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Detection and cellular localization of heparin-binding epidermal growth factor-like growth factor mRNA and protein in human atherosclerotic tissue

Detection and cellular localization of heparin-binding epidermal growth factor-like growth factor mRNA and protein in human atherosclerotic tissue
Detection and cellular localization of heparin-binding epidermal growth factor-like growth factor mRNA and protein in human atherosclerotic tissue
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the epidermal growth factor family which binds to and activates the epidermal growth factor (EGF) receptor. HB-EGF mRNA is expressed by monocytes and vascular smooth muscle cells (VSMC) in culture, and has been shown to be a potent VSMC mitogenin vitro. The aim of this study was to screen normal and human atherosclerotic arteries and SMC cultured from these arteries for expression of HB-EGF, and to determine its cellular localization in human lesions. Using the highly sensitive technique of reverse transcription polymerase chain reaction (RT-PCR), we screened biopsies taken from normal human vessel walls and atherosclerotic tissue, for expression of HB-EGF mRNA. Northern blotting and RT-PCR were employed to determine levels of HB-EGF gene expression in SMC, cultured from normal and atherosclerotic arteries. Cellular localization of mRNA and protein, within human atherosclerotic plaques, was assessed using in situ hybridization with 35S labelled riboprobes, and immunohistochemistry with polyclonal antibodies specific for human HB-EGF. HB-EGF mRNA was found to be expressed in human atherosclerotic lesions and in VSMC cultured from these lesions. Expression of HB-EGF could not be detected in quiescent aortic VSMC using Northern blotting, but was highly up-regulated in these cells after treatment with basic fibroblast growth factor (bFGF) for 24 h. Although HB-EGF mRNA was detected in all vascular tissue examined using RT-PCR, in situ hybridization and immunohistochemistry revealed expression of HB-EGF in small portions of diseased arteries only. Immunohistochemistry showed strong staining for macrophages in all areas of HB-EGF expression. No association of HB-EGF with SMC was observed in any of the specimens examined. In conclusion, HB-EGF, a potent mitogen for VSMC, is expressed by macrophages in human atherosclerotic arteries, suggesting a role for the growth factor in the development of atherosclerotic plaques.
1639-1648
Reape, Theresa J.
45b5f181-c18a-404c-9181-7f5127498830
Wilson, Victoria J.
3b6ae12b-7629-466a-bbad-599cfb2cc24d
Kanczler, Janos M.
eb8db9ff-a038-475f-9030-48eef2b0559c
Ward, Jeremy P.T.
9f274c65-78b3-4ded-aeb8-e25e0bea2794
Burnand, Kevin G.
fc70ca87-5825-4eac-9507-98ea7a02fae2
Thomas, Chris R..
42b8f1ab-0c57-47eb-8e7c-02f5abc4e4c4
Reape, Theresa J.
45b5f181-c18a-404c-9181-7f5127498830
Wilson, Victoria J.
3b6ae12b-7629-466a-bbad-599cfb2cc24d
Kanczler, Janos M.
eb8db9ff-a038-475f-9030-48eef2b0559c
Ward, Jeremy P.T.
9f274c65-78b3-4ded-aeb8-e25e0bea2794
Burnand, Kevin G.
fc70ca87-5825-4eac-9507-98ea7a02fae2
Thomas, Chris R..
42b8f1ab-0c57-47eb-8e7c-02f5abc4e4c4

Reape, Theresa J., Wilson, Victoria J., Kanczler, Janos M., Ward, Jeremy P.T., Burnand, Kevin G. and Thomas, Chris R.. (1997) Detection and cellular localization of heparin-binding epidermal growth factor-like growth factor mRNA and protein in human atherosclerotic tissue. Journal of Molecular and Cellular Cardiology, 29 (6), 1639-1648. (doi:10.1006/jmcc.1997.0399). (PMID:9220349)

Record type: Article

Abstract

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the epidermal growth factor family which binds to and activates the epidermal growth factor (EGF) receptor. HB-EGF mRNA is expressed by monocytes and vascular smooth muscle cells (VSMC) in culture, and has been shown to be a potent VSMC mitogenin vitro. The aim of this study was to screen normal and human atherosclerotic arteries and SMC cultured from these arteries for expression of HB-EGF, and to determine its cellular localization in human lesions. Using the highly sensitive technique of reverse transcription polymerase chain reaction (RT-PCR), we screened biopsies taken from normal human vessel walls and atherosclerotic tissue, for expression of HB-EGF mRNA. Northern blotting and RT-PCR were employed to determine levels of HB-EGF gene expression in SMC, cultured from normal and atherosclerotic arteries. Cellular localization of mRNA and protein, within human atherosclerotic plaques, was assessed using in situ hybridization with 35S labelled riboprobes, and immunohistochemistry with polyclonal antibodies specific for human HB-EGF. HB-EGF mRNA was found to be expressed in human atherosclerotic lesions and in VSMC cultured from these lesions. Expression of HB-EGF could not be detected in quiescent aortic VSMC using Northern blotting, but was highly up-regulated in these cells after treatment with basic fibroblast growth factor (bFGF) for 24 h. Although HB-EGF mRNA was detected in all vascular tissue examined using RT-PCR, in situ hybridization and immunohistochemistry revealed expression of HB-EGF in small portions of diseased arteries only. Immunohistochemistry showed strong staining for macrophages in all areas of HB-EGF expression. No association of HB-EGF with SMC was observed in any of the specimens examined. In conclusion, HB-EGF, a potent mitogen for VSMC, is expressed by macrophages in human atherosclerotic arteries, suggesting a role for the growth factor in the development of atherosclerotic plaques.

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Published date: June 1997

Identifiers

Local EPrints ID: 177641
URI: http://eprints.soton.ac.uk/id/eprint/177641
PURE UUID: 40507cf3-e567-4e80-b116-7acd8167c38b
ORCID for Janos M. Kanczler: ORCID iD orcid.org/0000-0001-7249-0414

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Date deposited: 18 Mar 2011 09:44
Last modified: 14 Mar 2024 02:50

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Contributors

Author: Theresa J. Reape
Author: Victoria J. Wilson
Author: Janos M. Kanczler ORCID iD
Author: Jeremy P.T. Ward
Author: Kevin G. Burnand
Author: Chris R.. Thomas

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