title: Alginate-embedded HuH-7 cells increaseMMP-9and reduceOCLNexpression in vitro.
abstract: BACKGROUND
  Hepatocellular carcinoma is a common cancer, ranking third in cancer-associated deaths. An important cause of cancer patients' mortality is metastasis. At the start of metastasis progression, there is an epithelial-mesenchymal transition, characterized by matrix degradation, junction reductions and vessels formation. HuH-7 is a cell line used in research as an in vitro model for hepatocellular carcinoma. It is known that two-dimensional growth reflects tumor characteristics poorly. In contrast, three-dimensional cultures provide a better approach to the study of tumorigenic potential. The purpose of this work was to mimic a three-dimensional environment in order to assess gene expression of some epithelial-mesenchymal transition and metastasis progression markers in HuH-7 cells and compare them with traditional two-dimensional culture model.
  
  
  METHODS
  HuH-7 cells were encapsulated in sodium alginate (three-dimensional model) to be compared with cells grown in two-dimensional flasks. After 4 days in culture, gene expression ofMatrix metallopeptidase 9,Occludin, p65,Intercellular adhesion molecule 1andVascular endothelial growth factor Awas analyzed byqPCR and cytoskeleton assessment was performed by rhodamine-phalloidin staining.
  
  
  RESULTS
  Differences were found in gene expression, with a high increment ofMatrix metallopeptidase 9andOccludinreduction. The cytoskeleton morphology also showed differences, with a cytoplasm restricted only near the nuclei in the three-dimensional model.
  
  
  CONCLUSIONS
  This work shows the effects of using sodium alginate capsules as a three-dimensional model to the study of HuH-7. Cells in this 3D system show key markers of epithelial-mesenchymal transition, such asMatrix metallopeptidase 9overexpression andOccludindown-regulation.
authors.affiliation: Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil ; Post-Graduation Program in Child and Adolescent Health, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Virginia Andrea
authors.initials: VA
authors.lastname: Angiolini
authors.affiliation: Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil.
authors.forename: Carolina Uribe
authors.initials: CU
authors.lastname: Cruz
authors.affiliation: Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil ; Post-Graduation Program in Genetics and Molecular Biology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Mónica Luján
authors.initials: ML
authors.lastname: López
authors.affiliation: Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil.
authors.forename: Laura
authors.initials: L
authors.lastname: Simon
authors.affiliation: Gene Therapy Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS 90035-903 Brazil ; Post-Graduation Program in Child and Adolescent Health, Federal University of Rio Grande do Sul, Porto Alegre, Brazil ; Post-Graduation Program in Genetics and Molecular Biology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Ursula
authors.initials: U
authors.lastname: Matte
keywords: 3D culture; Encapsulation; Epithelial-mesenchymal transition; HuH-7