Liquid-based Pap Smears as a Source of RNA for Gene Expression Analysis

Ellen F. Hildebrandt, Jeffrey R. Lee, John H. Crosby, Daron G. Ferris, Mark G. Anderson

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The Papanicolaou smear has contributed to a decrease in cervical cancer rates in populations that receive regular screening. However, treatment of women with mildly abnormal cells is problematic because the majority of these women do not develop neoplasia. Thus, new techniques for identification of truly precancerous cells are needed. Characterization of cellular gene expression patterns is now possible through microarray techniques that survey the expression of large numbers of genes simultaneously. Here we have assessed the feasibility of combining new microscopic and molecular technologies to determine gene expression patterns in cervical intraepithelial neoplasia grade 3 cells recovered from liquid cytology-based Papanicolaou smear slides. Laser capture microdissection was used to retrieve cervical cells from ThinPrep prepared slides. The quality of RNA recovered from these cells proved suitable for reverse transcription polymerase chain reaction and for T7 RNA polymerase-based linear amplification of messenger RNA. We developed an optimized RNA amplification protocol that permitted microarray gene expression profiling in samples of as few as 20 cervical cells. This approach combining laser capture microdissection, linear RNA amplification, and microarray gene expression analysis will enable comparison of gene expression patterns between cytologically abnormal and normal cells taken from a single slide and may assist in the differential diagnosis of histologically difficult cases.

Original languageEnglish (US)
Pages (from-to)345-351
Number of pages7
JournalApplied Immunohistochemistry and Molecular Morphology
Volume11
Issue number4
DOIs
StatePublished - Dec 2003

Keywords

  • Cervical cancer
  • Gene Chip
  • Papanicolaou smear
  • RNA amplification

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Medical Laboratory Technology

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