Skip to main content

Purification of Diseased Cells from Barrett’s Esophagus and Related Lesions by Laser Capture Microdissection

  • Protocol
  • First Online:
Book cover Laser Capture Microdissection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 755))

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Devesa SS, Blot WJ, Fraumeni JF, Jr. (1998) Changing patterns in the incidence of esophageal and gastric carcinoma in the United States. Cancer. 83:2049–2053.

    Article  CAS  Google Scholar 

  2. Spechler SJ, Goyal RK. (1986) Barrett’s esophagus. N Engl J Med. 315:362–371.

    Article  CAS  Google Scholar 

  3. Blot WJ, McLaughlin JK. (1999) The changing epidemiology of esophageal cancer. Semin Oncol. 26:2–8.

    CAS  Google Scholar 

  4. Morales CP, Lee EL, Shay JW. (1998) In situ hybridization for the detection of telomerase RNA in the progression from Barrett’s esophagus to esophageal adenocarcinoma. Cancer. 83:652–659.

    Article  CAS  Google Scholar 

  5. Lord RV, Salonga D, Danenberg KD, et al. (2000) Telomerase reverse transcriptase expression is increased early in the Barrett’s metaplasia, dysplasia, adenocarcinoma sequence. J Gastrointest Surg. 4:135–142.

    Article  CAS  Google Scholar 

  6. Aisner DL, Wright WE, Shay JW. (2002) Telomerase regulation: not just flipping the switch. Curr Opin Genet Dev. 12:80–85.

    Article  CAS  Google Scholar 

  7. Cong YS, Wright WE, Shay JW. (2002) Human telomerase and its regulation. Microbiol Mol Biol Rev. 2002;66:407–425.

    Article  CAS  Google Scholar 

  8. Barclay JY, Morris A, Nwokolo CU. (2005) Telomerase, hTERT and splice variants in Barrett’s oesophagus and oesophageal adenocarcinoma. Eur J Gastroenterol Hepatol. 17:221–227.

    Article  CAS  Google Scholar 

  9. Yoneyama K, Aoyama N, Koizumi H, Tamai S. (1998) Telomerase activity in esophageal carcinoma and lesions unstained with Lugol’s solution. Nippon Rinsho. 56:1181–1185.

    CAS  Google Scholar 

  10. Bachor C, Bachor OA, Boukamp P. (1999) Telomerase is active in normal gastroin­testinal mucosa and not up-regulated in ­precancerous lesions. J Cancer Res Clin Oncol. 125:453–460.

    Article  CAS  Google Scholar 

  11. Going JJ, Fletcher-Monaghan AJ, Neilson L, et al. (2004) Zoning of mucosal phenotype, dysplasia, and telomerase activity measured by telomerase repeat assay protocol in Barrett’s esophagus. Neoplasia. 6:85–92.

    CAS  Google Scholar 

  12. Harle-Bachor C, Boukamp P. (1996) Telomerase activity in the regenerative basal layer of the ­epidermis inhuman skin and in immortal and carcinoma-derived skin keratinocytes. Proc Natl Acad Sci USA. 93:6476–6481.

    Article  CAS  Google Scholar 

  13. Emmert-Buck MR, Bonner RF, Smith PD, Chuaqui RF, Zhuang Z, Goldstein SR, Weiss RA, Liotta LA. (1996) Laser capture microdissection. Sci. 274(5289):998–1001.

    Article  CAS  Google Scholar 

  14. Collaud S, Wiedl T, Cattaneo E, Soltermann A, Hillinger S, Weder W, Arni S. (2010) Laser-capture microdissection impairs activity-based protein profiles for serine hydrolase in human lung ­adenocarcinoma. Biomol Tech. 21(1):25–28.

    Article  CAS  Google Scholar 

  15. Shammas MA, Qazi A, Batchu RB, et al. (2008) Telomere maintenance in laser capture microdissection-purified Barrett’s adenocarcinoma cells and effect of telomerase inhibition in vivo. Clin Cancer Res. 14:4971–4980.

    Article  CAS  Google Scholar 

  16. Huang Q, Yu C, Klein M, Fang J, Goyal RK. (2005) DNA index determination with Automated Cellular Imaging System (ACIS) in Barrett’s esophagus: comparison with CAS 200. BMC Clin Pathol. 5:7.

    Article  Google Scholar 

  17. Ohyama H, Zhang X, Kohno Y, et al. (2000) Laser capture microdissection-generated target sample for high-density oligonule‑tide array hybridization. Biotechniques. 29:530–536.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Masood A. Shammas .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Shammas, M.A., Rao, M.Y. (2011). Purification of Diseased Cells from Barrett’s Esophagus and Related Lesions by Laser Capture Microdissection. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 755. Humana Press. https://doi.org/10.1007/978-1-61779-163-5_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-163-5_14

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-162-8

  • Online ISBN: 978-1-61779-163-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics