Purification of diseased cells from Barrett's esophagus and related lesions by laser capture microdissection

Methods Mol Biol. 2011:755:181-7. doi: 10.1007/978-1-61779-163-5_14.

Abstract

Barrett's esophageal adenocarcinoma (BEAC) arises from Barrett's esophagus (BE), a premalignant lesion caused by acid reflux (heartburn). Although the cancer is uncommon, its incidence is rapidly rising in western countries. Like most other cancers, BEAC cells also have elevated telomerase activity which maintains telomere length and supports continued proliferation of these cells. It is not clear if telomerase is activated early at premalignant (BE) stage, because reports of telomerase activity in Barrett's and normal esophagi have been controversial. We have shown that detection of telomerase and telomeres becomes easier and much more reliable if purified BE cells are used instead of tissue specimens. This chapter, therefore, emphasizes the importance of laser capture microdissection and provides the method to purify Barrett's esophagus related cells, using this technique.

MeSH terms

  • Barrett Esophagus / metabolism
  • Barrett Esophagus / pathology*
  • Cell Separation / methods*
  • Cryopreservation
  • DNA / isolation & purification
  • Enzyme Assays
  • Epithelial Cells / pathology
  • Esophagus / pathology
  • Humans
  • Lasers*
  • Microdissection / methods*
  • Paraffin Embedding
  • RNA / isolation & purification
  • Telomerase / chemistry
  • Telomerase / metabolism

Substances

  • RNA
  • DNA
  • Telomerase