| Hepatocyte growth factor stimulates adenoviral-mediated gene transfer across the apical membrane of epithelial cells. | |
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MedLine Citation:
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PMID: 15170733 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: The apical surface of polarized epithelial cells is relatively resistant to gene delivery by various agents including adenoviral vectors. Hepatocyte growth factor (HGF) dedifferentiates previously well-polarized Madin-Darby canine kidney (MDCK) cell monolayers by altering cell-surface polarity and inhibiting tight junction function. METHODS: We used an in vitro model of polarized MDCK cells grown on permeable supports to examine the effects of HGF pretreatment on adenoviral (Ad)-mediated gene delivery through the apical surface of epithelial cell monolayers. RESULTS: HGF pretreatment of MDCK cell monolayers for 72 h increased Ad-mediated gene transfer and expression of enhanced green fluorescent protein (EGFP) and luciferase in a dose-dependent fashion. Time-course analysis of HGF-induced stimulation of Ad-mediated gene transfer was seen after 24 h and increased further with pretreatment periods extending to 72 h. HGF pretreatment increased Ad-mediated gene transfer at varying multiplicity of infection (MOI; ranging from 0.2-2000). PCR analysis for adenoviral DNA in control and HGF-pretreated MDCK cells suggested increased entry of viral constructs into HGF-pretreated MDCK cell monolayers. HGF-induced alterations in cell polarity are reversible upon removal of HGF. CONCLUSIONS: These data demonstrate that HGF pretreatment of MDCK cells increases the sensitivity of the cells to Ad-mediated gene delivery. The mechanism by which this occurs appears to be through increased entry of adenovirus into epithelial cells. These data provide evidence that biological agents that transiently alter epithelial cell polarity and tight junction function can be used to augment Ad-mediated gene delivery into epithelial cells from the apical surface. |
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Authors:
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Jason K Burrus; William D Matheson; Jeong S Hong; Eric J Sorscher; Daniel F Balkovetz |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: The journal of gene medicine Volume: 6 ISSN: 1099-498X ISO Abbreviation: J Gene Med Publication Date: 2004 Jun |
Date Detail:
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Created Date: 2004-06-01 Completed Date: 2005-01-28 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 9815764 Medline TA: J Gene Med Country: England |
Other Details:
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Languages: eng Pagination: 624-30 Citation Subset: IM |
Copyright Information:
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Copyright 2004 John Wiley & Sons, Ltd. |
Affiliation:
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Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35294, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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genetics* Cell Compartmentation Cell Membrane / drug effects, physiology* Cell Polarity / drug effects Cells, Cultured Dose-Response Relationship, Drug Epithelial Cells / drug effects, physiology* Gene Transfer Techniques* Green Fluorescent Proteins / genetics, metabolism Hepatocyte Growth Factor / pharmacology* Kidney / cytology, drug effects Luciferases / genetics Polymerase Chain Reaction / methods Time Factors |
| Grant Support | |
ID/Acronym/Agency:
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P30 DK54781/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
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147336-22-9/Green Fluorescent Proteins; 67256-21-7/Hepatocyte Growth Factor; EC 1.13.12.-/Luciferases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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