Senescent human hepatocytes express a unique secretory phenotype and promote macrophage migration

Irvine, KM; Skoien, R; Bokil, NJ; Melino, M; Thomas, GP; Loo, D; Gabrielli, B; Hill, MM; Sweet, MJ; Clouston, AD; Powell, EE

Senescent human hepatocytes express a unique secretory phenotype and promote macrophage migration

Irvine, KM Skoien, R Bokil, NJ

Melino, M Thomas, GP Loo, D Gabrielli, B Hill, MM Sweet, MJ Clouston, AD Powell, EE

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Publication Type:: Journal Article
Citation:: World Journal of Gastroenterology, 2014, 20 (47), pp. 17851 - 17862
Issue Date:: 2014-01-01

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Field	Value	Language
dc.contributor.author	Irvine, KM	en_US
dc.contributor.author	Skoien, R	en_US
dc.contributor.author	Bokil, NJ https://orcid.org/0000-0003-3202-9249	en_US
dc.contributor.author	Melino, M	en_US
dc.contributor.author	Thomas, GP	en_US
dc.contributor.author	Loo, D	en_US
dc.contributor.author	Gabrielli, B	en_US
dc.contributor.author	Hill, MM	en_US
dc.contributor.author	Sweet, MJ	en_US
dc.contributor.author	Clouston, AD	en_US
dc.contributor.author	Powell, EE	en_US
dc.date.available	2014-07-16	en_US
dc.date.issued	2014-01-01	en_US
dc.identifier.citation	World Journal of Gastroenterology, 2014, 20 (47), pp. 17851 - 17862	en_US
dc.identifier.issn	1007-9327	en_US
dc.identifier.uri	http://hdl.handle.net/10453/118241
dc.description.abstract	© 2014 Baishideng Publishing Group Inc. All rights reserved. AIM: To develop a model of stress-induced senescence to study the hepatocyte senescence associated secretory phenotype (SASP). METHODS: Hydrogen peroxide treatment was used to induce senescence in the human HepG2 hepatocyte cell line. Senescence was confirmed by cytochemical staining for a panel of markers including Ki67, p21, heterochromatin protein 1β, and senescence-associated-β-galactosidase activity. Senescent hepatocytes were characterised by gene expression arrays and quantitative polymerase chain reaction (qPCR), and conditioned media was used in proteomic analyses, a human chemokine protein array, and cell migration assays to characterise the composition and function of the hepatocyte SASP. RESULTS: Senescent hepatocytes induced classical markers of senescence (p21, heterochromatin protein 1β, and senescence-associated- β-galactosidase activity); and downregulated the proliferation marker, Ki67. Hepatocyte senescence induced a 4.6-fold increase in total secreted protein (P = 0.06) without major alterations in the protein profile. Senescence-induced genes were identified by microarray (Benjamini Hochbergcorrected P < 0.05);, consistent with the increase in secreted protein, gene ontology analysis revealed a significant enrichment of secreted proteins among inducible genes. The hepatocyte SASP included characteristic factors such as interleukin (IL)-8 and IL-6, as well as novel components such as SAA4, IL-32 and Fibrinogen, which were validated by qPCR and/ or chemokine protein array. Senescent hepatocyteconditioned medium elicited migration of inflammatory (granulocyte-macrophage colony stimulating factor, GM-CSF-derived), but not non-inflammatory (CSF-1-derived) human macrophages (P = 0.022), which could contribute to a pro-inflammatory microenvironment in vivo , or facilitate the clearance of senescent cells. CONCLUSION: Our novel model of hepatocyte senescence provides insights into mechanisms by which senescent hepatocytes may promote chronic liver disease pathogenesis.	en_US
dc.relation.ispartof	World Journal of Gastroenterology	en_US
dc.relation.isbasedon	10.3748/wjg.v20.i47.17851	en_US
dc.subject.classification	Gastroenterology & Hepatology	en_US
dc.subject.mesh	Macrophages	en_US
dc.subject.mesh	Hepatocytes	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Hydrogen Peroxide	en_US
dc.subject.mesh	Inflammation Mediators	en_US
dc.subject.mesh	Biological Markers	en_US
dc.subject.mesh	Cytokines	en_US
dc.subject.mesh	Culture Media, Conditioned	en_US
dc.subject.mesh	Coculture Techniques	en_US
dc.subject.mesh	Cell Aging	en_US
dc.subject.mesh	Paracrine Communication	en_US
dc.subject.mesh	Chemotaxis	en_US
dc.subject.mesh	Gene Expression Regulation	en_US
dc.subject.mesh	Oxidative Stress	en_US
dc.subject.mesh	Dose-Response Relationship, Drug	en_US
dc.subject.mesh	Phenotype	en_US
dc.subject.mesh	Hep G2 Cells	en_US
dc.subject.mesh	Transcriptome	en_US
dc.subject.mesh	Cell Cycle Checkpoints	en_US
dc.subject.mesh	Biomarkers	en_US
dc.subject.mesh	Cellular Senescence	en_US
dc.title	Senescent human hepatocytes express a unique secretory phenotype and promote macrophage migration	en_US
dc.type	Journal Article
utslib.citation.volume	47	en_US
utslib.citation.volume	20	en_US
utslib.for	1103 Clinical Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access
pubs.issue	47	en_US
pubs.publication-status	Published	en_US
pubs.volume	20	en_US

Abstract:

© 2014 Baishideng Publishing Group Inc. All rights reserved. AIM: To develop a model of stress-induced senescence to study the hepatocyte senescence associated secretory phenotype (SASP). METHODS: Hydrogen peroxide treatment was used to induce senescence in the human HepG2 hepatocyte cell line. Senescence was confirmed by cytochemical staining for a panel of markers including Ki67, p21, heterochromatin protein 1β, and senescence-associated-β-galactosidase activity. Senescent hepatocytes were characterised by gene expression arrays and quantitative polymerase chain reaction (qPCR), and conditioned media was used in proteomic analyses, a human chemokine protein array, and cell migration assays to characterise the composition and function of the hepatocyte SASP. RESULTS: Senescent hepatocytes induced classical markers of senescence (p21, heterochromatin protein 1β, and senescence-associated- β-galactosidase activity); and downregulated the proliferation marker, Ki67. Hepatocyte senescence induced a 4.6-fold increase in total secreted protein (P = 0.06) without major alterations in the protein profile. Senescence-induced genes were identified by microarray (Benjamini Hochbergcorrected P < 0.05);, consistent with the increase in secreted protein, gene ontology analysis revealed a significant enrichment of secreted proteins among inducible genes. The hepatocyte SASP included characteristic factors such as interleukin (IL)-8 and IL-6, as well as novel components such as SAA4, IL-32 and Fibrinogen, which were validated by qPCR and/ or chemokine protein array. Senescent hepatocyteconditioned medium elicited migration of inflammatory (granulocyte-macrophage colony stimulating factor, GM-CSF-derived), but not non-inflammatory (CSF-1-derived) human macrophages (P = 0.022), which could contribute to a pro-inflammatory microenvironment in vivo , or facilitate the clearance of senescent cells. CONCLUSION: Our novel model of hepatocyte senescence provides insights into mechanisms by which senescent hepatocytes may promote chronic liver disease pathogenesis.

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http://hdl.handle.net/10453/118241