Genome-wide gain-of-function screen for genes that induce epithelial-to-mesenchymal transition in breast cancer

Oncotarget. 2016 Sep 20;7(38):61000-61020. doi: 10.18632/oncotarget.11314.

Abstract

Epithelial to mesenchymal transition (EMT) is a developmental program that has been implicated in progression, metastasis and therapeutic resistance of some carcinomas. To identify genes whose overexpression drives EMT, we screened a lentiviral expression library of 17000 human open reading frames (ORFs) using high-content imaging to quantitate cytoplasmic vimentin. Hits capable of increasing vimentin in the mammary carcinoma-derived cell line MDA-MB-468 were confirmed in the non-tumorigenic breast-epithelial cell line MCF10A. When overexpressed in this model, they increased the rate of cell invasion through Matrigel™, induced mesenchymal marker expression and reduced expression of the epithelial marker E-cadherin. In gene-expression datasets derived from breast cancer patients, the expression of several novel genes correlated with expression of known EMT marker genes, indicating their in vivo relevance. As EMT-associated properties are thought to contribute in several ways to cancer progression, genes identified in this study may represent novel targets for anti-cancer therapy.

Keywords: breast cancer; epithelial-mesenchymal transition; high-content-screening; lentiviral vectors; vimentin.

MeSH terms

  • Antigens, CD
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology*
  • Cadherins / metabolism
  • Cell Line, Tumor
  • Disease Progression
  • Epithelial Cells / metabolism
  • Epithelial-Mesenchymal Transition*
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic
  • Genome, Human*
  • Genome-Wide Association Study
  • Humans
  • Lentivirus / metabolism
  • Open Reading Frames
  • Plasmids / metabolism
  • Vimentin / metabolism

Substances

  • Antigens, CD
  • CDH1 protein, human
  • Cadherins
  • Vimentin