Journal: Stem Cell Research & Therapy

Article Title: microRNA-375 released from extracellular vesicles of bone marrow mesenchymal stem cells exerts anti-oncogenic effects against cervical cancer

doi: 10.1186/s13287-020-01908-z

Figure Lengend Snippet: Differential expressions of miR-375 and MELK were detected in cervical cancer. a The heat map of the top 50 differentially expressed genes in GSE7803 microarray. b The heat map of the top 50 differentially expressed genes in GSE63514 microarray. In panels a and b , the X -axis indicates the sample number while the Y -axis represents the gene. The tree diagram on the left indicates the gene expression cluster. Each square represents the expression of one gene in one sample. The histogram on the right shows intensity as a color gradation. c Intersection of differentially expressed genes in cervical cancer. Two circles represent the upregulated genes in cervical cancer-related two microarrays. The intersected region represents the intersection results. d Protein-protein intersection network of differentially expressed genes in cervical cancer. The circle reflects the core degree. e The expression of MELK in a sample at different stages of cervical cancer. The X -axis indicates the sample number while the Y -axis represents the gene. The first box indicates the MELK expression in normal cervical samples while the remaining four boxes present the MELK expression in cervical cancer samples at different stages. f Intersection of regulatory BMSC-EV-derived miRNAs and miRNAs in cervical cancer samples. The three circles represent the results obtained from the mirDIP database, TargetScan database, and previous literature, respectively. g The expression of miR-375 was determined using RT-qPCR in HcerEpic, CaSki, C33A, HeLa, and SiHa cell lines, normalized to U6. h The mRNA expression of MELK was determined using RT-qPCR in HcerEpic, CaSki, C33A, HeLa, and SiHa cell lines, normalized to β-actin. The measurement data are presented as mean ± SD. Multiple groups of data are compared by one-way ANOVA and Tukey’s test. * p < 0.05 compared with the HcerEpic cell line

Article Snippet: Human normal cervical epithelial cells (HcerEpic), human cervical cancer cell lines (CaSki, C33A, HeLa and SiHa), and HEK293T cells were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Microarray, Gene Expression, Expressing, Derivative Assay, Quantitative RT-PCR

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Representative immunofluorescent staining of cultured mesothelioma cells using AX10 antibody (a), immunohistochemical staining (b), and secondary antibody‐drug conjugate assay in vitro (c). (a) AX10 immunoreactivity in MPM‐1, −2, and −3 cells, representing sarcomatoid, epithelioid, and biphasic type mesothelioma, respectively. All MPM‐1, −2, and −3 cells exhibited AX10 antibody immunoreactivity at the cell surface. The staining was analyzed using a Guava easyCyte cell analyzer and accompanying software to obtain a one‐parameter log histogram. (b) AX10 immunoreactivity in various mesothelioma tissue specimens. Weak or no AX10 immunoreactivity was detected in five out of 10 epithelioid mesothelioma tissues (a). One out of five biphasic mesotheliomas exhibited AX10 immunoreactivity in spindle sarcomatoid components (arrow) but weak immunoreactivity in epithelioid components (arrowhead) (b). Five out of six sarcomatoid mesothelioma tissues exhibited strong AX10 immunoreactivity (c). Little AX10 immunoreactivity was detected in normal human tissues. No significant AX10 immunoreactivity was detected in the lung (d) (pleural mesothelial cells; insert) tissue specimens. Weak AX10 immunoreactivity was detected in myofibrous cells in the uterus (e). We did not detect any significant AX10 immunoreactivity in the brain, liver, or kidney, whereas strong AX10 immunoreactivity was observed in a nonmelanocytic (hypomelanocytic) melanoma tissue sample that was supplementally included in the microarray (f) (staining without AX10 antibody; insert). (c) MPM‐1 sarcomatoid mesothelioma cells were incubated with AX10 at 10, 100, and 1000 ng/mL followed by incubation with anti‐murine IgG (Fc) antibody conjugated to duocarmycin. Representative staining with Annexin V‐PI is presented. Note the dose‐dependent Annexin V‐positive and PI‐negative apoptotic MPM‐1 cells in the presence of AX10 antibody

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Staining, Cell Culture, Immunohistochemical staining, In Vitro, Software, Microarray, Incubation

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: AX10 does not affect cell proliferation, but significantly decreases Matrigel invasion activity of MPM‐1 sarcomatoid mesothelioma cells in vitro. (a) Representative cell proliferation assay. At 24 h, the cell number was 1.80 ± 0.10 (mock) and 1.77 ± 0.06 (AX10). Respective numbers at 48 h were 2.40 ± 0.10 (mock) and 2.37 ± 0.12 (AX10), while at 72 h they were 3.90 ± 0.20 (mock) and 4.20 ± 0.61 (AX10). The data represent means ± SD from triplicate assays (Student's t ‐test, p > 0.5). (b) AX10 significantly reduced Matrigel invasion activity of MPM‐1 cells (Student's t ‐test, p < 0.01). The number of invading cells was 59.7 ± 7.02 (mock) and 10.3 ± 1.52 (AX10) at 24 h, and 210.7 ± 11.4 (mock) and 15.0 ± 3.00 (AX10) at 48 h. Data from triplicate assays are expressed as means ± SD ( n = 3). (c) Cells that migrated to the lower surface of the membrane are shown (48 h). Original magnification, ×100

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Activity Assay, In Vitro, Proliferation Assay, Membrane

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Inhibitory effect of AX10 on MPM‐1 xenotransplanted sarcomatoid mesothelioma cell proliferation. (a) Inoculation of AX10 antibody delayed the growth of xenotransplanted MPM‐1 sarcomatoid mesothelioma tumors. On day 0, SCID‐NOD mice were subcutaneously implanted with MPM‐1 cells. The following day, day 3, the mice were administered AX10 antibody or vehicle only by intraperitoneal injection and weekly thereafter as indicated by arrows. Values are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test (* p < 0.01). (b) On day 42, the xenotransplanted tumors were excised to determine their weight. Total tumor weights are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test ( p < 0.01). (c) Gross and histological appearance of a representative xenotransplanted tumor. Arrowhead indicates the tumor without AX10 antibody, while the arrow indicates the small tumor remaining following weekly AX10 injection. Note the elimination of tumor cells, which were histologically replaced by regenerative muscle in mice inoculated with AX10 antibody. Scale bar indicates 100 μm

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Injection

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