Application of laser capture microdissection and proteomics in colon cancer

Laura Catherine Lawrie, Stephanie Curran, H. L. McLeod, John E. Fothergill, Graeme Ian Murray

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

Aims-Laser capture microdissection is a recent development that enables the isolation of specific cell types for subsequent molecular analysis. This study describes a method for obtaining proteome information from laser capture microdissected tissue using colon cancer as a model.

Methods-Laser capture microdissection was performed on toluidine blue stained frozen sections of colon cancer. Tumour cells were selectively microdissected. Conditions were established for solubilising proteins from laser microdissected samples and these proteins were separated by two dimensional gel electrophoresis. Individual protein spots were cut from the gel, characterised by mass spectrometry, and identified by database searching. These results were compared with protein expression patterns and mass spectroscopic data obtained from bulk tumour samples run in parallel.

Results-Proteins could be recovered from laser capture microdissected tissue in a form suitable for two dimensional gel electrophoresis. The solubilised proteins retained their expected electrophoretic mobility in two dimensional gels as compared with bulk samples, and mass spectrometric analysis was also unaffected.

Conclusion-A method for performing two dimensional gel electrophoresis and mass spectrometry using laser capture microdissected tissue has been developed.

Original languageEnglish
Pages (from-to)253-258
Number of pages5
JournalMolecular Pathology
Volume54
Issue number4
DOIs
Publication statusPublished - 2001

Keywords

  • colon cancer
  • electrophoresis
  • proteomics
  • POLYACRYLAMIDE GELS
  • MESSENGER-RNA
  • PROTEINS
  • EXPRESSION

Cite this

Application of laser capture microdissection and proteomics in colon cancer. / Lawrie, Laura Catherine; Curran, Stephanie; McLeod, H. L.; Fothergill, John E.; Murray, Graeme Ian.

In: Molecular Pathology, Vol. 54, No. 4, 2001, p. 253-258.

Research output: Contribution to journalArticle

Lawrie, Laura Catherine ; Curran, Stephanie ; McLeod, H. L. ; Fothergill, John E. ; Murray, Graeme Ian. / Application of laser capture microdissection and proteomics in colon cancer. In: Molecular Pathology. 2001 ; Vol. 54, No. 4. pp. 253-258.
@article{1120c0857e4648b99797b1e971bfecf3,
title = "Application of laser capture microdissection and proteomics in colon cancer",
abstract = "Aims-Laser capture microdissection is a recent development that enables the isolation of specific cell types for subsequent molecular analysis. This study describes a method for obtaining proteome information from laser capture microdissected tissue using colon cancer as a model.Methods-Laser capture microdissection was performed on toluidine blue stained frozen sections of colon cancer. Tumour cells were selectively microdissected. Conditions were established for solubilising proteins from laser microdissected samples and these proteins were separated by two dimensional gel electrophoresis. Individual protein spots were cut from the gel, characterised by mass spectrometry, and identified by database searching. These results were compared with protein expression patterns and mass spectroscopic data obtained from bulk tumour samples run in parallel.Results-Proteins could be recovered from laser capture microdissected tissue in a form suitable for two dimensional gel electrophoresis. The solubilised proteins retained their expected electrophoretic mobility in two dimensional gels as compared with bulk samples, and mass spectrometric analysis was also unaffected.Conclusion-A method for performing two dimensional gel electrophoresis and mass spectrometry using laser capture microdissected tissue has been developed.",
keywords = "colon cancer, electrophoresis, proteomics, POLYACRYLAMIDE GELS, MESSENGER-RNA, PROTEINS, EXPRESSION",
author = "Lawrie, {Laura Catherine} and Stephanie Curran and McLeod, {H. L.} and Fothergill, {John E.} and Murray, {Graeme Ian}",
year = "2001",
doi = "10.1136/mp.54.4.253",
language = "English",
volume = "54",
pages = "253--258",
journal = "Molecular Pathology",
issn = "1366-8714",
publisher = "BMJ Publishing Group",
number = "4",

}

TY - JOUR

T1 - Application of laser capture microdissection and proteomics in colon cancer

AU - Lawrie, Laura Catherine

AU - Curran, Stephanie

AU - McLeod, H. L.

AU - Fothergill, John E.

AU - Murray, Graeme Ian

PY - 2001

Y1 - 2001

N2 - Aims-Laser capture microdissection is a recent development that enables the isolation of specific cell types for subsequent molecular analysis. This study describes a method for obtaining proteome information from laser capture microdissected tissue using colon cancer as a model.Methods-Laser capture microdissection was performed on toluidine blue stained frozen sections of colon cancer. Tumour cells were selectively microdissected. Conditions were established for solubilising proteins from laser microdissected samples and these proteins were separated by two dimensional gel electrophoresis. Individual protein spots were cut from the gel, characterised by mass spectrometry, and identified by database searching. These results were compared with protein expression patterns and mass spectroscopic data obtained from bulk tumour samples run in parallel.Results-Proteins could be recovered from laser capture microdissected tissue in a form suitable for two dimensional gel electrophoresis. The solubilised proteins retained their expected electrophoretic mobility in two dimensional gels as compared with bulk samples, and mass spectrometric analysis was also unaffected.Conclusion-A method for performing two dimensional gel electrophoresis and mass spectrometry using laser capture microdissected tissue has been developed.

AB - Aims-Laser capture microdissection is a recent development that enables the isolation of specific cell types for subsequent molecular analysis. This study describes a method for obtaining proteome information from laser capture microdissected tissue using colon cancer as a model.Methods-Laser capture microdissection was performed on toluidine blue stained frozen sections of colon cancer. Tumour cells were selectively microdissected. Conditions were established for solubilising proteins from laser microdissected samples and these proteins were separated by two dimensional gel electrophoresis. Individual protein spots were cut from the gel, characterised by mass spectrometry, and identified by database searching. These results were compared with protein expression patterns and mass spectroscopic data obtained from bulk tumour samples run in parallel.Results-Proteins could be recovered from laser capture microdissected tissue in a form suitable for two dimensional gel electrophoresis. The solubilised proteins retained their expected electrophoretic mobility in two dimensional gels as compared with bulk samples, and mass spectrometric analysis was also unaffected.Conclusion-A method for performing two dimensional gel electrophoresis and mass spectrometry using laser capture microdissected tissue has been developed.

KW - colon cancer

KW - electrophoresis

KW - proteomics

KW - POLYACRYLAMIDE GELS

KW - MESSENGER-RNA

KW - PROTEINS

KW - EXPRESSION

U2 - 10.1136/mp.54.4.253

DO - 10.1136/mp.54.4.253

M3 - Article

VL - 54

SP - 253

EP - 258

JO - Molecular Pathology

JF - Molecular Pathology

SN - 1366-8714

IS - 4

ER -