title: CD97 promotes gastric cancer cell proliferation and invasion through exosome-mediated MAPK signaling pathway.
abstract: OBJECTIVE
  To investigate the mechanism underlying the promoting role of CD97 in gastric cancer cell proliferation and invasion.
  
  
  METHODS
  Two types of exosomes released by gastric cancer cells with high (SGC/wt) or low (SGC/kd) CD97 expression were isolated by ultracentrifugation and identified by electron microscopy and western blot analysis. The influences of the two exosomes on gastric cancer cell proliferation and invasion were investigated by proliferation and Matrigel invasion assays. Exosomal miRNAs were subsequently isolated from the two samples and their miRNA profiles were compared via microarray assay analysis. Reverse transcription-quantitative real-time polymerase chain reaction was used to validate the microarray assay. Target genes of the differently expressed microRNAs were predicted based on five independent algorithms and were then subjected to gene oncology enrichment and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. After identifying the pathway that was the most likely altered, tumor cells were treated with the two exosomes at different concentrations, and the pathway activation was identified through western blot analysis.
  
  
  RESULTS
  Exosomes isolated from SGC/wt cells significantly promoted tumor cell proliferation in a dose-dependent manner in vitro. SGC/wt exosomes also significantly elevated the invasiveness of both SGC/wt (129.67 ± 8.327 vs 76.00 ± 5.292, P < 0.001) and SGC/kd (114.52 ± 9.814 vs 45.73 ± 4.835, P < 0.001) cells as compared to the exosomes released by SGC/kd cells. Microarray assay of the two exosomes revealed that 62 miRNAs were differently regulated with a signal intensity of > 500 and a false discovery rate < 0.05. The following KEGG analysis defined the MAPK signaling pathway as the most likely candidate pathway that regulated tumor cell proliferation and invasion. Through western blot analysis, significant up-regulations of phosphorylated MAPKs, including extracellular signal-regulated kinase, Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase, were detected in a dose-dependent manner in the SGC/wt exosomes treated groups, confirming activation of the MAPK signaling pathway stimulated by SGC/wt exosomes.
  
  
  CONCLUSIONS
  CD97 promotes gastric cancer cell proliferation and invasion in vitro through exosome-mediated MAPK signaling pathway, and exosomal miRNAs are probably involved in activation of the CD97-associated pathway.
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Li
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Da-Ren
authors.initials: DR
authors.lastname: Liu
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Guo-Gang
authors.initials: GG
authors.lastname: Li
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Hou-Hong
authors.initials: HH
authors.lastname: Wang
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Xiao-Wen
authors.initials: XW
authors.lastname: Li
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Yu-Lian
authors.initials: YL
authors.lastname: Wu
authors.affiliation: Chao Li, Da-Ren Liu, Guo-Gang Li, Hou-Hong Wang, Xiao-Wen Li, Yu-Lian Wu, Li Chen, Department of Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310021, Zhejiang Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D019295:Computational Biology; D004789:Enzyme Activation; D055354:Exosomes / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D006801:Humans; D020935:MAP Kinase Signaling System; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D060888:Real-Time Polymerase Chain Reaction; D043562:Receptors, G-Protein-Coupled; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D014162:Transfection
chemical_list: C095004:ADGRE5 protein, human; D015703:Antigens, CD; D035683:MicroRNAs; D043562:Receptors, G-Protein-Coupled; D020929:Mitogen-Activated Protein Kinase Kinases
keywords: CD97; Exosome; Gastric cancer; Invasion; Proliferation; miRNA