As a novel endogenous anti-angiogenic molecule, vasohibin 1 (VASH1) is not only expressed in tumor stroma, but also in tumor tissue. Moreover, studies have shown that VASH1 may be a prognostic marker in colorectal cancer (CRC). Knockdown of VASH1 enhanced transforming growth factor-β1 (TGF-β1)/Smad3 pathway activity and type I/III collagen production. Our previous findings suggest that ELL-associated factor 2 (EAF2) may play a tumor suppressor and protective role in the development and progression of CRC by regulating signal transducer and activator of transcription 3 (STAT3)/TGF-β1 signaling pathway. However, the functional role and mechanism of VASH1-mediated TGF-β1 related pathway in CRC has not been elucidated.

To investigate the expression of VASH1 in CRC and its correlation with the expression of EAF2. Furthermore, we studied the functional role and mechanism of VASH1 involved in the regulation and protection of EAF2 in CRC cells in vitro.

We collected colorectal adenocarcinoma and corresponding adjacent tissues to investigate the clinical expression of EAF2 protein and VASH1 protein in patients with advanced CRC. Following, we investigated the effect and mechanism of EAF2 and VASH1 on the invasion, migration and angiogenesis of CRC cells in vitro using plasmid transfection.

Our findings indicated that EAF2 was down-regulated and VASH1 was up-regulated in advanced CRC tissue compared to normal colorectal tissue. Kaplan-Meier survival analysis showed that the higher EAF2 Level group and the lower VASH1 Level group had a higher survival rate. Overexpression of EAF2 might inhibit the activity of STAT3/TGF-β1 pathway by up-regulating the expression of VASH1, and then weaken the invasion, migration and angiogenesis of CRC cells.

This study suggests that EAF2 and VASH1 may serve as new diagnostic and prognostic markers for CRC, and provide a clinical basis for exploring new biomarkers for CRC. This study complements the mechanism of EAF2 in CRC cells, enriches the role and mechanism of CRC cell-derived VASH1, and provides a new possible subtype of CRC as a therapeutic target of STAT3/TGF-β1 pathway.

• ELL-associated factor 2
• Vasohibin 1
• Transforming growth factor-β1
• Signal transducer and activator of transcription 3
• Colorectal cancer
• Angiogenesis

• Humans
• Transforming Growth Factor beta1
• Transcription Factors/genetics
• Colorectal Neoplasms/pathology
• Signal Transduction
• Transfection
• Cell Line, Tumor
• Transcriptional Elongation Factors
• Cell Cycle Proteins/genetics
• Cell Cycle Proteins/metabolism

Objective: Trimethylamine-N-oxide (TMAO) was found to play crucial roles in vascular endothelial function. However, the exact molecular mechanisms are not yet entirely clear. Recently, we found that exosomes (Exos) isolated from TMAO-treated hepatocytes (TMAO-Exos) contained a distinctive profile of miRNAs compared to those from the TMAO-free group (Control-Exos). Furthermore, TMAO-Exos could notably promote inflammation, damage vascular endothelial cells (VECs), and impair endothelium-dependent vasodilation. This study aimed to further evaluate the effects of TMAO-Exos on VECs and explore the underlying mechanisms.

Methods: Exos were isolated from the hepatocyte culture supernatant with or without TMAO, using differential centrifugation. Then, VECs were treated with these Exos for 48 h and subjected to RNA-sequencing for detecting the changes of alternative polyadenylation (APA) and mRNA. After validation by qPCR and western blotting, the recombinant viruses were used to mediate the overexpression of C-X-C motif chemokine receptor 4 (CXCR4). The in vitro VEC function was evaluated by cell migration and tube formation, and in vivo angiogenesis was investigated in hindlimb ischemia models.

Results: Exos released from hepatocytes were differentially regulated by TMAO; both could be taken up by VECs; and furthermore, TMAO-Exos significantly reduced cell migration and tube formation in vitro and impaired perfusion recovery and angiogenesis after hindlimb ischemia, by down-regulating the CXCR4 expression. However, TMAO-Exos failed to regulate the splicing events, at least in this experimental setting, which suggested that TMAO-Exos may affect CXCR4 expression via an APA-independent manner.

Conclusions: Our findings revealed a novel indirect mechanism by which TMAO impaired endothelial function through stimulating hepatocytes to produce Exos that possessed distinctive activity. The crosstalk between the liver and vascular endothelial mediated by these Exos may offer a new target for restraining the harmful effects induced by TMAO.

• C-X-C motif chemokine receptor 4
• alternative polyadenylation
• angiogenesis
• endothelial function
• hepatocyte-derived exosomes
• trimethylamine-N-oxide

Hypoxia, a major inducer of angiogenesis, triggers major changes in gene expression at the transcriptional level. Furthermore, under hypoxia, global protein synthesis is blocked while internal ribosome entry sites (IRES) allow specific mRNAs to be translated. Here, we report the transcriptome and translatome signatures of (lymph)angiogenic genes in hypoxic HL-1 mouse cardiomyocytes: most genes are induced at the translatome level, including all IRES-containing mRNAs. Our data reveal activation of (lymph)angiogenic factor mRNA IRESs in early hypoxia. We identify vasohibin1 (VASH1) as an IRES trans-acting factor (ITAF) that is able to bind RNA and to activate the FGF1 IRES in hypoxia, but which tends to inhibit several IRESs in normoxia. VASH1 depletion has a wide impact on the translatome of (lymph)angiogenesis genes, suggesting that this protein can regulate translation positively or negatively in early hypoxia. Translational control thus appears as a pivotal process triggering new vessel formation in ischemic heart.

• IRES
• angiogenesis
• cardiomyocyte
• cell biology
• chromosomes
• gene expression
• hypoxia
• mouse
• translational control
• vasohibin

• FGFR
• HIF-1
• NRP
• VEGF
• VEGFR
• alternative splicing
• angiogenesis
• angiopoietin
• vasohibin

• Alternative Splicing/genetics
• Alternative Splicing/physiology
• Angiopoietins/genetics
• Angiopoietins/metabolism
• Animals
• Humans
• Neovascularization, Pathologic/genetics
• Neovascularization, Pathologic/metabolism
• Receptors, Vascular Endothelial Growth Factor/genetics
• Receptors, Vascular Endothelial Growth Factor/metabolism

• PG/15/53/31371 British Heart Foundation
• 15/53/31371 British Heart Foundation
• 17/0005668 Diabetes UK

• Prostate cancer
• Robo 4
• Slit 2
• cancer aggressiveness
• tumor recurrence

• TAp73
• angiogenesis
• immunohistochemistry
• lung adenocarcinoma
• p53
• vasohibin-1

Vasohibin-1 is an intrinsic angiogenesis inhibitor, and is expressed in endothelial cells via induction by pro-angiogenesis factors. It is known to inhibit several processes of angiogenesis, with different mechanisms from extrinsic angiogenesis inhibitors. Vasohibin-2 is mainly expressed by mononuclear cells which have been mobilized from bone marrow. It not only promotes angiogenesis, but also modulates the releases of FGF-2 and VEGF, which are the two major inducers for vasohibin1. Hypoxic environment induces the expression of hypoxia-inducible Factor 1α with a result of VEGF release nearly in all tumor cell lines and tissues. However, it has been observed that hypoxia reduces the inducible effects of VEGF on vasohibin, which indicates that a complicated mechanism exists in the angiogenesis. Vasohibin and its family members play important roles in both the physiological and pathological procedures, in contrary but complementary patterns. Furthermore, human aortic smooth muscle cells and fibroblast have also been detected to express vasohibin on a moderate to weak scale range. Recently, the results of an increasing number of studies in vivo have shown that vasohibin can also be detected in several cancers, and is associated with micro-vessel densities, histology grades, invasions, poor clinical features, metastasis, and dissemination in abdominal cavities, as well as EMT. In more recent reports, it has been confirmed that, along with being angiogenesis regulators, a variety of other roles have been associated with this family. The focus of this study was the upstream regulatory mechanisms of vasohibin expressions, and their role in regard to the downstream target proteins of vasohibin, especially in carcinoma. Vasohibin is considered to be an original angiogenesis inhibitor, and has a much broader significance in pathological processes. It can be taken as an independent prognostic factor, as well as a potential strategy for cancer therapy programs.

• angiogenesis
• angiogenesis inhibitor
• carcinogenesis
• growth and differentiation regulating factors
• natural anticancer compounds
• prognostic marker
• tumors

• angiogenesis
• immunohistochemistry
• prognosis
• urinary bladder neoplasms
• vasohibin-1

• 肺鳞状细胞癌
• vasohibin-1
• macc1
• 免疫组织化学
• 预后
• lung squamous cell carcinoma
• immunohistochemistry
• prognosis

• VASH1
• angiogenesis
• colon cancer
• metastasis
• tumor suppressor

Previously we isolated migracin A from a Streptomyces culture filtrate as an inhibitor of cancer cell migration. In the present research, we found that migracin A inhibited migration and invasion of ovarian clear cell carcinoma ES-2 cells. In the course of our mechanistic study, migracin A was shown to enhance vasohibin-1 expression in an angiogenesis array. We also confirmed that it increased the mRNA expression of this protein. Moreover, overexpression of vasohibin-1 lowered the migration but not the invasion of ES-2 cells. Then, we looked for another target protein employing a motility array, and found that migracin A lowered the IGF-1 expression. Knockdown of IGF-1 by siRNA decreased the migration and invasion of ES-2 cells. Migracin A also decreased Akt phosphorylation involved in the downstream signaling. Crosstalk analysis indicated that overexpression of vasohibin-1 decreased the IGF-1 expression. On the other hand, it showed no direct anticancer activity in terms of the ES-2 growth in agar. Migracin A inhibited the migration and IGF-1 expression in not only ES-2 but also another ovarian clear cell carcinoma JHOC-5 cells. In addition, it also inhibited capillary tube formation of human umbilical vein endothelial cells. Since its cytotoxicity is very low, migracin A may be a candidate for an anti-metastasis agent not exhibiting prominent toxicity.

• Adenocarcinoma, Clear Cell/genetics
• Adenocarcinoma, Clear Cell/metabolism
• Benzaldehydes/pharmacology
• Cell Cycle Proteins/genetics
• Cell Cycle Proteins/metabolism
• Cell Line, Tumor
• Cell Movement/drug effects
• Cell Movement/genetics
• Female
• Gene Expression
• Humans
• Insulin-Like Growth Factor I/antagonists & inhibitors
• Insulin-Like Growth Factor I/genetics
• Insulin-Like Growth Factor I/metabolism
• Neovascularization, Pathologic
• Ovarian Neoplasms/genetics
• Ovarian Neoplasms/metabolism
• Phosphoinositide-3 Kinase Inhibitors
• Protein Kinase Inhibitors/pharmacology
• Proteome
• Proteomics/methods
• Proto-Oncogene Proteins c-akt/antagonists & inhibitors
• Receptor, IGF Type 1/metabolism
• Signal Transduction/drug effects
• Tumor Stem Cell Assay

• alternative splicing
• pre-mrna
• gene function

• Alternative Splicing
• Animals
• Evolution, Molecular
• Exons
• Gene Expression
• Humans
• Models, Genetic
• Protein Isoforms/genetics
• Protein Isoforms/metabolism
• RNA, Messenger/genetics
• RNA, Messenger/metabolism
• Transcription Factors/metabolism

• R01 GM083187 NIGMS NIH HHS

• antiangiogenesis
• chorioallantoic membrane
• tumor progression

Angiogenesis is regulated by the local balance between angiogenesis stimulators and inhibitors. A number of endogenous angiogenesis inhibitors have been found in the body. The origin of these inhibitors is mostly extrinsic to the vasculature. Recently, however, vascular endothelial cells themselves have been found to produce angiogenesis inhibitors including vasohibin-1. These intrinsic inhibitors are thought to regulate angiogenesis by an auto-regulatory or negative-feedback mechanism. This review will focus on vasohibin-1 produced by vascular endothelial cells and on its homologue, vasohibin-2.

• bone marrow
• endothelial cell
• mononuclear cell
• negative feedback