Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells

GATA4, a transcription factor crucial for early liver development, has been implicated in the pathophysiology of hepatoblastoma, an embryonal tumor of childhood. However, the molecular and phenotypic consequences of GATA4 expression in hepatoblastoma are not fully understood. We surveyed GATA4 expre...

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Main Authors: Tea Soini, Katja Eloranta, Marjut Pihlajoki, Antti Kyrönlahti, Oyediran Akinrinade, Noora Andersson, Jouko Lohi, Mikko P Pakarinen, David B Wilson, Markku Heikinheimo
Format: Article
Language:English
Published: SAGE Publishing 2018-08-01
Series:Tumor Biology
Online Access:https://doi.org/10.1177/1010428318785498
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author Tea Soini
Katja Eloranta
Marjut Pihlajoki
Antti Kyrönlahti
Oyediran Akinrinade
Noora Andersson
Jouko Lohi
Mikko P Pakarinen
David B Wilson
Markku Heikinheimo
author_facet Tea Soini
Katja Eloranta
Marjut Pihlajoki
Antti Kyrönlahti
Oyediran Akinrinade
Noora Andersson
Jouko Lohi
Mikko P Pakarinen
David B Wilson
Markku Heikinheimo
author_sort Tea Soini
collection DOAJ
description GATA4, a transcription factor crucial for early liver development, has been implicated in the pathophysiology of hepatoblastoma, an embryonal tumor of childhood. However, the molecular and phenotypic consequences of GATA4 expression in hepatoblastoma are not fully understood. We surveyed GATA4 expression in 24 hepatoblastomas using RNA in situ hybridization and immunohistochemistry. RNA interference was used to inhibit GATA4 in human HUH6 hepatoblastoma cells, and changes in cell migration were measured with wound healing and transwell assays. RNA microarray hybridization was performed on control and GATA4 knockdown HUH6 cells, and differentially expressed genes were validated by quantitative polymerase chain reaction or immunostaining. Plasmid transfection was used to overexpress GATA4 in primary human hepatocytes and ensuring changes in gene expression were measured by quantitative polymerase chain reaction. We found that GATA4 expression was high in most hepatoblastomas but weak or negligible in normal hepatocytes. GATA4 gene silencing impaired HUH6 cell migration. We identified 106 differentially expressed genes (72 downregulated, 34 upregulated) in knockdown versus control HUH6 cells. GATA4 silencing altered the expression of genes associated with cytoskeleton organization, cell-to-cell adhesion, and extracellular matrix dynamics (e.g. ADD3, AHNAK, DOCK8, RHOU, MSF, IGFBP1, COL4A2 ). These changes in gene expression reflected a more epithelial (less malignant) phenotype. Consistent with this notion, there was reduced F-actin stress fiber formation in knockdown HUH6 cells. Forced expression of GATA4 in primary human hepatocytes triggered opposite changes in the expression of genes identified by GATA4 silencing in HUH6 cells. In conclusion, GATA4 is highly expressed in most hepatoblastomas and correlates with a mesenchymal, migratory phenotype of hepatoblastoma cells.
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spelling doaj-art-b31ebeb5e8eb4d1f866deeb7fe85dc472025-08-20T03:35:58ZengSAGE PublishingTumor Biology1423-03802018-08-014010.1177/1010428318785498Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cellsTea Soini0Katja Eloranta1Marjut Pihlajoki2Antti Kyrönlahti3Oyediran Akinrinade4Noora Andersson5Jouko Lohi6Mikko P Pakarinen7David B Wilson8Markku Heikinheimo9Pediatric Research Center, Children’s Hospital, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandPediatric Research Center, Children’s Hospital, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandDepartment of Pediatrics, St. Louis Children’s Hospital, Washington University School of Medicine in St. Louis, St. Louis, MO, USADepartment of Pediatrics, St. Louis Children’s Hospital, Washington University School of Medicine in St. Louis, St. Louis, MO, USAPediatric Research Center, Children’s Hospital, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandPediatric Research Center, Children’s Hospital, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandDepartment of Pathology, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandUnit of Pediatric Surgery and Pediatric Liver and Gut Research Group, Children’s Hospital, University of Helsinki and Helsinki University Hospital, Helsinki, FinlandDepartment of Developmental Biology, Washington University School of Medicine in St. Louis, St. Louis, MO, USADepartment of Pediatrics, St. Louis Children’s Hospital, Washington University School of Medicine in St. Louis, St. Louis, MO, USAGATA4, a transcription factor crucial for early liver development, has been implicated in the pathophysiology of hepatoblastoma, an embryonal tumor of childhood. However, the molecular and phenotypic consequences of GATA4 expression in hepatoblastoma are not fully understood. We surveyed GATA4 expression in 24 hepatoblastomas using RNA in situ hybridization and immunohistochemistry. RNA interference was used to inhibit GATA4 in human HUH6 hepatoblastoma cells, and changes in cell migration were measured with wound healing and transwell assays. RNA microarray hybridization was performed on control and GATA4 knockdown HUH6 cells, and differentially expressed genes were validated by quantitative polymerase chain reaction or immunostaining. Plasmid transfection was used to overexpress GATA4 in primary human hepatocytes and ensuring changes in gene expression were measured by quantitative polymerase chain reaction. We found that GATA4 expression was high in most hepatoblastomas but weak or negligible in normal hepatocytes. GATA4 gene silencing impaired HUH6 cell migration. We identified 106 differentially expressed genes (72 downregulated, 34 upregulated) in knockdown versus control HUH6 cells. GATA4 silencing altered the expression of genes associated with cytoskeleton organization, cell-to-cell adhesion, and extracellular matrix dynamics (e.g. ADD3, AHNAK, DOCK8, RHOU, MSF, IGFBP1, COL4A2 ). These changes in gene expression reflected a more epithelial (less malignant) phenotype. Consistent with this notion, there was reduced F-actin stress fiber formation in knockdown HUH6 cells. Forced expression of GATA4 in primary human hepatocytes triggered opposite changes in the expression of genes identified by GATA4 silencing in HUH6 cells. In conclusion, GATA4 is highly expressed in most hepatoblastomas and correlates with a mesenchymal, migratory phenotype of hepatoblastoma cells.https://doi.org/10.1177/1010428318785498
spellingShingle Tea Soini
Katja Eloranta
Marjut Pihlajoki
Antti Kyrönlahti
Oyediran Akinrinade
Noora Andersson
Jouko Lohi
Mikko P Pakarinen
David B Wilson
Markku Heikinheimo
Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
Tumor Biology
title Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
title_full Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
title_fullStr Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
title_full_unstemmed Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
title_short Transcription factor GATA4 associates with mesenchymal-like gene expression in human hepatoblastoma cells
title_sort transcription factor gata4 associates with mesenchymal like gene expression in human hepatoblastoma cells
url https://doi.org/10.1177/1010428318785498
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