Effect of downregulation of ZEB1 on vimentin expression, tumour migration and tumourigenicity of melanoma B16F10 cells and CSCs

Cell Biol Int. 2014 Apr;38(4):452-61. doi: 10.1002/cbin.10223. Epub 2014 Feb 20.

Abstract

Zinc-finger E-box binding homeobox 1 (ZEB1) is a master regulator of epithelial-mesenchymal transition (EMT) and has been implicated in primary epithelial cancer biological processes, such as invasion and metastasis. However, the role of ZEB1 in progression of melanoma and cancer stem cells (CSCs) remains obscure. In this study, the recombinant plasmids of t3 shRNAs targeting mouse ZEB1 were constructed and transfected into melanoma B16F10 cells. The stable transfected cells were selected and the characteristics of ZEB1 downregulated B16F10 cells was assessed. The tumourigenicity of CD44(+) CD133(+) CSCs isolated from B16F10 cells stably transfected with the ZEB1-shRNA2 recombinant was also assessed. ZEB1-shRNAs B16F10 showed a lower expression of ZEB1 and vimentin, weaker migration, invasiveness, colony forming, and proliferation, and a lower tumourigenicity than the control cells. The tumourigenicity of the ZEB1-shRNA2 CD44(+) CD133(+) CSCs was also inhibited. In conclusion, ZEB1-shRNA2-mediated downregulation of ZEB1 expression in B16F10 cells and CSCs is involved in the inhibition of the EMT process. ZEB1 may be a potential target in melanoma targeted.

Keywords: RNA interference; cancer stem cells; epithelial-mesenchymal transition; melanoma; zinc finger E-box-binding protein 1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Movement
  • Cell Transformation, Neoplastic
  • Down-Regulation
  • Homeodomain Proteins / antagonists & inhibitors
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Kruppel-Like Transcription Factors / antagonists & inhibitors
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / metabolism*
  • Melanoma / metabolism
  • Melanoma / pathology
  • Mice
  • Mice, Inbred C57BL
  • Neoplastic Stem Cells / cytology
  • Neoplastic Stem Cells / metabolism*
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Vimentin / metabolism*
  • Zinc Finger E-box-Binding Homeobox 1

Substances

  • Homeodomain Proteins
  • Kruppel-Like Transcription Factors
  • RNA, Small Interfering
  • Vimentin
  • ZEB1 protein, mouse
  • Zinc Finger E-box-Binding Homeobox 1