Analysis of gene expression in prostate cancer epithelial and interstitial stromal cells using laser capture microdissection.

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  • Author(s): Gregg JL;Gregg JL; Brown KE; Mintz EM; Piontkivska H; Fraizer GC
  • Source:
    BMC cancer [BMC Cancer] 2010 Apr 28; Vol. 10, pp. 165. Date of Electronic Publication: 2010 Apr 28.
  • Publication Type:
    Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 100967800 Publication Model: Electronic Cited Medium: Internet ISSN: 1471-2407 (Electronic) Linking ISSN: 14712407 NLM ISO Abbreviation: BMC Cancer Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : BioMed Central, [2001-
    • Subject Terms:
      Gene Expression Profiling*/methods ; Gene Expression Regulation, Neoplastic* ; Lasers*; Epithelial Cells/*metabolism ; Microdissection/*instrumentation ; Prostate/*metabolism ; Prostatic Neoplasms/*genetics ; Stromal Cells/*metabolism; Cell Line, Tumor ; Early Growth Response Protein 1/genetics ; Epithelial Cells/pathology ; Humans ; Immunohistochemistry ; Male ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Prostate/pathology ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; RNA, Messenger/metabolism ; Stromal Cells/pathology ; Tissue Array Analysis ; WT1 Proteins/genetics
    • Abstract:
      Background: The prostate gland represents a multifaceted system in which prostate epithelia and stroma have distinct physiological roles. To understand the interaction between stroma and glandular epithelia, it is essential to delineate the gene expression profiles of these two tissue types in prostate cancer. Most studies have compared tumor and normal samples by performing global expression analysis using a mixture of cell populations. This report presents the first study of prostate tumor tissue that examines patterns of differential expression between specific cell types using laser capture microdissection (LCM).
      Methods: LCM was used to isolate distinct cell-type populations and identify their gene expression differences using oligonucleotide microarrays. Ten differentially expressed genes were then analyzed in paired tumor and non-neoplastic prostate tissues by quantitative real-time PCR. Expression patterns of the transcription factors, WT1 and EGR1, were further compared in established prostate cell lines. WT1 protein expression was also examined in prostate tissue microarrays using immunohistochemistry.
      Results: The two-step method of laser capture and microarray analysis identified nearly 500 genes whose expression levels were significantly different in prostate epithelial versus stromal tissues. Several genes expressed in epithelial cells (WT1, GATA2, and FGFR-3) were more highly expressed in neoplastic than in non-neoplastic tissues; conversely several genes expressed in stromal cells (CCL5, CXCL13, IGF-1, FGF-2, and IGFBP3) were more highly expressed in non-neoplastic than in neoplastic tissues. Notably, EGR1 was also differentially expressed between epithelial and stromal tissues. Expression of WT1 and EGR1 in cell lines was consistent with these patterns of differential expression. Importantly, WT1 protein expression was demonstrated in tumor tissues and was absent in normal and benign tissues.
      Conclusions: The prostate represents a complex mix of cell types and there is a need to analyze distinct cell populations to better understand their potential interactions. In the present study, LCM and microarray analysis were used to identify novel gene expression patterns in prostate cell populations, including identification of WT1 expression in epithelial cells. The relevance of WT1 expression in prostate cancer was confirmed by analysis of tumor tissue and cell lines, suggesting a potential role for WT1 in prostate tumorigenesis.
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    • Grant Information:
      1CA331160 United States CA NCI NIH HHS
    • Accession Number:
      0 (EGR1 protein, human)
      0 (Early Growth Response Protein 1)
      0 (RNA, Messenger)
      0 (WT1 Proteins)
    • Publication Date:
      Date Created: 20100430 Date Completed: 20100816 Latest Revision: 20220225
    • Publication Date:
      20221213
    • Accession Number:
      PMC2876079
    • Accession Number:
      10.1186/1471-2407-10-165
    • Accession Number:
      20426842