Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hypoxic exosomes facilitate angiogenesis and metastasis in esophageal squamous cell carcinoma through altering the phenotype and transcriptome of endothelial cells

doi: 10.1186/s13046-019-1384-8

Figure Lengend Snippet: Uptake of exosomes derived from ECA109 and KYSE410 by HUVECs at 15 min, 60 min, 2 h and 4 h. HUVECs were cultured with exosomes (25 μg /mL) from ECA109, or exosomes (25 μg /mL) from KYSE410, or in the absence of exosomes (Exosome (−)). Fluorescence microscopy images showing the internalization of exosomes by HUVECs. Blue: Nucleus stained with DAPI. Red: PKH26-labeled exosomes. Green: Phalloidin-iFluor 488 Reagent. Scale bar, 50 μm

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) were also purchased from American Type Culture Collection and maintained in endothelial cell medium (ECM) (Science cell, USA).

Techniques: Derivative Assay, Cell Culture, Fluorescence, Microscopy, Staining, Labeling

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hypoxic exosomes facilitate angiogenesis and metastasis in esophageal squamous cell carcinoma through altering the phenotype and transcriptome of endothelial cells

doi: 10.1186/s13046-019-1384-8

Figure Lengend Snippet: The regulatory role of normoxic and hypoxic exosomes in the proliferation, cell cycle distribution, migration and invasion of HUVECs. HUVECs were cultured with exosomes (25 μg /mL) from ECA109 that cultured in normoxic environment (norm-Exo (ECA109)) or hypoxic environment (hypo-Exo (ECA109)), or exosomes (25 μg /mL) from KYSE410 that cultured in normoxic environment (norm-Exo (KYSE410)) or hypoxic environment (hypo-Exo (KYSE410)), or in the absence of exosomes (Exosome (−)). The proliferation of HUVECs was detected by colony formation assay ( a ). The graph summarizes the results of three independent experiments ( b ). The cell cycle of HUVECs were analyzed by flow cytometry. Representative pictures of the cell cycle distributions in HUVECs ( c ). The graph summarizes the results of three independent experiments ( d ). Transwell assays were used to investigate the migratory ( e ) and invasive ( g ) abilities of HUVECs. The graph summarizes the results of three independent experiments of migration ( f ) and invasion assay ( h ). Data was presented as mean ± standard deviation (SD).* P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) were also purchased from American Type Culture Collection and maintained in endothelial cell medium (ECM) (Science cell, USA).

Techniques: Migration, Cell Culture, Colony Assay, Flow Cytometry, Invasion Assay, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hypoxic exosomes facilitate angiogenesis and metastasis in esophageal squamous cell carcinoma through altering the phenotype and transcriptome of endothelial cells

doi: 10.1186/s13046-019-1384-8

Figure Lengend Snippet: Hypoxic exosomes promoted angiogenesis in vitro and increased the vessel density in vivo. HUVECs were plated with matrigel and cultured with exosomes (25 μg /mL) or not. Representative pictures of tube formation were taken after stained with Calcein-AM ( a ). The tube formation ability was quantified by measuring the total branching length ( b ). Matrigel containing exosomes, or not, were injected subcutaneously into the nude mice. Representative images of the general observation of matrigel plugs ( c ). In vivo neovascularization induced by exosomes was measured by H&E staining. Representative pictures of neovascularization were shown in ( d ) and quantified for blood vessel density ( e ). Data was presented as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) were also purchased from American Type Culture Collection and maintained in endothelial cell medium (ECM) (Science cell, USA).

Techniques: In Vitro, In Vivo, Cell Culture, Staining, Injection, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hypoxic exosomes facilitate angiogenesis and metastasis in esophageal squamous cell carcinoma through altering the phenotype and transcriptome of endothelial cells

doi: 10.1186/s13046-019-1384-8

Figure Lengend Snippet: Microarray analysis revealed differentially expressed RNAs between different groups. a Scatter-Plot of differentially expressed RNAs variations between HUVECs in the control group and norm-Exo group. Dots above the top line (red) and below the bottom line (green) indicated the fold change of the RNAs is more than 1.5 between the two groups. Heat map of the dysregulated mRNA, lncRNA and circular RNA expression in control group and norm-Exo group. b Scatter-Plot of differentially expressed RNAs variations between HUVECs in the control group and hypo-Exo group. Heat map of the dysregulated mRNA, lncRNA and circular RNA expression in control group and hypo-Exo group. Eight hundred and thirty nine down-regulated mRNAs ( c ), 113 up-regulated mRNAs ( d ), 232 down-regulated lncRNAs ( e ), 99 up-regulated lncRNAs ( f ), 692 down-regulated circular RNAs ( g ) and 86 up-regulated circular RNAs ( h ) were identified according to the intersection of transcriptome between norm-Exo group and hypo-Exo group

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) were also purchased from American Type Culture Collection and maintained in endothelial cell medium (ECM) (Science cell, USA).

Techniques: Microarray, Control, RNA Expression