Analysis of Asbestos-Induced Gene Expression Changes in Bronchiolar Epithelial Cells Using Laser Capture Microdissection and Quantitative Reverse Transcriptase-Polymerase Chain Reaction

  • Christopher B. Manning
  • Brooke T. Mossman
  • Douglas J. Taatjes
Part of the Methods in Molecular Biology™ book series (MIMB, volume 319)


Laser capture microdissection (LCM) enables the removal of discrete microstructures or cell types from properly prepared histological sections. Extraction of RNA from microdissected tissue followed by quantitative reverse transcriptase-polymerase chain (QRT-PCR) reaction permits the analysis of cell-type or microstructure-specific gene expression changes that occur in response to various stimuli in the environment. In our lab, the combination of LCM and QRT-PCR has proven very useful in the determination of the in vivo gene expression changes that occur in bronchiolar epithelium in response to inhalation of crocidolite asbestos. A detailed description of the preparation of cDNA from bronchiolar epithelial cells obtained by LCM is described in this work.

Key Words

Asbestos bronchiolar epithelium environmental pathology laser capture microdissection lung disease microgenomics quantitative reverse transcriptase-polymerase chain reaction 


  1. 1.
    Emmert-Buck, M. R., Bonner, R. F., Smith, P. D., et al. (1996) Laser capture microdissection. Science 274, 998–1001.PubMedCrossRefGoogle Scholar
  2. 2.
    Taatjes, D. J., Palmer, C. J., Pantano, C., Hoffmann, S.B., Cummins, A., and Mossman, B.T. (2001) Laser-based microscopic approaches: application to cell signaling in environmental lung disease. Biotechniques 31, 880–894.PubMedGoogle Scholar
  3. 3.
    Manning, C. B., Cummins, A. B., Jung, M. W., et al. (2002) A mutant epidermal growth factor receptor targeted to lung epithelium inhibits asbestos-induced proliferation and proto-oncogene expression. Cancer Res. 62, 4169–4175.PubMedGoogle Scholar
  4. 4.
    PicoPure RNA Isolation Kit User Guide Version B, Arcturus Engineering, Mountain View, CA.Google Scholar
  5. 5.
    RQ1 RNase-Free DNase Technical Bulletin TB518, Promega Corporation.Google Scholar
  6. 6.
    Reverse Transcription System Technical Bulletin TB099, Promega Corporation.Google Scholar

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Christopher B. Manning
    • 1
  • Brooke T. Mossman
    • 1
  • Douglas J. Taatjes
    • 2
  1. 1.Department of PathologyUniversity of VermontBurlington
  2. 2.Department of Pathology, and Microscopy Imaging CenterUniversity of VermontBurlington

Personalised recommendations