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Journal of Virology, September 2004, p. 9041-9050, Vol. 78, No. 17
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.17.9041-9050.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Global Effects of Human Papillomavirus Type 18 E6/E7 in an Organotypic Keratinocyte Culture System{dagger}

Peggy A. Garner-Hamrick,1 J. M. Fostel,1 Wei-Ming Chien,2 N. Sanjib Banerjee,2 Louise T. Chow,2 Thomas R. Broker,2 and Chris Fisher1*

Genomics-ID, Pharmacia Corp., Kalamazoo, Michigan,1 Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama2

Received 21 January 2004/ Accepted 18 April 2004

The effects of human papillomavirus type 18 (HPV-18) E6 and E7 proteins on global patterns of host gene expression in primary human keratinocytes grown in organotypic raft culture system were assessed. Primary human keratinocytes were infected with retroviruses that express the wild-type HPV-18 E6 and E7 genes from the native differentiation-dependent HPV enhancer-promoter. Total RNA was isolated from raft cultures and used to generate probes for querying Affymetrix U95A microarrays, which contain >12,500 human gene sequences. Quadruplicate arrays of each E6/E7-transduced and empty vector-transduced samples were analyzed by 16 pairwise comparisons. Transcripts altered in ≥12 comparisons were selected for further analysis. With this approach, HPV-18 E6/E7 expression significantly altered the expression of 1,381 genes. A large increase in transcripts associated with DNA and RNA metabolism was observed, with major increases noted for transcription factors, splicing factors, and DNA replication elements, among others. Multiple genes associated with protein translation were downregulated. In addition, major alterations were found in transcripts associated with the cell cycle and cell differentiation. Our study provides a systematic description of transcript changes brought about by HPV-18 E6/E7 in a physiologically relevant model and should furnish a solid source of information to guide future studies.

* Corresponding author. Mailing address: Director of Biology, NanoVir, LLC, and Interim CEO, Biosciences Research and Commercialization Center, Western Michigan University, 4717 Campus Dr., Suite 1300, Kalamazoo, MI 49008. Phone: (616) 833-1472. Fax: (616) 833-0992. E-mail: chris.fisher{at}nanovirpharm.com.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org.

Journal of Virology, September 2004, p. 9041-9050, Vol. 78, No. 17
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.17.9041-9050.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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