12 research outputs found
Downregulation of cancer-associated fibroblast exosome-derived miR-29b-1-5p restrains vasculogenic mimicry and apoptosis while accelerating migration and invasion of gastric cancer cells via immunoglobulin domain-containing 1/zonula occluden-1 axis
Background: Cancer-associated fibroblast (CAF) exosomal miRNAs have gradually a hot spot in cancer therapy. This study mainly explores the effect of CAF-derived exosomal miR-29b-1-5p on gastric cancer (GC) cells. Methods: CAFs and exosomes were identified by Western blot and transmission electron microscopy. CAF-derived exosomes-GC cells co-culture systems were constructed. Effects of CAF-derived exosomal miR-29b-1-5p on GC cells were determined by cell counting kit-8, flow cytometry, wound healing, Transwell assays and Western blot. The relationship between miR-29b-1-5p and immunoglobulin domain-containing 1 (VSIG1) was assessed by TargetScan, dual-luciferase reporter and RNA immunoprecipitation (RIP) experiments. The interaction between VSIG1 and zonula occluden-1 (ZO-1) was detected by co-immunoprecipitation. Expressions of miR-29b-1-5p, VSIG1 and ZO-1 were determined by quantitative real-time PCR. Vascular mimicry (VM) was detected using immunohistochemistry and tube formation assays. Rescue experiments and xenograft tumor assays were used to further determine the effect of CAF-derived exosomal miR-29b-1-5p/VSIG1 on GC. Results: VM structure, upregulation of miR-29b-1-5p, and downregulation of VSIG1 and ZO-1 were shown in GC tissues. MiR-29b-1-5p targeted VSIG1, which interacted with ZO-1. CAF-derived exosomal miR-29b-1-5p inhibitor suppressed the viability, migration, invasion and VM formation, but promoted the apoptosis of GC cells. MiR-29b-1-5p inhibitor increased levels of VSIG1, ZO-1 and E-cadherin, whilst decreasing levels of VE-cadherin, N-cadherin and Vimentin in vitro and in vivo, which however was partially reversed by shVSIG1. Downregulation of CAF-derived exosomal miR-29b-1-5p impeded GC tumorigenesis and VM structure in vivo by upregulating VSIG1/ZO-1 expression. Conclusion: Downregulation of CAF-derived exosomal miR-29b-1-5p inhibits GC progression via VSIG1/ZO-1 axis.</p
Additional file 4: Figure S3. of Analysis of the ways and methods of signaling pathways in regulating cell cycle of NIH3T3 at transcriptional level
The interaction network between signaling pathway and cell cycle network. (DOC 9794 kb
Additional file 1: Table S1. of Analysis of the ways and methods of signaling pathways in regulating cell cycle of NIH3T3 at transcriptional level
Expression profiles of cell cycle-associated genes. (XLSX 163 kb
Additional file 2: Figure S1. of Analysis of the ways and methods of signaling pathways in regulating cell cycle of NIH3T3 at transcriptional level
Physiological activity that the reported genes involved. (DOC 580 kb
Additional file 3: Figure S2. of Analysis of the ways and methods of signaling pathways in regulating cell cycle of NIH3T3 at transcriptional level
Signal transduction of the signaling pathway that the reported genes involved (DOC 4343 kb
The novel protein C9orf116 promotes rat liver cell line BRL-3A proliferation - Fig 7
<p>Effect of C9orf116 on mitotic entry (A) EdU incorporation assays. The Click-it reaction revealed EdU staining (red). Cell nuclei were stained with Hoechst 33342 (blue). (B) The percentage of EdU-positive cells was quantified. (C) The expression of p-H3 (Ser10) was evaluated by Western blot analysis in C9orf116 knockdown and overexpression BRL-3A cells compared to their control, respectively. (D) The expression of p-H3 (Ser10) was quantified. The experiments were performed three times and three replicates in each one. The data were shown as the means ± SD. Representative images were shown *<i>P</i><0.05, **<i>P</i><0.01.</p
Effect of C9orf116 on apoptosis of BRL-3A cells.
<p>(A) Representative graphs obtained by flow cytometric analysis after double-staining with Annexin V-PE/7-AAD. (B) The percentage of late and early apoptotic cells were summed to give the total number of apoptotic cells. The experiments were performed three times and three replicates in each one. The data were shown as the means ± SD.</p
Effect C9orf116 on the expression of cell proliferation related genes.
<p>(A) The expression of cell proliferation related genes in C9-siR2 group at the mRNA level by qRT-PCR analysis. (B) The expression of cell proliferation related genes in C9-siR2 group at the protein level by western blot analysis. (C) The expression of cell proliferation related genes in pCDH-C9 group at the mRNA level by qRT-PCR analysis. (D) The expression of cell proliferation related genes in pCDH-C9 group at the protein level by western blot analysis. The experiments were performed three times and three replicates in each one, the data were shown as the means ± SD. Representative images were shown *<i>P</i><0.05, **<i>P</i><0.01.</p