic fibroblast cells on the surface and into different depths of the scaffolds. There was no statistically significant difference in terms of vegf gene expression in the cultured cells in the presence and absence of a scaffold. The reconstructed scaffold had no effect on vegf gene expression. Decellularized mouse liver tissue recellularized by embryonic fibroblasts could have an application in regenerative medicine.

Cooling of male rat skeletal muscle during endurance‐like contraction attenuates contraction‐induced PGC‐1α mRNA expression

AbstractThis study aimed to determine effects of cooling on contraction‐induced peroxisome proliferator‐activated receptor γ coactivator‐1α (PGC‐1α) and vascular endothelial growth factor (VEGF) gene expression, phosphorylations of its related protein kinases, and metabolic responses. Male rats were separated into two groups; room temperature (RT) or ice‐treated (COLD) on the right tibialis anterior (TA). The TA was contracted isometrically using nerve electrical stimulation (1‐s stimulation × 30 contractions, with 1‐s intervals, for 10 sets with 1‐min intervals). The TA was treated before the contraction and during 1‐min intervals with an ice pack for the COLD group and a water pack at RT for the RT group. The muscle temperature of the COLD group decreased to 19.42 ± 0.44°C (p &lt; 0.0001, −36.4%) compared with the RT group after the experimental protocol. An increase in mRNA expression level of PGC‐1α, not VEGF, after muscle contractions was significantly lower in the COLD group than in the RT group (p &lt; 0.0001, −63.0%). An increase in phosphorylated AMP‐activated kinase (AMPK) (p = 0.0037, −28.8%) and a decrease in glycogen concentration (p = 0.0231, +106.3%) after muscle contraction were also significantly inhibited by cooling. Collectively, muscle cooling attenuated the post‐contraction increases in PGC‐1α mRNA expression coinciding with decreases in AMPK phosphorylation and glycogen degradation.

Ameliorative effect of hesperidin against high dose sildenafil-induced liver and testicular oxidative stress and altered gene expression in male rats

BackgroundThe clinical use of sildenafil citrate (Viagra), a drug used to treat erectile dysfunction, is limited because of its many side effects on tissues. In this context, we aimed to investigate the protective effects of hesperidin, a citrus flavonoid, on hepatic and testicular damage induced by a high dose of sildenafil citrate in male rats. Rats were randomly divided into four groups. The first group was used as the control group. The second group was orally administered sildenafil citrate at a high dose of 75 mg/kg thrice a week. In the third group, hesperidin was administered orally at a dose of 50 mg/kg/day. The fourth group was administered 75 mg/kg sildenafil citrate three times a week with 50 mg/kg hesperidin daily. The experiment lasted for 28 days.ResultsIn the sildenafil-treated groups, blood indices were altered, liver function tests were deranged, and serum testosterone levels were reduced. In the liver and testicular tissue, sildenafil citrate treatment resulted in significant reductions in catalase and total antioxidant capacity; as well as increased malondialdehyde, reactive oxygen species, and nitrous oxide levels. In addition, sildenafil citrate treatment caused abnormal histopathological patterns in both the liver and the testes. Liver vascular endothelial growth factor and testicular steroidogenic acute regulatory protein gene expression were upregulated.ConclusionsHesperidin attenuated the harmful effects of intensive sildenafil citrate treatment on liver and testicular functions, alleviated oxidative stress and normalized blood indices. Therefore, hesperidin could be protective against sildenafil citrate-induced oxidative damage that may develop over the long term.

Are VEGF and SCUBE1 gene expressions increased in diabetic retinopathy?

Abstract Objectives In the current study, we synergistically evaluated vascular endothelial growth factor (VEGF) gene expression levels and signal peptide-CUB-EGF domain-containing protein 1 (SCUBE1) gene expression levels in diabetic patients without retinopathy, non-proliferative diabetic retinopathy (NPDR), and proliferative diabetic retinopathy (PDR). Methods 94 blood samples from 26 healthy controls, 29 non-DR, 22 NPDR, and 17 PDR patients were collected in sterile EDTA tubes. Total RNA was obtained from these samples without waiting and then converted to cDNA. The expression levels of the VEGF and SCUBE1 genes were determined by quantitative real-time polymerase chain reaction (qPCR). Results SCUBE1 gene expression levels were 2.15 (p=0.015), 1.75 (p=0.799), 2.37 (p=0.037) times higher, and VEGF gene expression levels were 1.71 (p=0.023), 1.75 (p=0.012), 1.85 (p=0.031) times higher in the non-DR, NPDR, and PDR groups compared to the control group, respectively. VEGF gene expression levels were significantly higher in participants with HbA1c levels ≥5.7% compared to those with &lt;5.7. SCUBE1 and VEGF gene expression levels were significantly higher in participants with fasting plasma glucose (FPG) levels ≥126 mg/dL than those with &lt;126 mg/dL. Conclusions As a result, SCUBE1 gene expression levels are higher than VEGF gene expression levels, especially in the PDR group. Therefore, SCUBE1 may contribute to the pathology of DR just like VEGF by generating angiogenesis. However, we believe there is a need for experimental animal model studies with DR examining SCUBE1 gene expression levels in tissue samples.

Antihelminthic and antiangiogenic effects of zinc oxide nanoparticles on intestinal and muscular phases of trichinellosis.

Trichinellosis is a worldwide zoonotic disease affecting a wide range of mammals, including humans. It has intestinal and muscular phases. The current work was done to experimentally evaluate the efficacy of zinc oxide nanoparticles (ZnO NPs) and their combination with albendazole on intestinal and muscular stages of Trichinella spiralis (T. spiralis) infection. We had five main groups of mice: Group 1, non-infected control; Group 2, infected control; Group 3, infected and treated with albendazole; Group 4, infected and treated with ZnO NPs; and Group 5, infected and treated with albendazole and ZnO NPs. Each group was divided into two subgroups (A for the intestinal phase and B for the muscular phase). Drug effects were evaluated by parasitological, histopathological, and biochemical studies, including oxidant/antioxidant analysis and vascular endothelial growth factor (VEGF) gene expression in muscle tissue by quantitative real-time PCR. ZnO NPs resulted in a significant reduction of both intestinal and muscular phases of T. spiralis. Their combination with albendazole resulted in the complete eradication of adult worms and the maximum reduction of larval deposition in muscle tissue. Additionally, the treatment showed improvement in T. spiralis-induced pathological changes and oxidative stress status. Moreover, a significant decrease in VEGF gene expression was detected in the treated groups when compared with the infected control. In conclusion, ZnO NPs presented an antihelminthic effect against both adult and larval stages of T. spiralis. In addition, it enhanced antioxidant status and suppressed angiogenesis in muscle.

MCP-1 facilitates VEGF production by removing miR-374b-5p blocking of VEGF mRNA translation

Monocyte chemotactic protein-1 (MCP-1) is known to be able to facilitate vascular endothelial growth factor (VEGF) gene expression, hence promoting vascular hyperpermeability and neovascularization. We show here that a microRNA molecule, miR-374b-5p can target the 3′-untranslated region of the VEGF mRNA, thus preventing VEGF production. Additionally, MCP-1 promotes the acetylation of transcription factor stat3 at Lys685, which facilitates the formation of an ac-stat3-DNA methyltransferase-histone methyltransferase complex (ac-stat3/DNMT1/EZH2) that binds to the promoter of the miR-374b-5p gene. This results in diminished miR-374b-5p expression and enhanced VEGF production. Moreover, treatment of appropriate animal models either with a miR-374b-5p mimicry or with inhibitors of either stat3 acetylation, DNMT1, or EZH2, leads to marked inhibition of MCP-1-promoted neovascularization and tumor growth. These findings indicate that MCP-1 facilitated inhibition of miR-374b-5p gene expression leads to the removal of a block of VEGF mRNA translation by miR-374b-5p. This mechanism could be of importance in the modulation of inflammatory conditions.

Angiogenic T Cells: Potential Biomarkers for the Early Diagnosis of Interstitial Lung Disease in Autoimmune Diseases?

(1) Background: We explored, for the first time, the contribution of angiogenic T cells (TAng) in interstitial lung disease associated to autoimmune disease (AD-ILD+) as potential biomarkers of the disease, evaluating their role in the underlying vasculopathy and lung fibrosis. Additionally, the relationship of TAng with clinical manifestations and cellular and molecular endothelial dysfunction-related biomarkers was assessed. (2) Methods: We included 57 AD-ILD+ patients (21 with rheumatoid arthritis (RA)-ILD+, 21 with systemic sclerosis (SSc)-ILD+ and 15 with other AD-ILD+) and three comparative groups: 45 AD-ILD− patients (25 RA-ILD− and 20 SSc-ILD−); 21 idiopathic pulmonary fibrosis (IPF) patients; 21 healthy controls (HC). TAng were considered as CD3+CD184+CD31+ by flow cytometry. (3) Results: A similar TAng frequency was found between AD-ILD+ and IPF, being in both cases lower than that observed in AD-ILD− and HC. A lower TAng frequency was associated with negative Scl-70 status and lower FEV1/FVC ratio in SSc-ILD+, as well as with men in RA-ILD+ and non-specific interstitial pneumonia radiological pattern in other AD-ILD+. No relationship between TAng and endothelial progenitor cells, endothelial cells and vascular endothelial growth factor gene expression and protein levels was disclosed. (4) Conclusions: Our findings suggest TAng as potential biomarkers for the early diagnosis of ILD in AD.

Thymosin‐β4, and Human Vitronectin peptides Grafted to Collagen Tune Adhesion or VEGF Gene Expression in Human Cell Lines**

AbstractIn order to improve collagen bioactivity for regenerative medicine approaches, thymosin‐β4 (Tβ4P) and Human Vitronectin (HVP) derived peptides are grafted to collagen by thiol‐ene Michael addition. Tβ4P and HVP are known to exert a pro‐angiogenic and a pro‐adhesive activity respectively and HVP is involved in osteogenesis promotion. The ability of these peptides to increase collagen cell adhesion and angiogenesis properties is assessed on human cell lines. In particular, HVP‐grafted collagen increased human osteoblast adhesion and cell proliferation: after 24 h, both adhesion and proliferation roughly showed a 4‐fold increase, if compared to pristine collagen. Tβ4P‐grafted collagen promotes Vascular Endothelial Growth Factor (VEGF) gene expression in human vascular cell lines by more than 7 times. These results suggest that HVP‐grafted collagen may be an interesting biomaterial for bone tissue regeneration, while Tβ4P‐grafted collagen is useful for angiogenesis promotion.

The role of melatonin in angio-miR-associated inhibition of tumorigenesis and invasion in human glioblastoma tumour spheroids

Micro-RNA (miRNA)-based regulation of hypoxia, angiogenesis and tumour growth provides promising targets for effective therapy in malignant glioblastoma multiforme (GBM). Accumulating evidence suggests a potential role of melatonin in miRNA expression in cancer cells. Despite these findings, the melatonin–miRNA interaction in GBM and the effect of this interaction on GBM tumour development and invasion are not clearly understood. The aim of the present study was to evaluate the effects of melatonin on human GBM tumour spheroid tumorigenesis and invasion in vitro, and to analyse the interaction between 36 angio-miRNAs and the HIF1/VEGF/MMP9 axis, which is known to be associated with the antitumour effect of melatonin. We found that melatonin is able to selectively induce cell death in single-layer U87-MG cells (a GBM cell line) in a dose- and time-dependent manner, as characterized by MTT assay. The use of tumour spheroids and a Matrigel invasion assay revealed that melatonin impairs tumorigenesis, and it significantly reduced both the tumour spheroid area and invasion rate, especially at the 0.5 mM and 1 mM concentrations. This inhibition was accompanied by strong reductions in hypoxia-inducible factor 1-α (HIF1-α) and vascular endothelial growth factor (VEGF) gene expression and protein levels in GBM tumour spheroids. In addition, melatonin significantly reduced the relative gene expression and protein levels of matrix metalloproteinase-9 (MMP-9). This study revealed that six differentially expressed angio-miRs (miR-15b, miR-18a-5p, miR-23a-3p, miR-92a-3p, miR-130a-5p, miR-200b-3p) may play important roles in GBM tumorigenesis and invasion, and all respond to melatonin therapy. Our results suggest that melatonin inhibits tumorigenesis and invasion of human GBM tumour spheroids, possibly by suppressing HIF1-α/VEGF/MMP9 signalling via regulation of angio-miRNAs.

Effects of silencing epididymal vascular endothelial growth factor (VEGF) expression on hyaluronidase (HYD) activity in arsenic poisoning rats through downregulating VEGF receptor 2 (VEGFR2)

ABSTRACT RNA interference (RNAi) was used to investigate the role of epididymal vascular endothelial growth factor (VEGF) gene expression on sperm hyaluronidase (HYD) in a rat model of arsenic poisoning and to identify a new gene therapy target for male infertility caused by arsenic poisoning. The Rat model of chronic arsenic poisoning was established. And we found that positive expression of VEGF and VEGF receptor 2 (VEGFR2) was observed by Immunohistochemical staining in the epididymal tissues of arsenic-exposed rats. Subsequently, VEGF-shRNA-1, VEGF-shRNA-2 and VEGF shRNA-3 expression vectors containing epididymal VEGF-shRNA lentivirus were constructed and injected into the bilateral epididymis of each group of rats (Control group, NC-shRNA negative infection group, VEGF-shRNA-1 group, VEGF-shRNA-2 group, VEGF-shRNA-3 group) (n = 10 per group). Compared with the negative infection group and the normal control group, the expression of VEGF and VEGFR2 mRNA and protein levels were significantly decreased following epididymal infection. In addition, the HYD activity was all significantly lower than that in the normal control group and the negative infection group. Taken together, epididymal VEGF gene silencing may inhibit the activity of sperm HYD through downregulating VEGFR2.

Investigation of drugs affecting hypertension in bevacizumab-treated patients and examination of the impact on the therapeutic effect.

BackgroundIn patients treated with bevacizumab, hypertension may be a biomarker of therapeutic efficacy. However, it is not clear whether drugs that control blood pressure influence bevacizumab's efficacy. In this study, we investigated drugs that may affect hypertension in bevacizumab‐treated patients and examined the impact on the therapeutic effect.Patients and methodsWe analyzed 3,724,555 reports from the third quarter of 2010 to the second quarter of 2015. All data were obtained from the Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) analysis. In this retrospective cohort study, we investigated a total of 58 patients diagnosed with colorectal cancer and treated for the first time with bevacizumab containing XELOX or mFOLFOX6 at The University of Tokushima Hospital between January 2010 and December 2015. The effect of the treatment was evaluated according to Response Evaluation Criteria in Solid Tumors version 1.0. Thereafter, the effect was confirmed using Gene Expression Omnibus (GEO) and cultured cells.ResultsThere are few reports in FAERS of hypertension in patients treated with omeprazole on bevacizumab. Based on the chart review, patients who used proton pump inhibitors (PPI) had a lower response to treatment than those who did not (response rate: 25% vs 50%). Furthermore, experiments on GEO and cell lines suggested that induction of vascular endothelial growth factor (VEGF) gene expression by PPIs is the cause of the reduced therapeutic effect.ConclusionPPIs prevent hypertension in bevacizumab‐treated patients but may reduce bevacizumab's anti‐tumoral effects by inducing VEGF expression.

The Effect of Aerobic Training on Tumor Necrosis Factor alpha, Hypoxia-Inducible Factor-1 alpha &amp; Vascular Endothelial Growth Factor Gene Expression in Cardiac Tissue of Diabetic Rats

Objective: The goal of this research was to determine the influence of 4 weeks aerobic training on gene expression of tumor necrosis factor alpha (TNF-α), hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in the cardiac tissue of diabetic rats.&#x0D; Materials and Methods: In an experimental study, 30 male wistar rats were partitioned into three groups (n=10), diabetic aerobic training, diabetic control and healthy control (n=10). Induction of type 2diabetes (T2DM) was done by intraperitoneal infusion of streptozotocin. The progressive aerobic training protocol entailed 4 weeks, 5 sessions per week running on treadmill at velocity of 20 m/min for 60 min. The gene expression of TNF-α, VEGF and HIF-1α were measured by real time &amp; PCR. One way analysis of variance and bonferroni test were applied to analysis the data. The significant level was set at P-value&lt; 0.05.&#x0D; Results: The results indicated that the aerobic training induced significant decrease in TNF-α mRNA (P-value&lt; 0.001) and significant increase in HIF-1α mRNA (P-value&lt; 0.001) and VEGF mRNA (P-value&lt; 0.001) compared to diabetic control group.&#x0D; Conclusion: It appears that aerobic training with reduction of TNF-α issues an affirmative effect on angiogenesis, as a result, it improves diabetic cardiac.

MicroRNAs miR-1 and miR-206 Regulate Monocarboxylate Transporter-4 and Vascular Endothelial Growth Factor Gene Expression in Colorectal Cancer

Background: Colorectal cancer (CRC) is currently the third most common cancer type in males and the second most occurring in females. The role of microRNA (miRNA) in the development of colorectal cancer is not fully elucidated. Therefore, understanding the mechanistic interaction between miRNA and their target oncogenes may hold great importance as a possible target for interventional anticancer therapy.&#x0D; Aims: To identify miRNAs that are part of the regulating pathway of Monocarboxylate Transporter-4 (MCT4) and Vascular Endothelial Growth Factor (VEGF) oncogenes.&#x0D; Study Design: We used publicly available prediction tools (e.g. TargetScan, MicroCosm, PicTar, and DIANA-microT-CDS) to identify the possible miRNA that target the two oncogenes.&#x0D; Methodology: We used the GeneMania database to visualize the network and verify gene names and remove ambiguity and duplications. Furthermore, we used miRTarBase database to identify experimentally validated targets which we used to further confirm miRNA-oncogene relationships. Finally, we utilized miR-Mfold web-tool to further visualize the circular structures and the simulated miR-1 and miR-206 targeting arrangements.&#x0D; Results: We found two putative miRNA (miR-1 and miR-206) that may downregulate MCT4 coded by SLC16A3 gene and VEGF which is coded by VEGF gene. We found relationships between the validated target genes of miR-1 and miR-206 through GeneMania which we extracted from the literature. And we elucidated the proposed structure of these two miRNAs through miR-Mfold web-tool.&#x0D; Conclusion: Our results elucidated a novel regulation pathway in CRC cells and may suggest a potential therapeutic approach for CRC therapy. MiR-1 and miR-206 may help cells go to apoptosis and inhibit the angiogenesis of colorectal cancer cells by down-regulation of MCT4 and VEGF proteins in tumor tissues.

Characterization of a Thermo-Sensitive Injectable Hydrogel as an Iloprost Delivery System for Dental Use

Iloprost increases the expression of angiogenic factors and increases dental pulp flow, suggesting the potential of iloprost as a biomolecule to promote dental pulp regeneration. However, the methods to clinically deliver iloprost into the limited root canal area of a tooth and control its release are limited. The purpose of this study was to prepare a thermo-sensitive injectable hydrogel from pluronic F127 (PF127) for delivering iloprost to induce dental pulp regeneration. The PF127 hydrogels were fabricated using thermal crosslinking. The maximum cumulative release iloprost from the hydrogel at 25°C was 60%. No significant cytotoxicity or morphological changes were observed in human dental pulp cells (HDPCs) at any of the PF127 gel concentrations of the iloprost carrier. Moreover, the effect of the 20%wt PF127 gels containing iloprost on the expression of VEGF in HDPCs increased vascular endothelial growth factor (VEGF) gene expression at 72 h. The thermo-sensitive hydrogel at 20%wt PF127 containing iloprost could be used for prolonged drug release in dental applications.

Angiogenesis is Differentially Modulated by Anthocyanin and Phenolic Acid Extracts from Wild Blueberry (V. angustifolium) Through PI3K Pathway.

Wild blueberries rank very high in anthocyanins (ACNs) and phenolic acids (PA) among other berries. Previous work from our group has documented their beneficial effects. In this study, human umbilical vein endothelial cells were used, and ACNs, PAs, and their combination (ACNs:PAs) at concentrations of 0.002, 8, 15, 60, and 120 μg/mL were tested for endothelial tube formation. Treatment with ACNs decreased, while treatment with PAs and ACNs:PAs increased overall endothelial cell tube formation compared to control. Endothelial cells exposed to ACNs downregulated gene expression of AKT1 and endothelial nitric oxide synthase (eNOS), while PAs upregulated AKT1 and vascular endothelial growth factor (VEGF) gene expression. Combination of ACNs:PAs decreased gene expression of AKT1 and eNOS, while protein levels of AKT1 increased. In summary, based on the type of wild blueberry extract, angiogenesis is differentially modulated and is concentration dependent. Further experiments will delineate the mechanism(s) of the differential action of the aforementioned extracts on angiogenesis.

In vitro effects of photobiomodulation applied to gingival fibroblasts cultured on titanium and zirconia surfaces and exposed to LPS from Escherichia coli.

Photobiomodulation (PBM) therapy is used to stimulate cell proliferation and metabolism, as well as reduce inflammatory cytokine synthesis, which plays a main role in the long-term stability of implants. This study assessed the response of gingival fibroblasts cultured on titanium (Ti) and zirconia (ZrO2), submitted to PBM and exposed to lipopolysaccharide (LPS). Cells seeded on Ti and ZrO2 were irradiated (InGaAsP; 780nm, 25mW) 3 times, using 0.5, 1.5, and 3.0J/cm2 doses, and exposed to Escherichia coli LPS (1μg/mL). After 24h, cell viability (alamarBlue, n = 8), interleukin 6 (IL-6) and 8 (IL-8) synthesis (ELISA, n = 6), and IL-6 and vascular endothelial growth factor (VEGF) gene expression (qPCR, n = 5) were assessed and statistically analyzed (one-way ANOVA, α = 0.05). Cell morphology was evaluated by fluorescence microscopy. Increased cell viability occurred in all groups cultured on Ti compared with that of the control, except for cells exposed to LPS. Fibroblasts cultured on ZrO2 and LPS-exposed exhibited reduced viability. PBM at 3.0J/cm2 and 1.5J/cm2 downregulated the IL-6 synthesis by fibroblasts seeded on Ti and ZrO2, as well as IL-8 synthesis by cells seeded on ZrO2. Fibroblasts seeded on both surfaces and LPS-exposed showed increased IL-6 gene expression; however, this activity was downregulated when fibroblasts were irradiated at 3.0J/cm2. Enhanced VEGF gene expression by cells seeded on Ti and laser-irradiated (3.0J/cm2). Distinct patterns of cytoskeleton occurred in laser-irradiated cells exposed to LPS. Specific parameters of PBM can biomodulate the inflammatory response of fibroblasts seeded on Ti or ZrO2 and exposed to LPS.