Document Detail


Subcellular localization and quantitation of the human papillomavirus type 16 E6 oncoprotein through immunocytochemistry detection.
MedLine Citation:
PMID:  23084288     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Human papillomavirus (HPV) type 16 E6 is a viral oncoprotein essential for host cell transformation. Due to its role in HPV-induced cancers of the genital, head and neck epithelia, reliable protein-level determination of E6 expression would be an invaluable diagnostic tool. Immunocytochemical detection and subcellular localization of HPV16 E6 has been demonstrated with varying success and a comprehensive review of techniques is lacking. To address these issues, we used established monoclonal antibodies and optimized a standard immunocytochemical method for E6 protein detection inside the HPV16 positive cell lines, SiHa and CaSki. E6 oncoprotein was detected primarily in the nucleus. We also refined quantitative analysis with a software to objectively differentiate between HPV16 positive and negative cells. Our analysis was also able to differentiate expression differences between SiHa and CaSki on par with RT-qPCR. Thus, we provide a long-needed, robust protocol for antibody-mediated detection of the HPV16 E6 oncoprotein inside cultured cells.
Authors:
Robert Jackson; Melissa Togtema; Ingeborg Zehbe
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-17
Journal Detail:
Title:  Virology     Volume:  -     ISSN:  1096-0341     ISO Abbreviation:  Virology     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0110674     Medline TA:  Virology     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
Affiliation:
Thunder Bay Regional Research Institute, Probe Development & Biomarker Exploration, 980 Oliver Road, Thunder Bay, Canada ON P7B 6V4; Department of Biology, Lakehead University, Thunder Bay, Canada ON P7B 5E1. Electronic address: rjackso1@lakeheadu.ca.
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