miR-29c-3p represses the angiogenesis of esophageal squamous cell carcinoma by targeting SERPINH1 to regulate the Wnt signaling pathway

Purpose: Esophageal squamous cell carcinoma (ESCC) is characterized by early metastasis and late diagnosis. miR-29c-3p is confirmed to repress angiogenesis in multiple tumor types. Yet, the functions of miR-29c-3p in the mechanism of ESCC angiogenesis, which were not sufficiently explored previously, were exactly what we investigated here at the molecular level. Methods: The mRNA level of miR-29c-3p and Serpin peptidase inhibitor clade H member 1 (SERPINH1) in ESCC tissues were assessed via bioinformatics analysis. Thereafter, miR-29c-3p and SERPINH1 (HSP47) mRNA level in ESCC cell lines was evaluated via quantitative real-time polymerase chain reaction. The effects of abnormal miR-29c-3p and SERPINH1 expression on ESCC cell viability, proliferation, migration, invasion, and HUVEC angiogenesis were examined via CCK8, colony formation, transwell, and angiogenesis assays, respectively. The protein levels of SERPINH1, vascular endothelial growth factor-A (VEGFA), Wnt-1, ?-catenin, and p-?-catenin were evaluated via Western blot. Expression of VEGFA secreted by ESCC cells was measured via enzyme-linked immunosorbent assay. Treatment with the Wnt activator BML-284 further revealed the way miR-29c-3p mediated the Wnt signaling pathway and its effects on angiogenesis. Results: Herein, we revealed a decrease of miR-29c-3p expression in ESCC tissues and cells, while the overexpressed miR-29c-3p could remarkably suppress ESCC cell progression, as well as HUVEC angiogenesis. Meanwhile, overexpressed miR-29c-3p notably downregulated VEGFA and repressed the Wnt signaling pathway. Treatment with the Wnt activator BML-284 could reverse the inhibition of HUVEC angiogenesis caused by miR-29c-3p. SERPINH1 was a downstream target of miR-29c-3p. SERPINH1 knockdown suppressed the malignant phenotypes of ESCC cells and impeded the Wnt signaling activation, while such suppression was reversed through miR-29c-3p inhibitor. Conclusions: We confirmed the mechanism that miR-29c-3p targeted SERPINH1, thus regulating angiogenesis in ESCC through the Wnt signaling pathway. It improves the understanding of angiogenesis in ESCC and offers new ideas for the research of ESCC treatment strategies in the future.

IARS2 regulates proliferation, migration, and angiogenesis of human umbilical vein endothelial cells

SUMMARY OBJECTIVE: In this study, we aimed at investigating the role of isoleucyl-tRNA synthetase in the growth, migration, and angiogenesis of human umbilical vein endothelial cells and the underlying molecular mechanism. METHODS: To assess the role of isoleucyl-tRNA synthetase, we silenced isoleucyl-tRNA synthetase in human umbilical vein endothelial cells using lentiviral 2 specific short hairpin RNAs (short hairpin RNAs 1 and 2) and examined silencing efficiency using real time quantitative polymerase chain reaction and western blot analyses. Short hairpin RNAs 1-isoleucyl-tRNA synthetase had greater knockdown efficiency, it was used in the entire downstream analysis. Short hairpin RNAs 1- isoleucyl-tRNA synthetase silencing effects on cell proliferation, cell colony generation, cell migration, as well as angiogenesis were assessed using cell counting kit-8, colony development, cell migration, and angiogenesis tube formation assays, respectively. RESULTS: Compared to the control group, anti-isoleucyl-tRNA synthetase short hairpin RNAs significantly silenced isoleucyl-tRNA synthetase expression in human umbilical vein endothelial cells, and suppressed their proliferation, migration, and angiogenic capacity. To characterize the underlying mechanism, western blot analyses showed that isoleucyl-tRNA synthetase knockdown suppressed phosphorylation of extracellular-regulated kinase ½ and protein-serine- threonine kinase, as well as expression of vascular endothelial growth factor, GSK-3β, and β-catenin. CONCLUSIONS: We have shown, for the first time, the critical role of isoleucyl-tRNA synthetase in human umbilical vein endothelial cells. Our data show that isoleucyl-tRNA synthetase knockdown suppresses human umbilical vein endothelial cell proliferation, migration, and angiogenesis. We have also shown that isoleucyl-tRNA synthetase knockdown suppresses phosphorylation of extracellular-regulated kinase ½ and protein-serine- threonine kinase, as well as expression of vascular endothelial growth factor, GSK-3β, and β-catenin. Together, these data highlight isoleucyl-tRNA synthetase as a potential antitumor anti-angiogenic target.

Correlation of Angiogenic Factor AGGF1 Expression With Prognosis of Patients With Colon Cancer / 胃肠病学

Background:Angiogenesis plays a critical role in tumor growth and metastasis. AGGF1,a new member of angiogenic factors,has been shown to be aberrantly expressed in a variety of malignant tumors. Aims:To investigate the correlation of AGGF1 expression with prognosis of patients with colon cancer. Methods:Eighty colon cancer patients undergoing surgical operation from Jan. 2010 to Dec. 2012 at Xi’an First Hospital were recruited. Immunohistochemistry was performed to evaluate the expression of AGGF1 in cancerous and paired paracancerous tissues. The correlations of AGGF1 expression in cancerous tissue with clinicopathological characteristics and prognosis were analyzed. Cox regression model was used to identify the prognostic factors of colon cancer. Results:AGGF1 expression was significantly higher in colon cancer tissue than in paracancerous tissue (67. 5% vs. 32. 5%,P＜0. 05),and was correlated with tumor differentiation,lymph node metastasis,vascular/lymphatic infiltration and TNM stage (P＜0. 05). Kaplan-Meier survival analysis showed that high AGGF1 expression was correlated with decrease of survival rate in colon cancer patients (P ＜0. 05). Univariate and multivariate analyses identified AGGF1 as a prognostic factor of patients with colon cancer (OR＝2. 09,95% CI:1. 67-4. 45,P＝0. 011;OR ＝1. 98,95% CI:1. 72-4. 59,P ＝0. 000 ). Conclusions:Angiogenic factor AGGF1 is up-regulated and correlated with tumor metastasis and unfavorable prognosis in patients with colon cancer. It might be a potential prognostic indicator and deserves further study.

Gastric Cancer and Angiogenesis: Is VEGF a Useful Biomarker to Assess Progression and Remission?

Gastric cancer (GC) has high mortality owing to its aggressive nature. Tumor angiogenesis plays an essential role in the growth, invasion, and metastatic spread of GC. The aim of this work was to review the angiogenic biomarkers related to the behavior of GC, documented in the literature. A search of the PubMed database was conducted with the MeSH terms: “Stomach neoplasms/blood [MeSH] or stomach neoplasms/blood supply [MeSH] and angiogenic proteins/blood [Major]”. A total of 30 articles were initially collected, and 4 were subsequently excluded. Among the 26 articles collected, 16 examined the role of vascular endothelial growth factor (VEGF), 4 studied endostatin, 3 investigated angiopoietin (Ang)-2, 2 studied the Ang-like protein 2 (ANGTPL2), and 1 each examined interleukin (IL)-12, IL-8, and hypoxia inducible factor. Regarding VEGF, 6 articles concluded that the protein was related to lymph node metastasis or distant metastases. Five articles concluded that VEGF levels were elevated in the presence of GC and decreased following tumor regression, suggesting that VEGF levels could be a predictor of recurrence. Four articles concluded that high VEGF levels were correlated with poor prognosis and lower survival rates. Ang-2 and ANGTPL2 were elevated in GC and associated with more aggressive disease. Endostatin was associated with intestinal GC. VEGF is the most extensively studied angiogenic factor. It is associated with the presence of neoplastic disease and lymph node metastasis. It appears to be a good biomarker for disease progression and remission, but not for diagnosis. The data regarding other biomarkers are inconclusive.

Genetic Contributions to Childhood Obesity: Association of Candidate Gene Polymorphisms and Overweight/Obesity in Korean Preschool Children

This study was aimed to investigate the association of candidate gene polymorphisms and obesity or overweight in young Korean children. A total of 190 Korean preschool children (96 control, 48 overweight, and 46 obese children) were genotyped for the angiotensin converting enzyme (ACE) insertion (I)/deletion (D), angiotensin II type 2 receptor (AT2) C3123A, transforming growth factor (TGF)-β1 T869C, vascular endothelial growth factor (VEGF) T460C, and tumor necrosis factor (TNF)-α G308A polymorphisms. No differences were found among the groups with respect to age, sex, birth weight, blood pressure levels, and serum concentrations of glucose and total cholesterol. Obese children showed a higher incidence of ACE DD genotype and D allelic frequency compared to the controls (odds ratio [OR], 2.7, 95% confidence interval [CI], 1.01–7.21; OR, 2.5, 95% CI, 1.49–4.19; all P < 0.05). The frequency of TC genotype and C allele in the TGF-β1 T869C polymorphism (OR, 2.08, 95% CI, 1.01–4.27; OR, 1.93, 95% CI, 1.15–3.21) and that in the VEGF T460C polymorphism (OR, 2.5, 95% CI, 1.19–5.28; OR, 2.15, 95% CI, 1.26–3.68) was also higher in obese children than in control subjects (all P < 0.05). Overweight children exhibited a higher frequency of the A allele in the AT2 C3123A polymorphism compared to the controls (OR, 1.72, 95% CI, 1.03–2.88, P < 0.05). There were no differences in the TNF-α G308A polymorphism among the groups. The ACE I/D, AT2 C3123A, TGF-β1 T869C, and VEGF T460C polymorphisms can affect susceptibility to obesity or overweight in Korean children.

Effect of propranolol on the expression of growth factors and apoptotic factors related to infantile hemangiomas / 解放军医学杂志

Objective To investigate the effects of propranolol on the expression of growth factors and apoptotic factors related to infantile hemangiomas, and explore the underlying mechanisms for treatment of hemangiomas by propranolol. Methods Oral propranolol was administered to 39 patients ≤3 months with hemangiomas during the proliferative phase, and they were in accordance with the inclusion criteria of the treatment. Patients who had contraindications to treatment were excluded, and informed consent from their family members was obtained. Biopsy of the lesion under local anesthesia was done before medication. The oral dose of propranolol was 1mg/kg per 12 hours. After 8 weeks of treatment, the hemangioma was resected. The specimens taken before and after treatment were scrutinized for changes in tissue structure and cell form after HE staining. The expressions of the proteins AD RB1, ADRB2, ERK, Akt, NF-kB, VEGFA, Cyclin D1, Cyclin E1, Ki-67, Bax, Bcl-2, caspase-8, Fas, FasL, caspase-9, and caspase-3 in the focal tissue were determined with immunohistochemistry and real-time fluorescence quantitative RT-PCR. At the same time, the microvessel density (MVD) value, Ki-67 index and apoptosis index before and after treatment were also determined and compared using CD34, Ki-67 and TUNEL staining technique, respectively. Meanwhile, the mRNA expressions of these factors were assessed with quantitative real-time RT-PCR. ResultsAfter 8 weeks of treatment, all the protein content and mRNA expressions of ADRB1, ADRB2, ERK, Akt, NF-κB, VEGFA, Cyclin D1, Cyclin E1, Ki-67, Bcl-2, and MVD value and Ki-67 index were decreased significantly (P<0.01), while the protein and mRNA expressions of Bax, caspase-3, caspase-9, and apoptosis index were increased significantly (P<0.01). However, there was no obvious change in the expressions of Fas, FasL and caspase-8 after treatment. ConclusionPropranolol can inhibit proliferation and promot intrinsic apoptotic pathway of hemangioma through regulating the MAPKs and PI3K-Akt pathways, but it shows no influence on the extrinsic apoptotic pathway.

Genetics and pathophysiological mechanisms of moyamoya disease / 国际脑血管病杂志

Moyamoya disease (MMD) is a chronic and progressive cerebrovascular disease which is characterized by the bilateral internal carotid artery ends and (or) stenosis or occlusion of anterior cerebral artery and middle cerebral artery initial segments,compensatory proliferation of small blood vessels in the skull base and formation of abnormal vascular network.Its etiology and pathogenesis remains unclear.The present studies speculate that MMD may be a polygenic disease,inflammation,immune response,abnormal cytokine secretion,endothelial progenitor cell change and nitric oxide level change are associated with the occurrence and development of MMD.This article reviews the advances in research on the genetics and pathophysiological mechanism of MMD.

Molecular mechanisms of angiogenesis after cerebral ischemia / 国际脑血管病杂志

Angiogenesis is an important self-repair and remodeling mechanism after cerebral ischemia. It plays pivotal roles in promoting local blood circulation, protecting neurons and improving neurological function after cerebral ischemia. A variety of molecules and signal transduction pathways involved in the regulation of the process, including angiogenin, growth factors, matrix metalloproteinases, tissue kallikrein and anti-angiogenic proteins. This article reviews the roles of these molecules and signal transduction pathways in angiogenesis after cerebral ischemia.

Aspectos celulares, teciduais e moleculares da osteogênese ectópica e ortotópica induzida pela matriz alogênica óssea e dentinária / Cellular, tissue and molecular aspects of the ectopic and orthotopic osteogenese induced by bone and dentine allogenic matrix

O objetivo do atual trabalho, foi correlacionar os eventos celulares e teciduais com a expressão das proteínas VEGF, BMP-7, RANKL e OPG durante a osteogênese ectópica e ortotópica, induzida pela matriz óssea (MO) e dentinária (MD) alogênica. Matrizes alogênicas desmineralizada em HCl a 0,6N, obtidas de fêmur e incisivo de ratos, fori implantada entre as fáscias musculares da coxa e em defeito trans-ósseo de 8mm de diâmetro nos ossos parietais. As análises radiográfica e histomorfométrica da neoformação óssea e, a imunohistoquímica e o western blotting para as proteínas VEGF, BMP, RANKL e OPG, mostraram que: a) o volume da região do enxerto nos sítios ortotópicos reduziu 19% em 42 dias; b) em ambos tipos de enxerto e locais de implantação, ocorreu formação de tecido cartilaginoso e ósseo; c) nos sítios intramusculares, a reabsorção da matriz alogênica e a remodelação do tecido cartilaginoso, ósseo e medular foi mais acelerado, em relação a implantação ortotópico; d) o aumento na densidade de volume dos vasos sanguíneos e no número de osteoblastos/osteócitos e osteoclastos ocorreu simultaneamente e estava associado à maior reabsorção da matriz alogênica e à formação do tecido medular (hematopoiético); e) as proteínas VEGF, BMP-7, RANKL, OPG foram expressas em condrócitos, osteoblastos ativos, osteócitos recém aprisionados na matriz e em células estromais próximas aos osteoblastos ou às áreas da matriz alogênica reabsorvida; e f) a expressão das proteínas VEGF, BMP-7, RANKL e OPG foi maior no grupo MO. O pico de expressão dessas proteínas ocorreu nos períodos de 14 aos 21 dias no grupo da MO e 21 e 28 dias no grupo da MD.

Concluímos que, a capacidade osteoindutora da matriz alogênica desmineralizada está relacionado a origem da matriz e ao sítio de implantação e que, as proteínas VEGF, BMP-7, RANKL e OPG estão associadas a maior reabsorção da matriz implantada, promovendo uma rápida e contínua liberação dos morfógenos contidos em seu interior que, induzem temporal e espacialmente a formação óssea/medular.

The aim of the present work was to correlate the cellular and tissue events with the expression of VEGF, BMP-7, RANKL and OPG during ectopic and orthotopic osteogenesis, induced by bone and dentin allogeneic matrix. Allogenic matrices obtained from femur and incisor of rats and demineralized in 0.6 N HCl were implanted into a intramuscular pocket and a 8mm-diameter bone defect in the skull. The radiographic and histomorphometric analysis of new bone formation, and immunohistochemistry and western blotting for VEGF, BMP, RANKL and OPG proteins, showed that: a) the total volume of the graft region in orthotopic site decreased 19% at 42 days b) in both graft types and implantation sites occurred formation of cartilaginous and bone tissues, c) in intramuscular sites, the resorption of allogenic matrix and remodeling of the new formed cartilage and bone were faster, in relation to orthotopic implantation sites; d) the increase in the volume density of blood vessels and in the number of osteoblasts/osteocytes and osteoclasts occurred simultaneously and was associated with greater reabsorption of the allogenic matrix and hematopoietic bone marrow formation; e) VEGF, BMP-7, RANKL, OPG proteins were expressed in chondrocytes, active osteoblasts, newly osteocytes confined and stromal cells located near the osteoblasts or in the surface of the reabsorbed matrix; and f) the VEGF, BMP-7, RANKL and OPG expression was higher in MO grafts than in the MD. The peak of expression of these proteins each occurred at 14 and 21 days in MO and 21 and 28 days in MD. We concluded that, the osteoinductive capacity of allogeneic demineralized matrix is related to matrix origin and implantation site and that the VEGF, BMP-7, RANKL and OPG proteins are associated with greater reabsorption of the implanted matrix, promoting rapid and continuous matrix-release morphogens that induces spatially and temporally the bone and bone marrow formation.

Hypoxia and angiogenesis: regulation of hypoxia-inducible factors via novel binding factors

The mechanisms that regulate angiogenesis in hypoxia or hypoxic microenvironment are modulated by several pro- and antiangiogenic factors. Hypoxia-inducible factors (HIFs) have been established as the basic and major inducers of angiogenesis, but understanding the role of interacting proteins is becoming increasingly important to elucidate the angiogenic processes of a hypoxic response. In particular, with regard to wound healing and the novel therapies for vascular disorders such as ischemic brain and heart attack, it is essential to gain insights in the formation and regulation of HIF transcriptional machineries related to angiogenesis. Further, identification of alternative ways of inhibiting tumor growth by disrupting the growth-triggering mechanisms of increasing vascular supply via angiogenesis depends on the knowledge of how tumor cells develop their own vasculature. Here, we review our findings on the interactions of basic HIFs, HIF-1alpha and HIF-2alpha, with their regulatory binding proteins, histone deacetylase 7 (HDAC7) and translation initiation factor 6 (Int6), respectively. The present results and discussion revealed new regulatory interactions of HIF-related mechanisms.

Expression of endothelial nitric oxide synthase correlates with the angiogenic phenotypes and poor prognosis of human gastric cancer / 肿瘤

Objective: Angiogenesis is a critical step in tumor growth and is regulated by multiple molecular pathways. We evaluted the expression of nitric oxide synthase (NOS) III and its relationship with the angiogenic phenotypes and expressions of the transcription factor Sp1, as well as their effect on survival rate of patients with gastric cancer. Methods: The expression levels of NOS III and Sp1, and tumor microvessel density (MVD) status in 86 specimens of resected gastric cancer tissues were determined by immunohistochemistry. Results: NOS III protein expression was significantly higher in both primary tumors and lymph node metastases than in normal gastric mucosa. In primary tumors, NOS III expression had close correlation with Sp1 expression (P=0.001) and MVD status (P=0.001). Patients with positive Sp1 expression were more likely to have strong NOS III expression (15 times) and a high MVD (7 times) than those with negative Sp1 expression. Univariate survival analysis showed that strong NOS III expression, strong Sp1 expression, and a high MVD were associated with poor prognosis. In a Cox proportional hazards model, only strong NOS III and Sp1 expression as well as advanced disease stage were independent factors for prognosis of patients. Conclusion: Our results indicate that the expression of NOS III may play an important role in tumor angiogenesis and infiltration of gastric cancer.