Formation Of Tubular Structure Research Articles

Effects of adipose-derived stem cell released exosomes on proliferation, migration, and tube-like differentiation of human umbilical vein endothelial cells

To explore the effects of adipose-derived stem cell released exosomes (ADSC-Exos) on the proliferation, migration, and tube-like differentiation of human umbilical vein endothelial cells (HUVECs). Adipose tissue voluntarily donated by liposuction patients was obtained. The ADSCs were harvested by enzyme digestion and identified by flow cytometry and adipogenic induction. The ADSC-Exos were extracted from the supernatant of the 3rd generation ADSCs and the morphology was observed by transmission electron microscopy. The surface proteins (Alix and CD63) were detected by Western blot. The nanoparticle tracking analyzer NanoSight was used to analyze the size distribution of ADSC-Exos. After co-culture of PKH26 fluorescently labeled ADSC-Exos with HUVECs, confocal microscopy had been used to observe whether ADSC-Exos could absorbed by HUVECs. ADSC-Exos and HUVECs were co-cultured for 1, 2, 3, 4, and 5 days. The effect of ADSC-Exos on the proliferation of HUVECs was detected by cell counting kit 8 (CCK-8) assay. The expression of VEGF protein in the supernatant of HUVECs with or without ADSC-Exos had been detected by ELISA after 12 hours. Transwell migration assay was used to detect the effect of ADSC-Exos on the migration ability of HUVECs. The effect of ADSC-Exos on the tubular structure formation of HUVECs was observed by Matrigel experiments in vitro. The formation of subcutaneous tubular structure in vivo was observed in BALB/c male nude mice via the injection of HUVECs and Matrigel with or without ADSC-Exos. After 2 weeks, the neovascularization in Matrigel was measured and mean blood vessel density (MVD) was calculated. The above experiments were all controlled by the same amount of PBS. After identification, the cultured cells were consistent with the characteristics of ADSCs. ADSC-Exos were circular or elliptical membranous vesicle with uniform morphology under transmission electron microscopy, and expresses the signature proteins Alix and CD63 with particle size ranging from 30 to 200 nm. Confocal microscopy results showed that ADSC-Exos could be absorbed by HUVECs. The CCK-8 analysis showed that the cell proliferation of the experimental group was better than that of the control group at each time point ( P<0.05). The result of Transwell showed that the trans-membrane migration cells in the experimental group were significantly more than that in the control group ( t=9.534, P=0.000). In vitro, Matrigel tube-forming experiment showed that the number of tube-like structures in the experimental group was significantly higher than that of the control group ( t=15.910, P=0.000). In vivo, the MVD of the experimental group was significantly higher than that of the control group ( t=16.710, P=0.000). The ELISA assay showed that the expression of VEGF protein in the supernatant of the experimental group was significantly higher than that of the control group ( t=21.470, P=0.000). ADSC-Exos can promote proliferation, migration, and tube-like structure formation of HUVECs, suggesting that ADSC-Exos can promote angiogenesis in vitro and in vivo.

Metal oxide-confined interweaved titania nanotubes M/TNT (M = Mn, Cu, Ce, Fe, V, Cr, and Co) for the selective catalytic reduction of NOx in the presence of excess oxygen

A series of metal oxide confined titania nanotube M/TNT (M = Mn, Cu, Ce, Fe, V, Cr and Co) catalytic formulations were successfully synthesized using hydrothermal method followed by wet impregnation technique. The resulting materials were investigated for the low temperature Selective Catalytic Reduction (SCR) of NOx with NH3 in the presence of excess oxygen (10 vol.%). The prepared catalysts exhibit remarkable deNOx potential at temperatures as low as 100 °C and in a wide temperature range from 100 to 350 °C. Compared to manganese oxide supported on conventional TiO2, the synthesized Mn/TNT catalyst exhibits superior activity. Interestingly, the surface texture and tubular morphology of Mn/TNT catalyst greatly promotes the NOx conversion in the temperature regime of 100–300 °C. Our BET results of the as-prepared catalysts demonstrated that this is an effective synthesis for the generation of high specific surface area (421 m2/g) titania nanotubes. High resolution transmission electron microscopy (HR-TEM) results revealed the formation of tubular structure and multi-walled layer construction in the pristine titania nanotube (TNT), Mn/TNT and Ce/TNT samples. The relative atomic percentage value of Mn4+/Mn3+ and Ce3+/Ce4+ characterized by deconvoluted XPS spectra was considerably high for the Mn/TNT (Mn4+/Mn3+ = 2.15) and Ce/TNT catalysts (Ce3+/Ce4+ = 0.79), respectively. The existence of abundant surface Mn4+ species apparently contributes to the remarkable low-temperature SCR activity over our Mn/TNT catalyst. The H2-TPR results are in good accordance with our XPS results, with MnO2 being the prevailing phase and the increased reduction potentials of manganese appear to be the reason for the high deNOx activity of the Mn/TNT catalyst at low-temperatures. Vanadia based catalyst (V/TNT) formulations exhibit monolayer isolated species present over the support where V5+ is the dominant oxidation state. Titania nanotubes confined vanadium oxide catalyst demonstrates a broad operation temperature window attributed to the high dispersion of active species on the support.

17β-Estradiol Promotes Angiogenesis of Rat Cardiac Microvascular Endothelial Cells In Vitro.

BackgroundThe formation of new blood vessels, known as angiogenesis, is critical for recovery from ischemic heart disease, and estrogen is considered an important factor in this process. Here, we investigated the effects of 17β-estradiol (17β-E2) on proliferation and migration of cardiac microvascular endothelial cells (CMECs) in vitro.Material/MethodsRat CMECs were isolated and cultured with 17β-E2 (0.001–1 μmol/l) in the absence or presence of the estrogen antagonist tamoxifen. Then, the expression level of estrogen receptor alpha was evaluated by using immunofluorescence assay, RT-PCR, and Western blot. Cell proliferation was detected by methyl thiazolyl tetrazolium analysis and the cell migration was verified by a scraping assay and quantified by a Transwell chamber assay. CMEC differentiation was examined using a tube formation assay. Vascular endothelial growth factor (VEGF) secretion was detected by enzyme-linked immunosorbent assay.ResultsCMECs exhibited homogenous, polygonal, exhibited contact inhibition, and had characteristically ovoid nuclei with 1 or 2 nucleoli, and the cytoplasm exhibited red fluorescence after staining for von Willebrand factor. 17β-E2 treatment upregulated estrogen receptor alpha expression in CMECs. 17β-E2 treatment significantly promoted the proliferation, migration, tubular structure formation, and VEGF secretion in CMECs. The maximal proliferation occurred in the presence of 0.01 μmol/l 17β-E2. Furthermore, estrogen and VEGF were found to synergistically stimulate angiogenesis.ConclusionsOur data show that 17β-E2 promotes angiogenesis in vitro and suggests that estrogen treatment as a novel therapeutic modality in the management of arterial insufficiency.

The Growth of Carbon Nanotubes via Chemical Vapor Deposition Method; its Purification and Functionalization

Objectives: The main motive behind proposed work is to enhance the solubility of CNTs in aqueous medium and in various other solvents. Purification and functionalization helped in accomplishing aforementioned task. Methods/Statistical Analysis: Initially, CNTs were grown by Chemical Vapor Deposition method. Nickel metal catalyst solution was prepared and loaded onto SiO2 wafer, which is kept in vacuum assisted quartz tube. Acetylene was used as a hydrocarbon source and at 750 oC it decomposes at nickel metal surface resulting in growth of Carbon Nanotubes. Several characterizations such as SEM, EDX, XRD and FTIR were performed for confirmation of CVD grown CNTs. SEM confirms the formation of tubular structure i.e. carbon nanotube and EDX illustrates the presence of carbon and metal catalytic impurities. Findings: Such impurities restrict their use in physical, chemical, biological and environmental systems, thus purification is required. Among all purification methods, acid treatment suits best for removal of metallic impurities. An acid mixture of H2SO4 and HNO3 in ratio of 3:1 was utilized to remove the impurities present. These acid purified CNTs now contain carboxylic acid groups attached over defects, sidewalls and at ends. However another major shortcoming of CNTs is their insolubility in various liquids and polymers. Improvements: Therefore, to improve the solubility and to allow bonding with polymer matrix, amide functionalization was carried out. By the effect of thionyl chloride, the -OH bond was replaced by –Cl group, which was further replaced by -NH moiety. Characterizations were carried out at each step of CNT processing. Moreover, these functionalized CNTs encourages us to extend the present work to quantify the effect of as grown and f-CNT on the average static and dynamic mechanical properties of CNT/epoxy nanocomposites in future.

MMP-2 and MMP-13 affect vasculogenic mimicry formation in large cell lung cancer.

Matrix metalloproteinases (MMPs) have critical functions in tumour vasculogenic mimicry (VM). This study explored the mechanisms underlying MMP‐13 and MMP‐2 regulation of tumour VM formation in large cell lung cancer (LCLC). In our study, laminin5 (Ln‐5) fragments cleaved by MMP‐2 promoted tubular structure formation by the LCLC cell lines H460 and H661 in three‐dimensional (3D) cultures. Transient up‐regulation of MMP‐13 or treatment with recombinant MMP‐13 protein abrogated tubular structure formation of H460 cells in 3D culture. Treated cells with Ln‐5 fragments cleaved by MMP‐2 stimulated EGFR and F‐actin expression. Ln‐5 fragments cleaved by MMP‐13 decreased EGFR/F‐actin expression and disrupted VM formation. MMP‐13 expression was negatively correlated with VM, Ln‐5 and EGFR in LCLC tissues and xenograft. In vivo experiments revealed that VM was decreased when the number of endothelium‐dependent vessels (EDVs) increased during xenograft tumour growth, whereas MMP‐13 expression was progressively increased. In conclusion, MMP‐2 promoted and MMP‐13 disrupted VM formation in LCLC by cleaving Ln‐5 to influence EGFR signal activation. MMP‐13 may regulate VM and EDV formation.

EFFECT OF VOLTAGE ON TIO2 NANOTUBES FORMATION IN ETHYLENE GLYCOL SOLUTION

The crystalline phase of the TiO2 nanotubes without further heat treatment were studied. The TiO2 nanotube arrays were produced by anodization of Ti foil at three different voltage; 10, 40, and 60 V in a bath with electrolytes composed of ethylene glycol (EG), ammonium fluoride (NH4F), and hydrogen peroxide (H2O2). The H2O2 is a strong oxidizing agent which was used as oxygen provider to increase the oxidation rate for synthesizing highly ordered and smooth TiO2 nanotubes. Anodization at voltage greater than 10 V leads to the formation of tubular structure where higher anodization voltage (~ 60 V) yield to larger tube diameter (~ 180 nm). Crystallinity of the nanotubes is improved as the voltage was increased. The transformation of amorphous to anatase can be obtained for as anodized TiO2 without any heat treatment. The Raman spectra results show the anodization at 40 V and 60 V gives anatase peak in which confirms the crystalline phase. The stabilization of the crystalline phase is due to the oxygen vacancies and ionic mobilities during the anodization at high voltage.Â

Hydrogen sulfide facilities production of nitric oxide via the Akt/endothelial nitric oxide synthases signaling pathway to protect human umbilical vein endothelial cells from injury by angiotensin II.

Angiotensin II (Ang II) has been reported as key in inducing endothelial cell injury, and endothelial cells may produce nitric oxide (NO) to protect themselves. However, the underlying mechanism remains elusive. Human umbilical vein endothelial cells (HUVECs) were divided into five treatment groups as follows: Normal control, Ang II, Ang II + sodium hydrosulfide [NaHS; hydrogen sulfide (H2S) donor], Ang II + Akt inhibitors + NaHS, and Ang II + endothelial nitric oxide synthases (eNOS) inhibitors + NaHS. Subsequently, cell viability, apoptosis, migration, proliferation and adhesion ability were determined. In addition, tubular structure formation was observed, and the NO and phosphorylation levels of Akt and eNOS were evaluated. Compared with the normal control group, Ang II treatment reduced the viability of HUVECs and increased the level of cell apoptosis (P<0.05). Furthermore, Ang II treatment inhibited the phosphorylation level of eNOS and Akt, as well as the generation of NO (P<0.05). H2S reversed the above‑mentioned effects significantly and increased cell proliferation, adhesion ability and promoted tubular structure formation (P<0.05); however, H2S did not reverse the impact of eNOS and Akt phosphorylation levels after being processed with Akt and eNOS inhibitors, which indicates that H2S is capable of protecting HUVECs via the eNOS/Akt signaling pathway (P<0.05). Thus, H2S stimulates the production of NO and protects HUVECs via inducing the Akt/eNOS signaling pathway.

Pro-angiogenic Role of Insulin: From Physiology to Pathology.

The underlying molecular mechanisms involve in the regulation of the angiogenic process by insulin are not well understood. In this review article, we aim to describe the role of insulin and insulin receptor activation on the control of angiogenesis and how these mechanisms can be deregulated in human diseases. Functional expression of insulin receptors and their signaling pathways has been described on endothelial cells and pericytes, both of the main cells involved in vessel formation and maturation. Consequently, insulin has been shown to regulate endothelial cell migration, proliferation, and in vitro tubular structure formation through binding to its receptors and activation of intracellular phosphorylation cascades. Furthermore, insulin-mediated pro-angiogenic state is potentiated by generation of vascular growth factors, such as the vascular endothelial growth factor, produced by endothelial cells. Additionally, diseases such as insulin resistance, obesity, diabetes, and cancer may be associated with the deregulation of insulin-mediated angiogenesis. Despite this knowledge, the underlying molecular mechanisms need to be elucidated in order to provide new insights into the role of insulin on angiogenesis.

Femtosecond filamentation of optical vortex in a medium with anomalous group velocity dispersion

Within the framework of the slowly varying wave approximation method, the pictures of spatio-temporal dynamics of femtosecond pulse in a ring beam with phase singularity were obtained during propagation in CaF 2 crystal with anomalous group velocity dispersion. We show formation of tubular structure of the beam and explain reasons of its appearance. Fluence and linear plasma concentration dependencies on propagation distance are analyzed.

Cilostazol에 의한 뇌혈관내피세포의 세포이동 증진 효과연구

Cilostazol is known to be a selective inhibitor of phosphodiesterase III and is generally used to treat stroke. Our previous findings showed that cilostazol enhanced capillary density through angiogenesis after focal cerebral ischemia. Angiogenesis is an important physiological process for promoting revascularization to overcome tissue ischemia. It is a multistep process consisting of endothelial cell proliferation, migration, and tubular structure formation. Here, we examined the modulatory effect of cilostazol at each step of the angiogenic mechanism by using human brain microvascular endothelial cells (HBMECs). We found that cilostazol increased the migration of HBMECs in a dose-dependent manner. However, it did not enhance HBMEC proliferation and capillary-like tube formation. We used a cDNA microarray to analyze the mechanisms of cilostazol in cell migration. We picked five candidate genes that were potentially related to cell migration, and we confirmed the gene expression levels by real-time PCR. The genes phosphoserine aminotransferase 1 (PSAT1) and CCAAT/enhancer binding protein β (C/EBPβ) were up-regulated. The genes tissue factor pathway inhibitor 2 (TFPI2), retinoic acid receptor responder 1 (RARRES1), and RARRES3 were down-regulated. Our observations suggest that cilostazol can promote angiogenesis by promoting endothelial migration. Understanding the cilostazol-modulated regulatory mechanisms in brain endothelial cells may help stimulate blood vessel formation for the treatment of ischemic diseases.

Controlled Tubular Unit Formation from Collagen Film for Modular Tissue Engineering.

Bottom-up or modular tissue engineering is one of the emerging approaches to prepare biomimetic constructs in vitro, involving fabrication of small tissue units as building blocks before assembling them into functional tissue constructs. Herein, we reported a microscale tissue engineering approach to generate tubular tissue units through cellular contractile force induced self-folding of cell-laden collagen films in a controllable manner. Self-folding of cell-laden collagen films was driven by film contraction resulted from intrinsic contractile property of adherent mammalian cells seeded in collagen films. We explored in detail independent effects of collagen gel concentration, cell density, and intrinsic cellular contractility on self-folding and tubular structure formation of cell-laden collagen films. Through both experiments and theoretical modeling, we further demonstrated the effectiveness of integrating ridge array structures onto the backside of collagen films in introducing structural anisotropy and thus controlling self-folding directions of collage films. Our approach of using ridge array structures to introduce mechanical anisotropy and thus promote tubular tissue unit formation can be extended to other biomaterial systems and thus provide a simple yet effective way to prepare tubular tissue units for modular tissue engineering applications.

Three dimensional tubular structure self-assembled by vascular mesenchymal cells at stiffness interfaces of hydrogels.

In this study, we report a rational and robust methodology to construct three dimensional (3D) tubular-structures solely by self-assembly of vascular mesenchymal cells (VMCs). Using the cell-laden hyaluronic acid hydrogel surrounded by cell-free gel with a higher stiffness, VMCs spontaneously migrated across the interface and assembled into 3D tubes, which composes of numerous cells. Based on turing instability which describes the reaction-diffusion processes of inhibitors and activators, this result of 3D tubular structure formation agrees with theoretical predictions from simulations of the reaction-diffusion of morphogens and cells under the initial conditions of patterned cell-laden hydrogel. We showed that this combination of theoretical prediction and experiments is able to produce multi-cellular 3D tubes with desired dimensions and determinate orientation in hydrogel mimicking the 3D features of tubular tissue. This work provides a reliable methodology for creating tubular structures with controllable sizes inside the 3D hydrogel through multi-cellular self-organization.

LJNK, an indoline-2,3-dione-based aminopeptidase N inhibitor with promising antitumor potency.

In our previous study, we found that LJNK showed potent aminopeptidase N (APN)-inhibitory activity. In the current study, we further evaluated the antitumor effects of LJNK both in vitro and in vivo. Enzyme experiments showed that LJNK showed better inhibitory activity than bestatin against APN both from human carcinoma cells' surface and from porcine kidney microsomes. In addition, LJNK could suppress rat aortic ring microvessel growth and HUVEC tubular structure formation, which showed its stronger antiangiogenesis effects than bestatin. [(3-[4,5-Dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide)] assay and clonogenic assay showed that LJNK suppressed cancer cell growth both in the short and the long term. Mice bearing H22 transplantation tumor proved its antitumor effects in vivo. Annexin V-fluorescein isothiocyanate/propidium iodide assay showed that LJNK could induce 28.1% PLC/PRF/5 cell apoptosis and the apoptotic pathway was probably identified by western blot. The above-mentioned results suggested that LJNK inhibited cell proliferation and angiogenesis, and induced apoptosis by decreasing APN activity.

Glipizide suppresses prostate cancer progression in the TRAMP model by inhibiting angiogenesis.

Drug repurposing of non-cancer drugs represents an attractive approach to develop new cancer therapy. Using the TRAMP transgenic mouse model, glipizide, a widely used drug for type 2 diabetes mellitus, has been identified to suppress prostate cancer (PC) growth and metastasis. Angiogenesis is intimately associated with various human cancer developments. Intriguingly, glipizide significantly reduces microvessel density in PC tumor tissues, while not inhibiting prostate cancer cell proliferation from the MTT assay and flow cytometry investigation. Moreover, glipizide inhibits the tubular structure formation of human umbilical vein endothelial cells by regulating the HMGIY/Angiopoietin-1 signaling pathway. Taken together, these results demonstrate that glipizide has the potential to be repurposed as an effective therapeutic for the treatment of PC by targeting tumor-induced angiogenesis.

Angiogenesis by EVGATA−4 is mediated by transfer of let‐7 miRNA family

BackgroundIt is well known that extracellular vesicles (EVs) from stem cells are responsible for mediating diverse bio‐effects through delivering bioactive molecules. Here, we investigated whether the angiogenesis by EVs released from mesenchymal stem cells (MSC) overexpressing GATA4 (EVGATA−4) is mediated by miRs let‐7 family.Methods and ResultsMSCs were transduced GATA‐4 (MSCGATA−4) using the murine stem cell virus retroviral expression system. The conditioned medium was collected from MSCGATA−4 and its control counterpart MSCnull. EVs were isolated using exosome precipitation solution. The data of MicroRNA‐seq of EVs shows that 358 miRs were found in EVs with 46 miRs significantly increased in EVGATA−4 compared to those in EVs derived from MSCnull (EVnull). let‐7 family not only have had a high reads in EVs, but also significantly upregulated in EVGATA−4 compared to that in EVnull. The number of tubular structure formation was more in human umbilical vein endothelial cells (HUVECs) treated with EVGATA−4 (30μg/300μl) than in those treated with EVnull. The cumulative length of sprout per spheroid was also greater in EVGATA−4‐treated HUVECs than EVnull‐treated cells. To further study the effect of EVs on blood vessel formation, a Matrigel containing EVs (100μg/plug) was subcutaneously injected into the abdomen of mice. Two weeks later, the Matrigel plugs were harvested. The cell number and hemoglobin were significantly increased in the plugs containing EVGATA−4 compared to the plugs containing EVnull. The real‐time imaging capture system recorded that EVs pre‐labeled with PKH26 could be internalized by HUVECs. The expression of let‐7f in HUVECs treated with EVGATA−4 was significantly upregulated. Silence of let‐7f in EVGATA−4 by transfecting with an inhibitor of let‐7f reduced the angiogenetic effect of EVGATA−4. Thrombospondin 1 (THBS1) is one of the potential target genes of let‐7f and an inhibitor of angiogenesis. The expression of THBS1 was significantly down‐regulated in HUVEC treated with EVGATA−4. ConclusionThe increased pro‐angiogenic capacity of EVGATA−4 is associated with let‐7 transfer into HUVECs and down‐regulation of the expression of THBS1 in HUVECs.Support or Funding InformationThis work was supported by the National Institutes of Health grants HL105176 and HL114654 (M. Xu).

Nanografted Substrata and Triculture of Human Pericytes, Fibroblasts, and Endothelial Cells for Studying the Effects on Angiogenesis.

For the successful treatment of severe wounds, angiogenesis must be induced to provide a sufficient blood supply to the wound site. There have been many studies on various structural and biochemical factors regulating angiogenesis, such as surface topography, surface modifications, angiogenic factors, and various cell types. However, there is a paucity of research on the effects of micro- and nanoscale topography and the application of pericytes in angiogenesis. In this study, we utilized nanoscale topography combined with a triculture system consisting of human pericytes, fibroblasts, and endothelial cells. We investigated the effects of the nanografted substrata and a triculture system on proangiogenic therapies in vitro. Human dermal fibroblasts and human umbilical vein endothelial cells were seeded with human pericytes from the placenta directly onto nanografted substrata composed of polydimethylsiloxane. We demonstrated that key elements of angiogenesis, including segment and capillary-like tubular structure formation, as well as the secretion of extracellular matrix, were increased by interaction between the nanografted substrata and the coculture containing pericytes. Thus, nanografted surfaces and triculture systems containing human pericytes, fibroblasts, and endothelial cells promote angiogenesis.

Synthesis of Carbon Nanotubes Using Azerbaijan’s Oil

Distillate fractions of Azerbaijan’s oil-gasoline, jet fuel and diesel were tested as a raw material for producing carbon nanotubes. Aerosol CVD technique was employed at atmospheric and low pressures and ferrocene was used as a catalyst. It is shown that at atmospheric pressures carbon nanotubes were formed only from a gasoline fraction. Lower pressure in the reactor during the synthesis process leads to formation of carbon tubular structure from both petroleum fractions-gasoline and jet fuel. Other modifications of carbon were grown at atmospheric pressure in a case used fraction jet fuel and diesel fraction. MWCNTs with diameters of 35 - 65 nm have been grown at lower pressure in the reactor using gasoline fraction. The diameter of the MWNTs grown at atmospheric pressure in the reactor was in the range of 80 - 215 nm and the length reached 6 microns after the purification process.

MARCKSL1 exhibits anti-angiogenic effects through suppression of VEGFR-2-dependent Akt/PDK-1/mTOR phosphorylation.

Myristoylated alanine-rich C kinase substrate-like 1 (MARCKSL1) plays a pivotal role in the regulation of apoptosis and has been shown to maintain antitumor and metastasis-suppressive properties. In the present study, we examined the effects of MARCKSL1 as a novel anti-angiogenic agent on the inhibition of angiogenesis-mediated cell migration. MARCKSL1 also reduced vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cell (HUVEC) proliferation, as well as capillary-like tubular structure formation in vitro. MARCKSL1 disrupted phosphorylation of vascular endothelial growth factor receptor-2 (VEGFR-2) in ovarian tumorigenesis. In addition, MARCKSL1 showed potent anti-angiogenic activity and reduced the levels of VEGF and hypoxia-inducible factor 1α (HIF-1α) expression, an essential regulator of angiogenesis. Consistently, MARCKSL1 decreased VEGF‑induced phosphorylation of the PI3K/Akt signaling pathway components, including phosphoinositide-dependent protein kinase 1 (PDK-1), mammalian target of rapamycin (mTOR), tuberous sclerosis complex 2 (TSC-2), p70 ribosomal protein S6 kinase (p70S6K), and glycogen synthase kinase 3β (GSK-3β) protein. Collectively, our results provide evidence for the physiological/biological function of an endothelial cell system involved in angiogenesis through suppression of Akt/PDK-1/mTOR phosphorylation by interaction with VEGFR-2.

Impact of ER520, a candidate of selective estrogen receptor modulators, on in vitro cell growth, migration, invasion, angiogenesis and in vivo tumor xenograft of human breast cancer cells

ER520, a derivative of indenoisoquinoline, is a patented compound. This study was designed to screen its biological properties and to evaluate its antineoplastic and antiangiogenic effect. Western blot was employed to monitor the ERα and ERβ protein expression in human breast cancer MCF-7 cells and endometrial carcinoma Ishikawa cells. MTT assay was employed to determine cell proliferation. Cell adhesion, scratch and Transwell assay were utilized to estimate the ability of cellular adhesion, migration and invasion. ELISA kit was applied to detect the VEGF products in culture medium. In addition, the inhibitory effect of ER520 on the vessel-like construction of HUVEC cells and the angiogenesis of chicken embryos was investigated. The efficiency of ER520 on tumor growth in nude mice was also assessed. ER520 inhibited the expression of ERα in MCF-7 and Ishikawa cells, while it increased ERβ protein level. ER520 also suppressed the proliferation of MCF-7 and Ishikawa cells. Due to its remarkably negative role in cell adhesion, migration and invasion, ER520 showed a potential ability of inhibiting tumor metastasis. Meanwhile, ER520 reduced the VEGF secretion of MCF-7 and Ishikawa cells, prevented the formation of VEGF-stimulated tubular structure and the cell migration of HUVEC cells, and inhibited the angiogenesis of chicken chorioallantoic membrane. Animal experiment also demonstrated that ER520 could frustrate the in vivo tumor growth and the inhibitory ratio was 48.5 % compared with control group. Our findings indicate that ER520 possesses the competence to be a candidate against breast cancer and angiogenesis.

Reconstruction of lymphatic vessels in the mouse tail after cupping therapy.

The aim of the study was to investigate the regulatory mechanism of local lymphatic reconstruction after cupping therapy in a mouse model. The lymphatic reconstruction process in the mouse tail after cupping therapy as well as the expression levels of the vascular endothelial identification molecule CD34, prospero homeobox protein 1 (PROX1), and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1) were investigated for a duration of 4 days through immunohistochemistry experiments. On day 1 after cupping therapy, the CD34+ and LYVE-1+ cell densities were significantly increased, and the formed CD34+LYVE-1+ tubular structure started to express PROX1. This was followed by a decrease in both the CD34+ and LYVE-1+ stem cell densities to basal levels on the second day after cupping therapy. Both the CD34+ and LYVE-1+ cell densities subsequently increased again on the third day after cupping therapy. The increase in the LYVE-1+ density was accompanied by tubular structure formation, which is characteristic of lymphangiogenesis. In addition, the colocalisation of CD34+ and LYVE-1+ cells by immunohistochemistry suggests that the CD34+ stem cells differentiated into new lymphatic endothelial cells. Our findings indicate that the mechanism underlying the therapeutic effect of cupping therapy involves upregulation of vascular and lymphatic endothelial markers (CD34+, LYVE-1+, and CD34+LYVE-1+) in local tissues, which in turn promotes local new lymphatic vessel formation through the expression of PROX1.