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Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment.

Publication ,  Journal Article
Buckanovich, RJ; Sasaroli, D; O'brien-Jenkins, A; Botbyl, J; Conejo-Garcia, JR; Benencia, F; Liotta, LA; Gimotty, PA; Coukos, G
Published in: Cancer Biol Ther
June 2006

Expression profiling using microarrays has become an essential tool for interrogating tumor biology. However, profiling of whole tumor RNA reflects both tumor and host cells, making it difficult to dissect molecular events within specific cellular compartments in the tumor microenvironment. We developed and optimized a simple, rapid technique combining immunohistochemistry and laser-capture microdissection (immuno-LCM) to purify specific cell populations from the tumor microenvironment followed by RNA isolation and amplification for microarray analysis. Using this methodology, we were able to elucidate the in situ expression profile of pure tumor cells and tumor endothelial cells from ovarian tumors with brisk immune infiltrates. This technique not only increased the specificity of profiling isolated cell populations, eliminating genes expressed by surrounding cells, but also increased the sensitivity of analysis, allowing for the detection of low expression genes that were not detected in whole tumor arrays. Pathway analysis of tumor cells in situ identified distinct activation of signaling pathways converging on NF-kappaB, as compared to pathways identified in cultured tumor cell lines, which were primarily metabolic. Profiling of tumor vascular cells revealed most known panendothelial and tumor endothelial-specific markers, and unveiled genes specific to the myeloid-monocytic lineage. We propose that immuno-LCM coupled with transcriptional profiling is a convenient tool for dissecting molecular and cellular events in complex biological systems such as the tumor microenvironment.

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Published In

Cancer Biol Ther

DOI

ISSN

1538-4047

Publication Date

June 2006

Volume

5

Issue

6

Start / End Page

635 / 642

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Polymerase Chain Reaction
  • Ovarian Neoplasms
  • Oncology & Carcinogenesis
  • Neoplasms
  • Microdissection
  • Lasers
  • Immunohistochemistry
  • Humans
  • Gene Expression Profiling
 

Citation

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Buckanovich, R. J., Sasaroli, D., O’brien-Jenkins, A., Botbyl, J., Conejo-Garcia, J. R., Benencia, F., … Coukos, G. (2006). Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment. Cancer Biol Ther, 5(6), 635–642. https://doi.org/10.4161/cbt.5.6.2676
Buckanovich, Ronald J., Dimitra Sasaroli, Ann O’brien-Jenkins, Jeffrey Botbyl, Josè R. Conejo-Garcia, Fabian Benencia, Lance A. Liotta, Phyllis A. Gimotty, and George Coukos. “Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment.Cancer Biol Ther 5, no. 6 (June 2006): 635–42. https://doi.org/10.4161/cbt.5.6.2676.
Buckanovich RJ, Sasaroli D, O’brien-Jenkins A, Botbyl J, Conejo-Garcia JR, Benencia F, et al. Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment. Cancer Biol Ther. 2006 Jun;5(6):635–42.
Buckanovich, Ronald J., et al. “Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment.Cancer Biol Ther, vol. 5, no. 6, June 2006, pp. 635–42. Pubmed, doi:10.4161/cbt.5.6.2676.
Buckanovich RJ, Sasaroli D, O’brien-Jenkins A, Botbyl J, Conejo-Garcia JR, Benencia F, Liotta LA, Gimotty PA, Coukos G. Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment. Cancer Biol Ther. 2006 Jun;5(6):635–642.

Published In

Cancer Biol Ther

DOI

ISSN

1538-4047

Publication Date

June 2006

Volume

5

Issue

6

Start / End Page

635 / 642

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Polymerase Chain Reaction
  • Ovarian Neoplasms
  • Oncology & Carcinogenesis
  • Neoplasms
  • Microdissection
  • Lasers
  • Immunohistochemistry
  • Humans
  • Gene Expression Profiling