scholarly journals Nuclear Magnetic Resonance-Based Metabolomic Analysis of the Anticancer Effect of Metformin Treatment on Cholangiocarcinoma Cells

2020 ◽  
Vol 10 ◽  
Author(s):  
Jin Zhang ◽  
Caihua Hang ◽  
Ting Jiang ◽  
Shenghui Yi ◽  
Wei Shao ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Cancer Therapies ◽  
Metformin Treatment ◽  
Anticancer Effect ◽  
Multivariate Statistical ◽  
Cycle Arrest ◽  
Diabetes Drug ◽  
Umbilical Vein Endothelial Cells ◽  
And Control

Metformin is a widely prescribed anti-diabetes drug with potential utilities for cancer therapies. Several studies have related metformin to the reduced risk of cholangiocarcinoma (CCA), highlighting its potentialities for the treatments of CCA. However, the underlying molecular mechanisms remain elusive. Here, we demonstrated that metformin treatment could inhibit proliferations of the human CCA cell lines Mz-ChA-1 and QBC939 in dose-dependent manners. The NMR-based metabonomic analyses showed distinct discriminations between the metformin-treated (Met) and control (Ctrl) groups of both CCA cells. Characteristic metabolites were identified by a combination of multivariate statistical analysis of 1D 1H-NMR spectral data and the pair-wise t-test of metabolite levels. We then identified four significantly altered metabolic pathways based on the characteristic metabolites, including glucose metabolism, oxidative stress-related metabolism, energy metabolism, and amino acids metabolism. Comparing CCA cells with normal human umbilical vein endothelial cells (HUVECs), we found that metformin treatment profoundly promoted glycolysis and specifically increased the levels of BCAAs and UDP-GlcNAc, implying the occurrence of autophagy and cell cycle arrest in metformin-treated CAA cells. This work provides a mechanistic understanding of the anticancer effect of metformin treatment on CAA cells, and is beneficial to further developments of metformin as an anticancer drug.

scholarly journals NMR-Based metabolomic analysis of the anticancer effect of metformin on cholangiocarcinoma cells

2020 ◽  
Author(s):  
Jin Zhang ◽  
Caihua Hang ◽  
Wei Shao ◽  
Wengang Li ◽  
Donghai Lin
Keyword(s):  
Amino Acids ◽  
Tca Cycle ◽  
Essential Amino Acids ◽  
Cancer Therapies ◽  
Dependent Manner ◽  
Metformin Treatment ◽  
Anticancer Effect ◽  
Multivariate Statistical ◽  
Cell Viability Assay ◽  
Viability Assay

Abstract Background Metformin is a widely prescribed anti-diabetes drug with potential utilities for cancer therapies. Several previous studies have related metformin to the reduced risk of cholangiocarcinoma (CCA), highlighting its potentialities for the treatments of CCA. Methods In this study, cell viability assay and colony formation assay were used to test the inhibition effect of metformin on Mz-ChA-1 cells. The NMR-based metabonomic analysis was conduct to compare the differences between the metformin-treated (Met) and control (Ctrl) groups of the Mz-ChA-1 cells. Significant metabolites were identified from multivariate statistical analysis of 1D 1 H-NMR spectral data, and differential metabolites were identified from the pair-wise t-test of the metabolite levels. Significantly altered metabolic pathways were identified based on characteristic metabolites which were determined by a combination of the significant metabolites and differential metabolites. Results Here, we demonstrated that metformin treatment could inhibit the proliferation of the CCA cell line Mz-ChA-1 in a dose-dependent manner. The NMR-based metabonomic analysis showed a distinct discrimination between the Met and Ctrl groups of the Mz-ChA-1 cells. Moreover, The Met group exhibited promoted glycolysis and suppressed TCA cycle compared with the Ctrl group. While metformin treatment decreased non-essential amino acids, it also increased essential amino acids and UDP-GlcNAc, implying the occurrence of autophagy and cell cycle arrest in metformin-treated CAA cells . Conclusions This work provides a mechanistic understanding of the anticancer effect of metformin on CAA, and is beneficial to the further development of metformin as an anticancer drug

Newly synthesized benzimidazoles inhibits vascular endothelial growth factor and matrix metalloproteinase-2 and -9 levels in prostate cancer cells

2021 ◽  
Vol 21 ◽  
Author(s):  
Suleyman Ilhan ◽  
Gamze Dilekci ◽  
Adem Guner ◽  
Hakan Bektas
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Imidazole Ring ◽  
Matrix Metalloproteinase 2 ◽  
Benzimidazole Derivatives ◽  
Prostate Cancer Cells ◽  
Protein Levels ◽  
Umbilical Vein Endothelial Cells

Background: Investigating the effects of newly synthesized agents on various molecular mechanisms to understand their mechanism of action is an important step of pre-clinical screening. Benzimidazoles are composed of a unique fused benzene and imidazole ring and have attracted great attention due to their broad bioactivities, including antitumor. Objective: In the current study, we reported the synthesis of novel benzimidazole derivatives and investigated the possible cytotoxic and anti-angiogenic effects on human prostate cancer and umbilical vein endothelial cells (HUVECs). Methods: MTT assay was used to assess cell viability. A scratch assay was conducted to monitor the migration of cells. mRNA expression levels of VEGF, MMP-2, and MMP-9 were evaluated using qPCR. Changes in protein levels were evaluated by western blotting. Results: Compound G1, having a chlorine moiety, showed a potent cytotoxic activity on both prostate cancer cells and HUVECs, and inhibited cell migration via decreasing the mRNA and protein levels of key angiogenesis-related molecules such as VEGF, MMP-2, and MMP-9. Conclusion: These results suggest that newly synthesized G1 may be a novel anti-angiogenic agent for prostate cancer treatment.

Abstract 103: MiR-4261 Controls Endothelial Cells Angiogenesis

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Qiulian Zhou ◽  
Dongchao Lv ◽  
Qi Sun ◽  
Ping Chen ◽  
Yihua Bei ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Molecular Mechanisms ◽  
Heart Diseases ◽  
Umbilical Vein ◽  
Tissue Perfusion ◽  
Tube Formation ◽  
Artery Disease ◽  
Incorporation Assay ◽  
Umbilical Vein Endothelial Cells

Myocardial infarction (MI) is among major causes of morbidity and mortality associated with coronary artery disease. Angiogenesis improves tissue perfusion and cardiac repair after MI. Therefore, angiogenesis is considered to be a novel therapeutic way for ischemic heart diseases. MicroRNAs (miRNAs, miRs) have been reported to play important roles in regulating post-ischemic neovascularization. The current study aims at investigating the role of miR-4261 in angiogenesis. We found that miR-4261 mimics increased, while miR-4261 inhibitors decreased the proliferation of human umbilical vein endothelial cells (HUVEC) using EdU incorporation assay (17.25%±1.31% vs 30.91%±0.92% in nc-mimics vs mir-4261-mimics, 17.91%±1.36% vs 8.51%±0.82% in nc-inhibitor vs mir-4261-inhibitor, respectively) and CCK-8 assays (0.84±0.04 vs 1.38±0.04 in nc-mimics vs mir-4261-mimics, 0.80±0.02 vs 0.72±0.01 in nc-inhibitor vs mir-4261-inhibitor, respectively). The wound healing assay showed that miR-4261 mimic transfection resulted in a significant increase in the migration of HUVEC compared to that of the negative controls while miR-4261 inhibition had the opposite effects. Tube formation assays showed that HUVEC transfected with miR-4261 mimics increased the number of tubes formed (57.25±2.56 vs 81.5±2.53 in nc-mimics vs mir-4261-mimics, respectively), while miR-4261 inhibitor-transfected cells had the opposite effect (56.55±0.45 vs 41.38±0.52 in nc-inhibitor vs mir-4261-inhibitor, respectively). These results indicate that miR-4261 play an important role in regulating angiogenesis. However, it remains unknown which target gene mediated the effects of miR-4261. Thus, it will be of great interest to further investigate the molecular mechanisms of miR-4261 in the proliferation, migration, and tube formation of HUVEC in vitro. MiR-4261 could be a potential therapeutic target to enhance angiogenesis.

Abstract 181: BET Bromodomain Inhibition Suppresses Endothelial Inflammation and Atherosclerosis

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Jonathan Brown ◽  
Qiong Duan ◽  
Gabriel Griffin ◽  
Ronald Paranal ◽  
Steven Bair ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Rna Polymerase Ii ◽  
Umbilical Vein ◽  
Ldl Receptor ◽  
Flow Chamber ◽  
Cancer Therapies ◽  
Functional Studies ◽  
Endothelial Inflammation ◽  
Umbilical Vein Endothelial Cells

Introduction The BET bromodomain-containing family of proteins (BRD2, BRD3, BRD4) are epigenetic readers that coactivate transcription. Recent evidence indicates that BETs promote carcinogenesis and inflammation in sepsis, while BET bromodomain inhibitors are promising anti-cancer therapies. However, the role of chromatin remodeling in atherosclerosis in general and through BETs in particular remains unknown. Hypothesis We hypothesized that BET bromodomain-containing proteins coactivate proinflammatory responses in the vasculature with functional effects that promote atherogenesis. Methods and Results BET bromodomain inhibition, achieved with the highly selective, small-molecule inhibitor JQ1 significantly reduced early atherosclerosis (12 weeks) in cholesterol-fed, LDL receptor-null mice. In pursuing mechanisms for this effect, we identified BET protein expression in mouse and human endothelial cells (ECs) as well as endothelium from human atherosclerotic plaque. Treating human umbilical vein endothelial cells (HUVECs) with either JQ1 or siRNA to BRD2 or BRD4 potently suppresses TNFα-induced expression of adhesion molecules (SELE, VCAM1) and chemokines (CCL2, CXCL8). In chromatin immunoprecipation studies, TNFα stimulation of ECs recruited BETs to adhesion molecule and chemokine promoters coincident with RNA polymerase II and cyclin T1 localization, without altering NF-κB recruitment. In functional studies, JQ1 suppressed 1) monocyte adhesion to TNFα-activated HUVECs, 2) leukocyte rolling on cremaster post-capillary venules (intravital microscopy); 3) leukocyte transmigration (parallel-plate flow chamber); and 4) monocyte recruitment in thioglycolate-induced peritonitis in vivo . Conclusions BET bromodomain-containing proteins are novel determinants of pro-inflammatory transcription in the endothelium. Targeting chromatin by BET bromodomain inhibition may be a therapeutic strategy to limit atherosclerosis and other disorders involving endothelial inflammation.

scholarly journals Hypoxic human umbilical vein endothelial cells induce activation of adherent polymorphonuclear leukocytes

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3705-3716 ◽  
Author(s):  
T Arnould ◽  
C Michiels ◽  
J Remacle
Keyword(s):  
Endothelial Cells ◽  
Superoxide Dismutase ◽  
Superoxide Anion ◽  
Calcium Concentration ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Superoxide Anion Production ◽  
Elastase Inhibitor ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Abstract Several pieces of evidence are reported for the accumulation of activated neutrophils in ischemic and reperfused tissues leading to the transformation of the ischemic tissue into an inflammatory territory and to an enhancement of tissue damages during reoxygenation. However, the molecular mechanisms responsible for these observations and the precise role played by endothelial cells in this process are still poorly understood. In this study, an in vitro model that mimics this situation was used to investigate the effects of hypoxia-incubated human umbilical vein endothelial cells (HUVEC) on polymorphonuclear leukocyte (PMN) functions. A strong PMN activation characterized by an increase in intracellular calcium concentration as well as by superoxide anion release and leukotriene B4 production was observed when these cells were coincubated with hypoxic HUVEC. On the other hand, conditioned medium from hypoxia-incubated HUVEC failed to activate PMN, as determined by the lack of PMN calcium concentration increase, the failure of superoxide anion production enhancement, as well as the absence of effects on the integrin CD18, CD11a, and CD11b expression. These results indicate that the presence of hypoxia- incubated HUVEC is necessary to obtain an activation of the PMN, probably via the adherence process. Once activated by coincubation with hypoxic HUVEC, PMN became cytotoxic, as evidenced by 51Cr released from prelabeled HUVEC. This cytotoxic effect of activated PMN for hypoxic endothelial cells could be prevented by a combination of superoxide dismutase and catalase (94% inhibition), whereas superoxide dismutase alone was inefficient. Antiprotease (alpha 2-macroglobulin) and a specific elastase inhibitor (MAAPV-CMK) were also inefficient. These results correlate very well with the fact that no increase in elastase release could be observed in supernatants from PMN coincubated with hypoxic HUVEC. Furthermore, when adherence process was blocked by oleic acid or by anti-ICAM-1 monoclonal antibodies, protection was, respectively, 90% and 72%. We thus evidenced that free radicals but not elastase released from activated PMN coincubated with hypoxic HUVEC are involved in HUVEC injury. We conclude from these results that PMN activation is initiated by PMN adherence to hypoxic HUVEC. These observations indicate that hypoxic HUVEC may be partly responsible for neutrophil activation observed in ischemic tissues, which is part of the amplification process of tissue damage.

scholarly journals Microcystins Induces Vascular Inflammation in Human Umbilical Vein Endothelial Cells via Activation of NF-κB

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Jun Shi ◽  
Jie Zhou ◽  
Min Zhang
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecules ◽  
Inflammatory Process ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Health Hazard ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Microcystins (MCs) produced by toxic cyanobacteria cause serious water pollution and public health hazard to humans and animals. However, direct molecular mechanisms of MC-LR in vascular endothelial cells (ECs) have not been understood yet. In this study, we investigated whether MC-LR induces vascular inflammatory process in cultured human umbilical vein endothelial cells (HUVECs). Our data demonstrated that MC-LR decreased HUVECs proliferation and tube formation and enhanced apoptosis. MC-LR also induced intracellular reactive oxygen species formation (ROS) in HUVECs. The MC-LR directly stimulated phosphorylation of NF-κB. Furthermore, MC-LR also increased cell adhesion molecules (ICAM-1 and VCAM-1) expression in HUVECs. Taken together, the present data suggested that MC-LR induced vascular inflammatory process, which may be closely related to the oxidative stress, NF-κB activation, and cell adhesion molecules expression in HUVECs. Our findings may highlight that MC-LR causes potential damage to blood vessels.

scholarly journals ZFAT is essential for endothelial cell assembly and the branch point formation of capillary-like structures in an angiogenesis model

2010 ◽  
Vol 15 (4) ◽  
Author(s):  
Yasuhiro Yoshida ◽  
Toshiyuki Tsunoda ◽  
Yasuo Takashima ◽  
Takahiro Fujimoto ◽  
Keiko Doi ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Branch Point ◽  
Molecular Mechanisms ◽  
Network Formation ◽  
Umbilical Vein ◽  
Susceptibility Gene ◽  
Cell Assembly ◽  
Autoimmune Thyroid ◽  
Umbilical Vein Endothelial Cells ◽  
Angiogenesis Model

AbstractZFAT, originally identified as a susceptibility gene for autoimmune thyroid disease, encodes a transcriptional regulator with one AT-hook and 18 C2H2-type zinc-finger domains. It is highly conserved among species. Here, we demonstrate that ZFAT is clearly expressed in human umbilical vein endothelial cells (HUVECs). Furthermore, we show that endothelial cell assembly and the branch point formation of capillary-like structures in HUVECs is impaired by the reduction of ZFAT expression through the use of ZFAT-miRNAs, whereas differences in cell proliferation or apoptotic features were not observed after the reduction in ZFAT expression. These results suggest that ZFAT may have critical roles in the capillary-like network formation that is involved in vascular remodeling. Elucidating the ZFAT-mediated transcriptional network will lead to a better understanding of the molecular mechanisms of angiogenesis.

scholarly journals Different RNA splicing mechanisms contribute to diverse infective outcome of classical swine fever viruses of differing virulence: insights from the deep sequencing data in swine umbilical vein endothelial cells

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2113 ◽  
Author(s):  
Pengbo Ning ◽  
Yulu Zhou ◽  
Wulong Liang ◽  
Yanming Zhang
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Rna Splicing ◽  
Molecular Mechanisms ◽  
Computational Study ◽  
Umbilical Vein ◽  
Classical Swine Fever ◽  
Pathway Enrichment Analysis ◽  
Splicing Regulation ◽  
Umbilical Vein Endothelial Cells

Molecular mechanisms underlying RNA splicing regulation in response to viral infection are poorly understood. Classical swine fever (CSF), one of the most economically important and highly contagious swine diseases worldwide, is caused by classical swine fever virus (CSFV). Here, we used high-throughput sequencing to obtain the digital gene expression (DGE) profile in swine umbilical vein endothelial cells (SUVEC) to identify different response genes for CSFV by using both Shimen and C strains. The numbers of clean tags obtained from the libraries of the control and both CSFV-infected libraries were 3,473,370, 3,498,355, and 3,327,493 respectively. In the comparison among the control, CSFV-C, and CSFV-Shimen groups, 644, 158, and 677 differentially expressed genes (DEGs) were confirmed in the three groups. Pathway enrichment analysis showed that many of these DEGs were enriched in spliceosome, ribosome, proteasome, ubiquitin-mediated proteolysis, cell cycle, focal adhesion, Wnt signalling pathway, etc., where the processes differ between CSFV strains of differing virulence. To further elucidate important mechanisms related to the differential infection by the CSFV Shimen and C strains, we identified four possible profiles to assess the significantly expressed genes only by CSFV Shimen or CSFV C strain. GO analysis showed that infection with CSFV Shimen and C strains disturbed ‘RNA splicing’ of SUVEC, resulting in differential ‘gene expression’ in SUVEC. Mammalian target of rapamycin (mTOR) was identified as a significant response regulator contributed to impact on SUVEC function for CSFV Shimen. This computational study suggests that CSFV of differing virulence could induce alterations in RNA splicing regulation in the host cell to change cell metabolism, resulting in acute haemorrhage and pathological damage or infectious tolerance.

scholarly journals Peptides Isolated from Yak Milk Residue Exert Antioxidant Effects through Nrf2 Signal Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-17
Author(s):  
Feiyan Yang ◽  
Xudong He ◽  
Tao Chen ◽  
Jinliang Liu ◽  
Zhang Luo ◽  
...  
Keyword(s):  
Protective Effect ◽  
Molecular Mechanisms ◽  
Umbilical Vein ◽  
Damage Model ◽  
Nrf2 Pathway ◽  
Yak Milk ◽  
Nrf2 Signaling Pathway ◽  
Protein Expressions ◽  
Induced Apoptosis ◽  
Umbilical Vein Endothelial Cells

Food-derived bioactive peptides are considered as the important sources of natural bioactive ingredients. Approximately 3094 peptides were identified by nESI-LC–MS/MS in the hydrolyzed yak milk residue. Peptide KALNEINQF (T10) is the strongest antioxidant peptide. The damage model of H2O2-induced human umbilical vein endothelial cells (HUVECs) was used to evaluate the antioxidant effect. After treatment with 25, 50, or 100 μg/mL T10 peptide, T10 obviously decreased H2O2-induced damage and increased the cell survival. Comparing with the H2O2-induced damage group, superoxide dismutase (SOD) activities were significantly increased 1.03, 1.1, and 1.33 times, and glutathione reductase (GR) activities were significantly increased 1.11, 1.30, and 1.43 times, respectively. Malondialdehyde (MDA) also reduced 1.41, 1.54, and 1.72 times, respectively. T10 inhibited H2O2-induced apoptosis in HUVECs, and protein expressions of the apoptosis-related genes bcl-2 and bax were increased and decreased by 1.95 and 1.44 times, respectively, suggesting T10 decreases apoptosis of the mitochondria-dependent pathway. Comparing with the H2O2-induced damage group, the RNA expressions of Nrf2, HO-1, and NQO1 were significantly increased by 2.00, 2.11, and 1.94 times; the protein expressions of p-Nrf2, HO-1, and NQO1 were significantly increased by 2.67, 1.73, and 1.04 times; and Keap1 was downregulated by 3.9 and 1.32 times, respectively. T10 also regulated the Nrf2 pathway and expressions of related genes (Keap1, HO-1, and NQO1), and blocking the Nrf2 pathway in the model decreased the protective effect of T10. Taken together, T10 peptide isolated from yak milk residue has a protective effect against H2O2-induced damage in HUVECs and the molecular mechanisms are involved in the regulation of Nrf2 signaling pathway and cell apoptosis.

scholarly journals Effect of flow on polymorphonuclear leukocyte/endothelial cell adhesion

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1284-1290 ◽  
Author(s):  
MB Lawrence ◽  
LV McIntire ◽  
SG Eskin
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Shear Stresses ◽  
Adhesive Interaction ◽  
Experimental Conditions ◽  
Coated Surface ◽  
Wall Shear ◽  
Umbilical Vein Endothelial Cells ◽  
And Control

Abstract The effect of flow on the adhesion of polymorphonuclear leukocytes (PMNL) to vascular endothelium was investigated using a parallel plate chamber with a well-defined flow field. Washed PMNL were perfused over a monolayer of primary human umbilical vein endothelial cells (HUVEC) pretreated with formyl-methionyl-leucyl-phenylalanine (FMLP, 1 X 10(-7) mol/L) for five minutes. In other experiments HUVEC were pretreated with interleukin 1 (IL1,2 U/mL) for four hours. PMNL adhesion to stimulated and control HUVEC was measured over a physiologic range of wall shear stresses. PMNL adhesion to nylon-coated surface was also studied. At a wall shear stress of 0.98 dynes/cm2,283 +/- 37.3 PMNL/mm2 (mean +/- SEM) adhered to FMLP-treated HUVEC while 195 +/- 20.3 PMNL/mm2 adhered to control HUVEC. At 1.96 dynes/cm2, 68 +/- 14.1 PMNL/mm2 adhered to FMLP-treated HUVEC and 42 +/- 6.0 PMNL/mm2 adhered to control HUVEC. At 3.92 dynes/cm2, virtually no PMNL adherence was noted on either control or FMLP-treated HUVEC. On IL 1-treated HUVEC at 1.96 dynes/cm2, 371 +/- 25.8 PMNL/mm2 adhered while 28 +/- 2.9 PMNL/mm2 adhered to control HUVEC. PMNL adhesion to IL 1-treated and control HUVEC dropped to 10.2 +/- 3.8 and 6.8 +/- 3.5 PMNL/mm2, respectively, at 3.01 dynes/cm2. The effect of flow on PMNL adhesion appears to be an important factor in determining the outcome of the PMNL/HUVEC adhesive interaction under these experimental conditions.

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