Novel in vitro assays for the characterization of EMT in tumourigenesis

V. Koo, A. El Mekabaty, Peter Hamilton, Perry Maxwell, Osama Sharaf Eldin, J. Diamond, J. Watson, Kathleen Williamson

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background Two novel assays quantifying Epithelial to Mesenchymal Transition (EMT) were compared to traditional motility and migration assays. TGF-ß1 treatment of AY-27 rat bladder cancer cells acted as a model of EMT in tumourigenesis. Methods AY-27 rat bladder cancer cells incubated with 3ng/ml TGF-ß1 or control media for 24 or 48h were assessed using novel and traditional assays. The Spindle Index, a novel measure of spindle phenotype, was derived from the ratio of maximum length to maximum width of cells. The area covered by cells which migrated from a fixed coverslip towards supplemented agarose was measured in a novel chemoattractant assay. Motility, migration and immunoreactivity for E-cadherin, Vimentin and cytokeratin were assessed. Results TGF-ß1 treated cells had increased “spindle” phenotype together with decreased E-cadherin, decreased Cytokeratin-18 and increased Vimentin immunoreactivity. After 48h, the mean Spindle Index of TGF-ß1 treated cells was significantly higher than Mock (p=0.02 Bonferroni test) and there were significant differences in migration across treatment groups measured using the novel chemoattractant assay (p = 0.02, Chi-Square). TGF-ß1 significantly increased matrigel invasion. Conclusion The Spindle Index and the novel chemoattractant assay are valuable adjunctive assays for objective characterization of EMT changes during tumourigenesis.
Original languageEnglish
Pages (from-to)67-76
Number of pages10
JournalAnalytical Cellular Pathology
Volume32
Issue number1-2
DOIs
Publication statusPublished - 2010

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Medicine
  • Oncology

Fingerprint Dive into the research topics of 'Novel in vitro assays for the characterization of EMT in tumourigenesis'. Together they form a unique fingerprint.

  • Cite this

    Koo, V., El Mekabaty, A., Hamilton, P., Maxwell, P., Sharaf Eldin, O., Diamond, J., Watson, J., & Williamson, K. (2010). Novel in vitro assays for the characterization of EMT in tumourigenesis. Analytical Cellular Pathology, 32(1-2), 67-76. https://doi.org/10.3233/CLO-2009-0501